Reproductive traits are important economic traits in sheep. This review introduced bone morphogenetic protein receptor IB, bone morphogenetic protein 15 and growth differentiation factor 9 genes and their relationships with high prolificacy in sheep, and application of both FecB and FecXI genes.

Abstract BALB/c mice were immunized intraperitoneally with purified recombinant mEIFN-protein. Murine myeloma cells were fused with the splenocytes of the immunized mice after the fourth immunization. An indirect ELISA with recombinant mEIFN-derived from the baculovirus expression system as antigen was used to screen antibody-producing hybridomas and 15 McAbs against mEIFN-were obtained. Twelve hybridomas cells of them can produce McAbs steadily after 3 cycles of subcloning. All the 12 McAbs were reactive in (indirect fluorescence assay) IFA tests, and all of them showed positive reaction to mEIFN-. Four McAbs are IgG1, two McAbs are IgG2a，three McAbs are IgG2b and three McAbs are IgM isotype, respectively. The light chains of all McAbs are chain. The seven McAbs of them showed positive reaction to GST-mEIFN-(1-84) protein, and the others show positive reaction to GST-mEIFN-(80-146) protein in specific ELISA tests, respectively. The McAbs against mEIFN- protein developed in this study would be useful as a basis of detection methods for equine IFN-, investigating the structure and biological effects of equine IFN-.

The full length of 3AB gene of foot-and-mouth disease virus (FMDV) was amplified and subcloned into pET-30a vector by two unique restriction sites. The recombinant plasmid pET-3AB was transformed into a BL21 (DE3) strain of E.coli. The recombinants were induced with IPTG at 37℃and 28℃ for expression of target protein. The product was analyzed by SDS-PAGE. The results showed that the expressed protein was solubly expressed when induced at 28℃ and formed inclusion body when induced at 37℃. Both products could reacted well with sera derived from FMDV infected cattle by western blotting. The expressed product was further purified and used as the antigen in an indirect ELISA assay for detection of different animal sera. The result showed that the partially purified 3AB protein could only react with sera derived from FMDV infected animals. This preliminary result indicated that the expressed 3AB could be used as antigen in ELISA for discrimination FMDV naturally infected animals from vaccinated animals.

The general combining ability (GCA) and specific combining ability (SCA) of 6 selected optimum parents of Betula platyphylla Suk in a P(P-1)/2 diallel cross were estimated by analyzing two growth phenotypes of their offspring. The results showed that two phenotype were controlled by additive effect and non-additive effect simultaneously. The general combining ability of the different phenotypes of the same parent has significant difference, and the general combining ability of the same phenotype between different parents also has significant difference. Parent Q2 is the best one, M2 take second place. Moreover, the specific combining ability of the same phenotype in various cross combination and the two phenotype of the same combination also vary prominently. After investigation and statistical analysis, we found the cross 8×Q1 has the highest SCA of height and ground diameter, and the GCA of height and ground diameter were higher than SCA. The heritability of height and ground diameter were strongly heritable, suggest that they can be used for early selection.

Site-directed mutagenesis to replace residue 32 of Musca domestica Defensin gene (glycin, G) with arginine (cationic amino acids) by recombinant PCR, In this way, the first mutant des-hf-MU1 was gained. Six lysines (K) were added to the C terminal of des-hf gene in the same way, forming the second mutant des-hf-MU2.The two mutant were expressied in Pichia expression system, The expression product showed perfect autibacteril activity against gram-postive bacterias.According to Agarose Diffusion Assay, the anti-bacteria activity of des-hf-MU1 is 2.4 times than des-hf, and des-hf-MU2 is 8.0 times than that. Moreover, the activity is hightest when PH is 5~6, heat stability experiment showed that the antibacterial activity still exist when des-hf-MU1 and des-hf-MU2 were heated to100℃.Above all reveals sufficiently that ds-hf-MU1 and des-hf-MU2 have good perspective in application.

