An unique expressed sequence tag（EST）, which had the high similarity to the sequence of Arabidopsis thaliana LRR Cf-5 disease resistance protein-like, was found from an SSH cDNA library constructed using mRNA isolated from leaves of Chinese cabbage (Brassica campestris ssp. pekinensis) after Ecc (Erwinia carotovora subsp.carotovora) infection. RACE was used to extend the 5’and 3’unknown sequence of this EST. The cloned cDNA, named BcLRR, was 1286 bp including a complete open reading frame (ORF) and a complete 3’UTR with a putative poly (A) signal (GenBank Accession No.EU424347). The deduced polypeptide contained 327 amino acids and consisted of transmembrance domain on the N sequences and 8 extra-cytoplasmic LRR motifs, 7 of which had canonical LXXLXLXXN repeat units and one had non-canonical LXXLXXXXN unit. Amino acid sequence alignment analysis revealed that BcLRR displayed high homology of 96.6% with Arabidopsis Cf-5 and of 82.3％ with upland cotton LRR disease resistance protein-like GhLRR; The majority of their differences are confined to their N terminal signal peptide regions. Phylogenetic analysis further demonstrated that these three functionally unknown proteins were significantly distinct from other known resistance extra-cytoplasmic LRR proteins (Cf-2, Cf-4, Cf-5, and Cf-9 from tomato, Hs1pro-1 from sugarbeet and Xa21 from rice) and belonged to a new type of extra-cytoplasmic LRR protein. The complete ORF sequence of BcLRR was amplified and cloned into modified vector pGreen0029 under CaMV35S promoter, forming plant expressed vector pGreen0029-BcLRR, which was then transformed into Arabidopsis thaliana Col-0 via Agrobacterium GV3101/Soup. The results of PCR, Southern blotting and RT-PCR showed that BcLRR had been integrated into the Arabidopsis genome and expressed in the transgenic plants. BcLRR protein enhanced the resistance to Ecc BC1 in the transgentic Arabidopsisp plant.

Discovery of candidate site in virus-like particles(VLPs) of porcine parvovirus(PPV) for inserting foreign protein and the construction of the recombinant VLPs contained the ORF2 protein of porcine circovirus type 2(PCV2) ZHU Ling,GUO Wan-zhu**,JIANG Qing-rong,XU Zhi-wen,XIONG Ding-jie,ZHANG Bo,WANG Yin,ZHAO LING (Animal Biotechnology Center, Sichuan Agricultural University, YA’an,625014,China) Abstract:The plasmid pMD-VP2 was constructed by cloning the VP2 gene fragment gained by PCR from the PPV(SC-1)strain template into pMD18-T vector. The ORF2 gene fragment of PCV2(SC)strain was amplified using two pairs of primers ,each pair of which added identical restrict site at the 5’end of the forward and the downward (one pair added HindⅢ,the other added SacI),then the two PCR product were cloned into pMD18-T vector to construct pMD-ORF2.A and pMD-ORF2.B,respectively. Through digesting the two plasmid with the corresponding restrict endonuclease HindⅢ and SacI respectively, the expected fragments could be reclaim and were inserted into the restrict cloning sites, which were corresponding to the 1/3 position of the N-terminus and the C-terminus of the VP2 gene fragment in the pMD-VP2 ,and then the pPVP2－ORF2.A and pPVP2－ORF2.B were constructed. After verification, the correct directional cloning pPVP2－ORF2.A and pPVP2－ORF2.B were digested with restrict endonuclease KpnI、BamHI and ApaLI to get the expected gene fragments which contained VP2 and ORF2 gene, and then the two gene fragments were cloned into enkaryotic expression vector pEGFP-C1 to construct the two recombinant plasmid pEGFP.VO.A and pEGFP.VO.B. Employed fluorescence microscope and electron microscope to confirm the fusion gene expression after transfected the two recombinant plasmids into the COS-7 cell line with liposome, we found that VLPs form in the cell transfected the pEGFP.VO.A only but not in the group of pEGFP.VO.B. In animal, the BALB/C mice after been immunized the purified VLPs could produce comparatively high-level cytoimmunity and special humoral immunity against PPV and PCV2. The result indicated that the recombinant VLPs be composed of PPV-VP2 and PCV2-ORF2 is available in this study and revealed the 1/3 position of the N-terminus of the VP2 gene is suitable for inserting foreign protein to construct recombinant VLPs. Key Words: Porcine Parvovirus, Porcine Circovirus type 2, Virus-like particles, recombinant

A polygalacturonase inhibiting protein gene named FaPGIP was cloned from strawberry (Fragaria × ananassa) leaves through PCR.The FaPGIP gene from genomic DNA showed a single 168bp intron and the cDNA contained an open reading frame of 999bp, which encoded a polypeptide of 332 amino acid residues with a molecular mass of 37.1 kD and a pI of 7.67, and a hydrophobic region of 24 amino acid residues in the N-terminal which was considered to be a signal peptide. The deduced amino acid sequence contained 5 potential N-glycosylation sites. The sequence homology comparison showed that the FaPGIP exhibits a high homology of the nucleotide sequences with other PGIP genes from the GenBank. The three-dimensional model looked like a horseshoe-shaped structure, and contained 12 α-helices and 21 β-sheets, and the center LRR structural domain was composed of 10 tandem LRRs motifs. The expression of FaPGIP was found in root, stem, leaf, flower and fruit of strawberry. The expression levels in different tissues from fruit, leaf, flower, root and stem ranking high to low, with the relative content of 0.2460, 0.1664, 1.3712, 0.8721and 3.4154, respectively.

Genetic polymorphisms of milk epithelial mucin(MUC1) and high molecular protein were assayed in Yorkshire sow by using electrophoresis, in order to investigate the biodiversity of sow based on milk-specific proteins. Three types of MUC1 were revealed in 42 milk samples tested, with molecular weight of 230, 217, 212 kD, respectively. The molecular weights of these proteins are higher than those of cattle, and form 5 phenotypes of MUC1. Four types of high molecular protein(HMWP) were revealed in sow milk, with molecular weight of 123, 109, 107, 101 kD, respectively, and form 5 phenotypes of HMWP. Expressions of MUC1 and HMWP in sow colostrum within 3 d of lactation were lower than in milk. Sow milk contained higher content of MUC1 than cattle and goat. Results of the present study demonstrate that both MUC1 and HMWP shows polymorphism in sow milk, but their gene heterozygosity and effective number of alleles were relatively low.

Maize is the most important agronomic crop for human and livestock consumption, and is extensively used in modern foods, medicine and chemistry industry. In this study, coding DNA sequences(CDS) of 7402 proteins in maize were analyzed by Codon W, calculated the content of G+C at three positions of codons, ENc(effective number of codon) of genes and frequency of synonym codon usage, and determined the “optimal codons”. The results showed that the content of G+C at the third position of codons was significantly higher than that of the first and second positions, and genes prefer to the codons with C or G at the synonymous position. In addition, 27 codons were determined as the “optimal codons”, which were ending with G or C. The results provide evidences for these fields, such as molecular modification of exogenous gene to increase the expression efficiency, improving accuracy of gene prediction and genome annotation, and so on.

According to the highly conservative region of Hormone-sensitive Lipase (HSL) gene sequences from bovine (Bos taurus), human(Homo sapiens) and mouse (Mus musculus) in the GenBank, the specific primers were designed to clone the CDS region of HSL gene in mammary gland of Xinong Saanen dairy goats using the RT-PCR method, the bioinformatic analyses of nucleotide sequence, the deduced amino acid sequence, and the prediction of protein structures of HSL gene were conducted. The results showed that the full length of CDS region of HSL gene was 2271 bp encoding 756 amino acids (GenBank accession NO. EU273879), homogeneities of CDS nucleotide sequence between goat and bovine, human and mouse were 96%, 86%, and 79%; that of amino acid sequence were 96%, 85%, and 83%, respectively, the predicted molecular weight and isoelectric point of HSL gene were 82583.7 Da and 6.23, the similar structure of CDS region of HSL gene indicated the possible functions of HSL gene in milk secretion.