Using in vitro hypocotyls as explants, the adventitious shoots of Euonymus fortunei var. Radicans were differentiated directly from the basal MS medis supplemented with different plant growth regulators through organogenesis. The results showed that the highest regeneration frequency was obtained with MS medium containing 0.5 mgL-1 6-BA and 0.01 mgL-1 NAA. Ninety-two percent of regeneration frequency and 4.2 shoots per explants were obtained after 30 days of culture. The binary vector pBCGm containing Galanthus nivalis agglutinin (GNA) gene was introduced into Agrobacterium tumefaciens LBA4404. The hypocotyl segments of Euonymus fortunei var. Radicans were infected through Agrobacterium tumefaciens. The GNA gene was integrated into the genome of transgenic plants confirmed by PCR and PCR-Southern analysis. The highest transformation frenquency was obtained through the following transformation procedure after preculture for 0 d, the explants were infecter for 30 min with OD600=0.6 Agrobacterium tumefacien, and the co-cultivated for 3d.

To identify Hemagglutinating Encephalomyelitis Virus (HEV) 67N Receptor in PK Cells Membrance, express S1 protein of HEV in Pichia Pastoris, and purify the protein by Ni2+ affinity chromatograph. Polyclonal antibody to HEV was prepared with immunizing rabbits by injecting the purified S1 protein four times. After SDS-Polyacrylamid gel electrophoresis (SDS-PAGE), the PK cells membrance proteins were transferred to nitrocellulose membrance. VOPBA was performed by the recombinant S1 protein to identify the receptor protein binding HEV-S1. The result showed that HEV-S1protein bound one band about 90 kDa in PK cells membrance. This result is very important rationale to study thoroughly the pathogenic mechanism of Hemagglutinating Encephalomyelitis Virus.

Mature microRNAs（miRNAs）are a class of non-coding RNAs with ～21 nucleotides in length. The majority of miRNAs matches perfectly to 3′UTR sequences of mRNAs and result in mRNA degradation. They also regulate target genes by binding imperfectly to sequences in mRNAs and attenuate translation. Functional analysis indicated that miRNAs involved in a wide range of pivotally biological events, such as growth and development, regulation of immune response, virus infection, etc. In this study, we compared two approaches in miRNA identification. MiRNAs let-7b and let-7c was cloned and sequenced in porcine skeletal muscle tissues.

To clone chicken RANKL(chRANKL) gene by RT-PCR method and obtain the chRANKL protein expressed in E.coli and analyse its bioactivity. The encoding sequence of chicken RANKL was amplified from embryo chicken osteoblasts total RNA by RT-PCR. The PCR product was inserted into the prokaryotical expression vector pET-32a(+). The recombinant vector was transformed into E.coli BL21(DE3)Plys and IPTG inducement. Then the purified protein was added to coculture with the mature chicken osteoclasts in vitro to observe its activation. Result showed that the PCR product was about 1200 bp long, which was consistent with the expected one. The sequence of insert corresponded with the published encoding sequence of chicken RANKL. Plasmid pET32a(+)-chRANKL was transformed into E.coli BL21(DE3)Plys and a 64 Kd protein was expressed by IPTG inducement, and Western blotting indicated that recombinant protein had good antigenicity. We also found that the chRANKL could stimulate the activity of mature osteoclast which increased both the number and the area of bone resorption lacunae relative to dose-dependent manner.

Recombinant expression vector k14/c-myc-paav was constructed bearing the mouse c-Myc cDNA derived by the skin specific human keratin 14 promoter, then the linear vector was microinjected into the two pronucleus of the C57BL/6 fertilized eggs to produce transgenic mice。256 microinjected eggs were transferred, 6 of the foster mothers were pregnant and 44 adult mice were produced, two of them were positive transgenic mice confirmed by PCR and Southern blot analysis, transgenic rate was about 4%. Southern blot results showed that the foreign gene integrated in mouse chromosomes with multi- inverted repeat sequences.