The 289 DH lines derived from the cross BS20×Fu3 were used for mapping QTLs of photoperiod-temperature sensitive genic male sterility in this study. Field trials were conducted in Beijing and Fuyang, Anhui Province, during 2005~2006 cropping season. The fertility investigated included seed setting rate and seed setting spikelet rate. The fertility-related QTLs were mapped using SSR markers and bulked segregant analysis (BSA). The genetic map consisted of 128 SSR markers. Three SSR loci Xgwm294, Xgwm374 and Xgwm44 on chromosomes 2AL, 2BS and 7DS, respectively, showed linkage with the fertility genes in the BSA analysis of sterile DNA pool and fertile DNA pool. Using mixed-model composite interval mapping, 6 QTLs were detected on chromosomes 1AS, 2BS, 2DL, 6AL, 6BL and 7DS, accounting for 1.1~12.5% of the phenotypic variance across two environments. Significant interactions were found between the QTL on 7DS and those on 2BS, 6AL and 6BL. Finally, two important QTLs were detected on chromosomes 7DS and 2BS in the marker intervals of Xgwm44-Xcfd14 and Xgwm148-Xgwm374, explaining 7.2~12.5％ and 2.1~2.5％ of phenotypic variances, respectively.

According to the principle of hybrid production, a new approach to restore the fertility of engineered male sterility induced by Barnase gene was constructed using the Cre/lox site-specific recombination system, which was distinct from the Barnase/Barstar restoring approach. In this study, the pBinBarloxTABn plant expression vector with a TA29-Barnase gene expression cassette flanked by two directly oriented lox sites and Bar gene expression cassette was transformed into Wisconsin 38 by Agrobacterium tumefaciens-mediated transformation. The male sterile plants appeared anther thinness, unfull and poor pollen produced. The pollen was unnormal, crimpy, and lost the germination ability, no normally expanded fruits and seeds formated were observed after self-pollinated. However,the normally fruits and seeds were obtained after pollinated using pollens from pBinCre transgenic plant. The hybrids were produced by pollinated BN1 and BN6 male sterile plants using pollen from pBinCre transgenic plant, total 23 F1 progenies from BN1 ×C1 hybrid, 22 F1 progenies from BN6 ×C1 hybrid were analyzed the TA29-Barnase gene deletion by PCR method.The results indicated the TA29-Barnase gene expression cassette had been excised efficiently from the genome of F1 progenies those having both Cre and Bar genes in them, the TA29-Barnase gene deletion rate reached 100% for both the BN1 ×C1 and BN6 ×C1 cross combinations. Those progenies that TA29-Barnase gene deleted could flower and fruit normally ,indicated the male sterility had been restored.

A cis-acting genetic element aps (amplification promoting sequence) was successfully isolated from ribosomal DNA of tobacco variety named “CuiBi No.1”. The aps encoded a complete nucleotide sequence of 440 bp, including TATA TAAATG motifs, and was rich in A/T sequences. The cloned sequence has been deposited into GenBank with accession No. FJ516382. The regulatory expression function of aps was examined by introducing it into upstream of CaMV35S promoter with GUS as a reporter gene. Southern blot result revealed that transgenic rice plants, transformed with expression cassettes, carrying the GUS gene and CaMV35S promoter in combination of aps, contained much more multiple copies when compared to those transformants without aps. The semi-quantitative RT-PCR analysis indicated that the transformants with the insertion of aps had much higher transcription levels. GUS activity quantitative analysis showed that transformants in fusion of aps had higher GUS activities with 12.07-16.03 (GUS activity /nmol MU mg protein-1min-1). The activity values were remarkable higher (*P<0.05) when compared to transformants lacking aps which had 5.32-7.01 GUS activity/nmol MU mg protein-1 min-1. Our results suggested that aps was likely to elevate the transgenic expression by increasing both target gene copy number and transcription level in rice, similar to its effects previously reported in dicot tobacco and tomato. It also suggested that aps could perform a specific function as an enhancer of transcription in monocot rice, implicating its potential application in plant genetic engineering.

Aegliops tauschii，one of the wild relatives of Triticum, is an important genetic resource for wheat improvement . In this study, 78 accessions of Ae. tauschii were analyzed using 211 pairs of wheat SSR primers .Our data showed that D genome of Ae. tauschii had greatest genetic similarity with D genome of wheat, followed by wheat B genome, and the last A genome. Thirty-two polymorphic SSR primers were selected, and 308 alleles were detected in Ae. tauschii accessions with an average of 8.14 alleles per locus detected by a pair of polymorphic pairs. The primers of A genome detected 39 alleles with a mean of 5.57 per locus, primers of B genome 51 alleles with a mean of 6.38 per locus, and primers of D genome 39 alleles with a mean of 12.47 per locus, which indicated that primers of D genome detected great genetic diversity. The result concluded that D genome of Ae. tauschii had greatest genetic similarity with D genome of common wheat, and it also had some similarity with A and B genome of common wheat since the primers of the two genomes were polymorphic in D genome of Ae. Tauschii.. UPGM A cluster analysis showed that tested materials could be classified into six clusters with similarity coefficiency at 0.77, and relationship of clusters was associated with geographical distribution of materials. genetic polymorphism were detected by SSR primers of D genome. UPGM A cluster analysis showed that all test materials could be classified into six main groups at similarities co-efficiency in 0.77. The cluster results were associated with its geographical distribution.