Porcine GM-CSF gene was amplified from porcine expression plasmid pGM-CSF by PCR. Fragment containing GM-CSF gene was subcloned into somatostatin eukaryotic expression plasmid pcS/2SS to construct fusion-expression plasmid (pGM-CSF/SS) harboring porcine GM-CSF and somatostatin (SS). Restriction endonucleases digestion and DNA sequencing showed that the fusion-expression plasmid (pGM-CSF/SS) harboring porcine GM-CSF and somatostatin was successfully constructed. To study the immunization effect of pGM-CSF/SS on mice, a total of 30 mice at the age of 22 days were divided into 3 groups and immunized with salt solution (group 1), pcS/2SS (group 2) and pGM-CSF/SS (group 3), respectively. The results showed that lymphocyte proliferation response of pGM-CSF/SS was higher than that of the control group and pcS/2SS alone (P<0.05), indicating that GM-CSF could enhance lymphocyte proliferation responses. Fusion-expression plasmid of GM-CSF with SS significantly enhanced the P/N value of SS antibody compared to the control group and immunization of pcS/2SS alone. Percentage of mice with positive SS antibody immunized with pGM-CSF/SS was higher than that of other groups, which got to 60%. Percentage of that immunized with pcS/2SS was 30%. In conclusions, genetic adjuvant GM-CSF can enhance immune response of somatostatin DNA vaccine, which is an access to further research for immuno-enhancement of GM-CSF gene on somatostatin DNA vaccine.

Three pairs of primers were designed to detect single nucleotide polymorphisms of partial 3′untranslated region (UTR) of bone morphogenetic protein receptor IB (BMPR-IB) gene in four sheep breeds (Small Tail Han, Hu, Texel and Chinese Merino sheep) by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP). Only the products amplified by primer P1 displayed polymorphisms. For primer P1, two genotypes (AA and BB) were detected in Hu sheep, and only one genotype AA in the other three breeds. Sequencing revealed two mutations（121T→C and 195T→C）of BMPR-IB gene in the genotype BB in comparison to the genotype AA.

In this study, the part of halothane gene in 18149-18760 sites in 94 zang pigs was amplified by PCR. The sequence of PCR production is determined. The result shows that PCR production is 612 bp, and it included exon 17 and cDNA 1843 site. The result of HhaⅠpolymorphism analysis about this 612bp was all 94 zang pigs showed halothane masculine. Sequence analysis results indicated that two mutation sites are found in the sequences of the four zang pigs, and all mutation sites are transition and locate within introns in all 4 sequences. In intron 16 one site has transition heterozygote. The C1843→T1843 mutation site is not found in zang pigs.

Abstract: The Taqman Fluorescent probes and primers were designed and synthesized according to the conserved part of hypothetical functional protein gene of Mycoplasma suis (Eperythrozoon suis) available in GenBank，and then reaction parameters were optimized and standard carve was established based on the quantitative 10×series of dilutions recombinant M.suis plasmid （pGEX-T/M.suis）to develop a real-time TaqMan-fluorescent-quantitative PCR assay (FQ-PCR) of Mycoplasma suis. The sensitivity, specificity and repetition assay of FQ-PCR were tested, and 24 heparinized-blood samples taken from suspicious infected pigs basing on the clinical signs and microscope check were detected by using the established FQ-PCR assay comparing with the routine PCR method. The results indicated the cycle numbers of standard carve had a good linear relation with the template concentration, and the correlation coefficient was 0.998. The developed FQ-PCR assay could detect 1.0×101 copy/μL of plasmid DNA and its sensitivity was 100 times higher than that of the routine PCR. The sensitivity assay exhibited that the positive carves could be achieved from the pGEX-T/M.suis recombinant plasmid, whereas the negative carve were displayed from the genomic DNA of the other 8 kinds of pathogenic microorganism acting as the control. Three pGEX-T/M.suis recombinant plasmids samples with different concentrations were examined using the FQ-PCR repeated 2 times and the results indicated that the FQ-PCR was reproducible. The FQ-PCR was used to detect the DNA of 24 suspected heparinized-blood samples and 20 positive samples were displayed, which showed the better sensitivity than that of the routine PCR , with 19 positive samples from the same 24 suspected samples.