17 samples of Alternaria spp. which is the pathogen of wheat black point were collected from five different places in Henan, and their genomic DNAs were extracted by CTAB. The ribosomal DNA internal transcribed spacer (ITS) region of the genomic DNAs was amplified and sequenced, the homology of the 17 isolates was 99%-100%. The sequences were analyzed by phylogenesis with 9 species of Alternaria spp. from GenBank. The result showed that all the isolates had high homology The homology of the 6 small-spored species A. alternata、A. tenuissima、A. mali、A. gaisen、A. citri、A. arborescens is 98%-100%, and the other three large-spored species A. radicina、A. porri and A. solani were separated clearly. There are also some certain relative relationship between the mutation isolates of small-spored species and the large-spored ones. The rDNA ITS1-5.8S-ITS2 of all the isolates did not have the regionally and host characters but had high conservation with special sequences respectively, which can be the important evidence in the classification, identification, molecular marker and phylogenetic analyses of some certain species of epiphyte. The result also prove by molecular evidence that some similar small-spored species of Alternaria spp. are the pathogens of wheat black point in Henan.

According to the known mRNA sequence of bovine Oct4 gene in GenBank，a pairs of primers containing restriction sites of EcoR I and BglⅡ were designed, which were ultilized to amplify full sequence of ORF of bovine Oct4 gene. Total RNA was obtained from the primodial genital ridges of bovine fetus. The full-length of ORF of bovine Oct4 gene was 1,083 bp, which was obtained by RT-PCR reaction. The Homology of sequence cloned Oct4 gene was 99.4% to the counterpart sequence in Genbank. The recombinant plasmid pMSCV-Oct4 was transfected into packaging PT67 cells by lipofectamine 2000, at the same time, pMSCVneo-IRES-GFP vector (pMIG) was transfected into packaging PT67 cells as a control, which was used to measure transfected efficiency by flow cytometry analysis. Subsequently, we established a cell line stable producing retrovirus particles by G418 selection for 2 weeks. Retroviral titer was determinated with plaque formation method in NIH3T3 cell line, which was not less than 8.83×107cfu/mL. The transfection efficiency of pMSCV-Oct4 in PT67 cell line was 53.1%. The results indicated that the full sequence of bovine Oct4 gene was successfully cloned, and constructed recombinant retroviral pMSCV-Oct4 expression vector. Moreover, we established a packaging cell line stable producing retroviruses of Oct4 gene. The study must be helpful to the generation of induced pluripotent stem cells from bovine somatic cells.

A target fragment which contain two CCGG sites was cloned from the first coding region of S-locus receptor kinase gene (SRK) of Brassica oleracea with typical self-incompatibility (SI). Then the method combining methylation-sensitive restriction endonucleases with PCR (MS-RE-PCR) was used to analysis primarily the methylation of the specific DNA fragment of SRK gene coding region, that is, genomic DNA (gDNA) extracted from stigma papilla and anther and their PCR amplification products were digested and electrophored interlacedly and combindly. The restriction endonucleases were MspⅠand HpaⅡ，whose methylation-sensitivity were different. When using the same specific primer, the target bands could be got by using gDNA as template, and all target bands could also be digested completely by MspⅠ/HpaⅡ producing the expected smaller bands, but there was no specific PCR amplification products by using gDNA digested by MspⅠ/HpaⅡ as template. These results indicated that SRK gene in anther of SI B. oleracea may be not blocked by DNA methylation.

In the present study, the effect of different concentrations and different adding proposal of rhodiola polysaccharide on freezing boar spermatozoa was tested. Results: 1. rhodiola polysaccharide could provide better protective action for boar spermatozoa during cryopreservation, and especially anti-oxidation within the cooling period; 2. the effect of rhodiola polysaccharide added inⅠextender was better than that of added in Ⅱextender, and the effective concentration of rhodiola polysaccharide was 6 mg/L.

Five genes (cpcA, hox1, pcyA, cpcE and cpcF), involved in the biosynthesis of phycocyanin holo-α-subunit, were cloned from the genomic DNA from Spirulina maxima. These genes were digested by the corresponding restriction enzymes and sequentially ligated into the expression plasmid pETDuet-1 with two multiple cloning sites to construct the expression vector pETDuet-6. The recombinant vector pETDuet-6 was then transformed into E. coli BL21 (DE3) and ampicillin resistant transformant ZJGSU02 was selected. The fidelity of the recombinant vector was confirmed by DNA sequencing. After induction with isopropyl-β-D-thiogalactopyranoside (IPTG), the cells in the culture of the engineered strain exhibited a pronounced blue. The color change was not seen in the cells in the culture of the control strain. Visualization of the proteins on coomassie-stained SDS-PAGE gel showed about 21kDa distinct band. After the SDS-PAGE gel being soaked in zinc ion solution, a bright fluorescence band was visualized by UV illumination. Western blotting indicated the specific binding of the recombinant protein to 6×His-tag monoclonal antibody. The absorption spectrum of the recombinant phycocyanin holo-α-subunit had a λmax at 623nm, and the maximum fluorescence emission wavelength of it was at 645.8nm, which were in agreement with those of the natural phycocyanin holo-α-subunit from S. maxima. These results demonstrated that phycocyanin holo-α-subunit of S. maxima was successfully expressed in the heterologous E. coli. This study provided not only a new strategy for the expression of pigment-binding protein in the heterologous host, but also a reference for exploring the construction and expression of five or more heterologous protein genes using one vector and the study of the interaction among the genes.

Abstract: cDNA-AFLP technology and bioinformatics analysis was used to analyze gene expression difference at 10 consecutive periods bud from bud aborted to normal bud in Chinese cabbage, 192 differentially expressed sequences were obtained, in which 88 sequences were chosen randomly to sequence, 72 novel expressed sequence tags (ESTs) were obtained after sequencing. By bioinformatics analyzing, the ESTs were divided into seven types, such as energy and metabolism, membrane and transport, transcript and translation, amino acid synthesizing and processing, signal conduction, and Unclear classified et al, moreover, 5 ESTs were newly found in Chinese cabbage bud aborting and had been entered to GenBank, their entering number were GD185906、GD185907、GD185908、GD185909 and GD185910 respectively. The acquirement of ESTs related with Chinese cabbage aborted bud may be helpful to elucidate the mechanism of Chinese cabbage aborted bud, and will provide important resource for finding novel gene.

A pair of primers were designed and synthesized based on the sequences of the ARV and MDRV S3 genes,the S3 gene segment of the novel NP03 duck reovirus isolate could be specifically amplified by RT-PCR and be conducted sequence analysis. Virion RNA was isolated by treatment of aliquots of the virus with LS TRIAZOL. The cDNA of the S3 gene frame of J03 was amplified by reverse transcription-polymerase chain reaction ,cloned into pMDT-18 vector and sequenced,respectively.The results show that the cloned S3 gene includes the complete open reading frame (ORF).Evolution analysis demonstrated that the S3 nucleotide sequence from isolate J03 shares 60％～60.2％identity with chicken isolates, 61.9％identity with a turkey reovirus and 58.2％～62.7％identity with muscovy duck reoviruses,and 68.2％～69％,68.2％,63％～70.4％amino acid sequence identities with chicken isolates, a turkey reovirus, muscovy duck reoviruses,respectively.The results indicate that J03 is a novel duck reovirus is different from ARV and MDRV.

Azospirillum brasilense is a nitrogen fixing, plant growth promoting rhizobacterium, and it can interact with roots of several economically important cereals, such as maize, wheat, rice. NifA is a common transcriptional activator for all nitrogen fixation gene. In some diazotrophs, NifL controls negatively NifA activity. But there are not been found factors interaction with NifA in A. brasilense. The hybrid two component system protein Org35 was obtained by means of the yeast two-hybrid system with NifA as bait in our previous research. In this study, the interaction between Org35 and NifA was discussed. The results indicated the PAS domain was mediated in the interaction between Org35 and NifA.

A composite cross population was derived from the cross between (Yumian 1 × CRI 35) F1 and (Yumian 1 × Belshinuo) F1. A total of 6,565 primer pairs were used to screen the polymorphic primers between CRI 35, Yumian 1 and Belshinuo, and 92 polymorphic primer pairs were obtained. The polymorphic primer pairs were used to genotype the 172 individual plants of composite population and 96 loci were obtained. Four out of 92 primer pairs produced two loci. A genetic linkage map with 63 SSR loci and 19 linkage map was constructed, and the linkage map covered a whole length of 656 cM with an average interval of 10.41cM between two markers, accounting for 14.8% of the cotton genome. The yield traits of the 172 composite population F2 lines were used to detect QTL by multiple QTL mapping, and 7 QTL for five yield traits were identified, including one for boll number per plant (BN), two for boll weight (BW)，one for lint percentage (LP), one for seed cotton yield per plant (SC), and two for lint cotton yield per plant (LC). Seven QTL were mapped on four D-genome chromosomes, and four QTL were mapped on chromosome 19. It was found that the different parents have different QTL alleles of yield trait.

Construction of bifunctional fusion protein which can conjugate both human red blood cell and antibody against PRV.2E8 gene(ScFv gene against H antigen of human erythrocytes)and kgE gene (the major epitope domain of glycoprotein gE of pseudorabies virus) ,amplified from the recombinant plasmid PMD-2E8ScFv and PMD-gE,respectvely,were spliced by overlaped extension PCR and the assembled bifunctional molecule was named 2E8kgE.The 2E8kgE digested with BamHI and EcoRI was inserted into pET-Trx vector,to yield the recombinan vector pET-Trx-2E8kgE.Then the recombinan vector pET-Trx-2E8kgE was transformed into BL21plysS and protein expressions was induced with IPTG.SDS-PAGE and western-blot analyses showed that the fusion peotein was 60.1kDa and the protein was specific to antisera against PRV. Erythrocyte agglutination test results indicated that 2E8kgE fusion protein can conjugate both human red blood cell and antibody against PRV.

Abstract: Here we report a novel assay method of porcine reproductive and respiratory syndrome virus (PRRSV) termed reverse transcriptase loop-mediated isothermal amplification (RT-LAMP), using fluorescent reagent as the LAMP product judgment. With one set of special primers targeting sequence containing the M gene and N gene together, the LAMP-based assay can be completed within 30 min at 63℃. The detect limit of the assay is 1000 copies per reaction determined by using a recombined plasmid containing the target sequence, which is the same as RT-PCR assay. The specificities of the primers were confirmed using genomes of various viruses. In the fluorescent reagent test, 0.05mM calcein and 0.06mM manganese were confirmed to be appropriate in LAMP product judgment, and the result was the same as turbidity test. Furthermore, RT-LAMP assays were applied to the rapid detection of PRRSV in blood samples from 103 field piglets, 56 and 48 piglets, respectively, were positive by the RT-LAMP and RT-PCR. These results suggest that the RT-LAMP assay with fluorescent reagent provides a useful tool for the diagnosis of PRRSV infections in porcine rapidly.

PeaT1 is a protein elicitor from Alternaria tenuissima with a nascent polypeptide associated complex (NAC) domain. In this research, peaT1 was cloned by PCR and ligated to vector pET28a for expression. Then PeaT1 was purified and detected by synchrotron radiation circular dichroism to clarify the relations between heat and its secondary structures. The treatment temperature rose from 4℃, 25℃, 55℃ to 85℃, and then fell from 85℃, 55℃, 25℃ to 4℃. The proportions of α helix and turnel both decreased and then recoverd, the proportion of random coil increased and then recoverd, while the proportion of β sheet kept stable. Further analysis combinded with the structure information of PeaT1 indicated that the NAC domain which formed a homodimer played an essential role in heat resistance. This research illuminated the heat resistance mechanism of PeaT1 and laid the foundation for production and application of PeaT1 as a biological pesticide.

In the experiment, PCR-RFLP technique was used to detect the alleles distribution of S631N polymorphism site of myxo-virus resistance gene (Mx) in BAI chicken populations. Populations with different genotypes were constructed by selected mating method. After vaccine were injected the Newcastle Disease (ND) and Avian Influenza (AI) antibody titers of individuals with different genotypes were respectively measured at 35, 42, 49, 70 and 100 days of age using haemagglutination activity (HA) text and hemagglutination inhibition (HI) text. The body weight of 90 days, age at first laying and egg number at 72 weeks of individuals with different genotypes were also measured. The association between the polymoephism site and immune traits and production performance were analyzed. The resulted showed the allele frequencies at this locus was in Hardy-Weinberg equilibrium (P>0.05) in BAI populations, the frequencies of three different genotypes were 0.233(AA), 0.517(AG) and 0.250(GG). In tested days, the ND and AI antibody titers with different genotypes had the same increased and decreased laws, but individuals with AA genotypes had higher levels (P<0.05) than that of AG and GG genotypes in several tested days. The association analysis showed this polymorphism site had significant effect (P<0.05) on peak ND antibody titer of 49 days and AI antibody titer of 70 days. However, the body weight of 90 days, age at first laying and egg number at 72 weeks had no variation among individuals with different genotypes. The results identified the S631N polymorphism site of Mx gene as an important candidate mark for improving antiviral activity.

C. jejuni isolated from different sources (n=113) were subtyped by PCR-RAPD. The PCR products showed 15 different banding profiles, In this study, V7、V9、V10、V11、V12、V13、V14、V15, inhabited 81.42％ profile numberV10、V11、V12 and V14 were predominant in the isolates.Which were 11.50％、12.39％、13.27％ and 10.62%.But C1-C5、V6、V8 were lower than 1%.Besides, isolated from different sources with V7、V10、V11、V12、V14.It showed that isolates from different sources with a high degree of cross-distribution. Of the 113 C. jejuni isolates tested, the average positive rates of the adhesion related gene cadF, racR were 92.45% and 38.69%, the flagellar gene flaA was 73.58%, but the average rates of the virulence-associated genes cdtA、cdtB、cdtC、wlaN and virB11 were 71.70%、52.83%、96.23%、12.26% and 1.89%. Average rates of the heat shock protein and transporter associated protein gene dnaJ、ceuE were0.94%和65.09%，Average rates of gene CiaB、pldA were 39.62% and 9.43%，58.82% strains carried over 6 virulence-associated genes.

In recent years, the heart fatty acid-binding protein (H-FABP) gene was regarded as one candidate gene that affected intra-muscle fat (IMF) deposition, which was generally thought to be a major factor that influences meat quality of livestock. Three of variations (SNP or in/del) in this gene were genotyped in a F2 full sib of Xinhua ×White Recessive Rock reference population, with the purpose of discovering their associations with chicken fatty traits. The expression patterns within different tissues, day-old, genotypes of H-FABP gene were further studied in this study. The results showed that 1) the G+926A polymorphism was highly significantly associated with fat content of breast muscle (P<0.01); 2) the 14 bp in/del was highly significantly associated with body weight at the age of 70 days and 84 days (P<0.01), whereas no associations of this polymorphism with fat traits were observed at significant level; 3) the C+2851T polymorphism was highly significantly associated with fat content of leg muscle (P<0.01). The results of qRT-PCR and Haplotype were consistent with association study, this further suggested that H-FABP gene may be a candidate gene for chicken meat quality traits, such as IMF.

Abstract：Growth differentiation factor9(GDF9)is one of the members of transforming growth factorβ(TGFβ)，which is essential for growth and differentiation of early ovarian follicles，This experiment was conducted to study the Tissue Expression Pattern，mRNA expression levels and SNPS. We use RT-PCR,real time PCR and PCR-RFLP analyse the Tissue Expression Pattern,the mRNA expression levels in the ovary of single fetuses and triplets Hu sheep during oestrum,and the polymorphism at 152 on Exon1 of GDF9 gene.The results indicated that GDF9 gene expresses in Hypothalamus, pituitary, ovary,uterus, Fallopian tube, heart, liver, spleen,lung and kidney of Hu sheep；the mRNA expression levels on GDF9 gene in the ovary of triplets Hu sheep is prominent higher than that in single fetuses Hu sheep(P < 0.05),the single nucleotide mutation:A→G at cDNA 152 of GDF9 gene in Hu sheep is not detected.The analyse results indicated that The mRNA Expression Level of GDF9 gene have some relation with the reproduction power in Hu sheep,and wait for the further analyses of The polymorphism of GDF9 gene in Hu sheep.

Purpose: To obtain mimicking epitopes of mAbs against infectious bursal disease virus (IBDV) VP2 using phage display technique, so as to offer basis for further study of their immuno-protection against infectious bursal disease (IBD). Methods: The 12-mer phage random peptide library was screened with four purified mAbs against IBDV-VP2（1B5、5D1、2H11 and I-4-4-3 ) and 40 clones were obtained, among which, 32 positive clones were identified by ELISA.We chosen 20 positive clones for DNA sequence analysis and four ascendant 12-peptides were determined as different epitopes of IBDV-VP2. The four ascendant 12-peptides shared no more than three continuous amino acid residues similar with the sequences of IBDV-VP2 registered in GenBank, and their conjunction with mAbs against IBDV-VP2 can be inhibited by VP2 protein,which indicated that the epitopes were conformation-dependent. When the four mimicking epitopes were expressed tandem, protein r4EPIS was obtained. The results from SDS-PAGE analysis showed that the proportion of r4EPIS was 20% of the total bacterial proteins and the molecular weight was 30 kDa. The r4EPIS showed specificity and reactionogenicity by use of immunoblotting test with polyclonal antibodies. Conclusion: Four conformation-dependent mimic epitopes of mAbs against IBDV-VP2 were obtained by phage display technique.

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Abtstact：The elimination of marker genes after selection is a main premise for the commercial use of genetically modified plants, and site-specific recombination can be used to eliminate maker gene. There are three main site-specific recombination systems which can be used successfully in plants: Cre/loxP, FLP/FRT and R/Rs. To remove selectable marker genes from transgenic plants by the site-specific recombination system, there are generally several strategies: constitutive expression of recombinase by re-transformation of Cre gene or crossing with Cre transgenic plant, or application of transient, inducible or tissue-specific expression of recombinase, among which inducible autoexcision is the most potential way in plant transformation.

Currently, the research on cell wall is one of hot researches in plant molecular biology. Xyloglucan (XG) is the main hemicellulose of primary cell wall in dicotyledon. Xyloglucan endotransglucosylase/hydrolase (XTH) is a family of enzymes that catalyze xyloglucan endotransglucosylase (XET) and/or xyloglucan endohydrolase. XTHs can cut XG chains and transfer the fragment with the new reducing end either to another XG or to water, and play an important role in cell wall remodeling, affecting plant growth and development. In this article, we reviewed the structures and functions of XTHs, and the factors that affect their functions. The current problems and future research directions were outlined.

This experiment was conducted to identify two nontargeted bands accompanied with heterozygote in nondenatured polyacrylamide gel electrophoresis. The nontargeted and targeted bands were cut and reclaimed, and used into PCR as templates. The PCR products were electrophoresised according to different combinations and disposals. In addition, 4 synthesized single strands were applied to simulate the formation process of nontargeted and targeted bands. The results showed that 1) The mobility property of the PCR products of DNA extracted from untargeted bands was the same as that of the PCR products of genomic DNA from heterozygote. The mobility property of the PCR products of DNA extracted from either targeted band was the same as itself. 2) If the PCR products of DNA extracted from the two targeted bands were mixed and treated with denaturation and renaturation in PCR instrument, the mobility property of them was also the same as that of the PCR products of genomic DNA from heterozygote; if mixed without other treatment, there were only two targeted bands appeared. 3) In the simulation experiments, the untargeted bands were formed by the heteroduplex DNA strands and the targeted bands by the homoduplex DNA strands. These results indicated that the two nontargeted bands were not nonspecific amplicons. Because of a 3bp deletion in one of two targeted double-stranded DNAs, two heteroduplex DNAs formed in the processes of denaturation and renaturation of PCR for random pairing of single-stranded DNAs, thus, convex rings were produced consequently. The convex rings decreased the mobility of the two heteroduplex DNAs and the two delayed heteroduplex DNAs formed the two nontargeted bands.

BALB/c mice(Mus musculus) were immunized five times with purified recombinant rGST-BoIFN-γ protein expressed in E.coli BL21(DE3) at 100μg per mice. Murine myeloma cells were fused with splenocytes of the immunized mice. An indirect ELISA with recombinant rHIS-BoIFN-γas antigen was used to screen antibody-producing hybridomas. Two hybridomas cells could produce McAbs steadily after 3 cycles of subcloning, all McAbs showed positive reaction to rHIS-BoIFN-γ in western blot. All McAbs were IgG1 isotype and κ chain. The two McAbs can be partner for detect antigen.