The purpose of this study was to evaluate the effects of donors’ sex, cell cycle synchromization treatments and donor nuclei obtained from fresh or frozen-thawed on in vitro developmental competence of yak-bovine interspecies nuclear transfer(NT) embryos. In this paper, we used bovine oocytes as recipient, yak ear fibroblast cell as donor. The results indicated that the development rate of male blastocysts was higher than female（56.6% vs 39.5％，P﹤0.05）, whereas cleavage and total cell number had no difference between the two groups. No significant difference was observed in the blastocysts development and quality of donor treat with serum starvation or confluence inhibition. There was no significant difference in embryos development of use fresh or frozen-thaw donor cell, whereas cleavage in group of fresh donor cell was significantly higher than frozen-thaw group（54.5％ vs 78.2％，P﹤0.05）. Our results demonstrated that donors’ sex could impact the in vitro developmental competence of yak-bovine interspecies NT embryos, whereas different cell cycle synchromization treatment and freezing had little influence.

In this study, the in-vitro maturation (IVM) and in-vitro fertilization (IVF) of ovine oocytes with inhibitor, BL-I, was investigated. Oocytes were cultured in vitro (IVC)among different durations after being inhibited for 8h. Results: MⅡ rate of the 8h IVC group was significantly lower than the other groups. There were no significant differences of the maturation rates among the 16h、20h IVC groups and the control one; the maturation rate (70.83%) of the 24h IVC group was higher than the control one (66.18％) ,but there were no significant differences between them (P>0.05). There were significant differences of the blastocyst rates among the 16h、20h IVC and control groups . Conclusion: The effect of inhibition by BL-I on IVM of ovine oocytes is reversible; and the inhibition treatment with BL-I on ovine IVM, can not improve the fertilization in-vitro and developmental competence of embryos.

This experiment focued on investigating affects of different supplements in the media containing KSR, FBS, FBS+PD98059, respectively on embryo attachment, formation of ICM outgrowths, isolation and culture of embryonic stem cells (ESCs) from mouse embryos of Kunming species, which was used as the experimental material while mitomycin C-inactivated mouse embryonic fibroblasts (MEF) as the feeder layers. The results demonstrated that the efficiencies of embryo attachment in the medium supplied with KSR was significantly lower compared with that of the medium supplied with FBS（P＜0.05), and formation efficiencies of ICM outgrowths and ESCs clonies at the first passage appeared no significant difference between them (P>0.05), however, formation efficiencies of ESCs clonies at the 2~5 passages were significantly higher than those in the medium supplemented with FBS (P＜0.05), two ESCs lines were subcultured to the seventh passages. Meanwhile, the efficiencies of embryo attachment, formation of ICM outgrowths and ESCs clonies at the 1-5 passages in the medium supplemented with FBS + PD98058, were significantly lower than those in the medium supplemented with KSR and FBS, respectively（P＜0.05). Providing ICM cells and ESCs were trypsinized with 0.5g/L trypsin +0.2g/L EDTA combined with manual dissection, ESCs clonies at the 1~5 passages appeared significantly higher than those with 2.5g/L trypsin + 0.2g/L EDTA （P＜0.05). Therefore, the results indicated that it was advantageous to isolation and culture of ESCs from mouse embryos of Kunming Species in the medium supplemented with KSR over FBS and FBS+PD98059, respectively. Accordingly, the ICM cells and ESCs at the initial passages were trypsinized more properly with 0.5 g/L trypsin associated with manual dissociation, than with 2.5 g/L trypsin.

The reproductive function of three strains of mice (Kunming, ICR and C57BL/6-Tg(CAG-EGFP)C14-Y01-FM131Osb) oocytes reconstructed by the nuclei of first polar body (abbreviated to PbI) was explored in these experiments, respectively. Cumulus oocyte complexes (COCs) from the mouses were collected after superovulation, then PbIs were obtained from the COCs by 2% pronase treatment. The survival PbIs under different temperature was identified and stapled by the methods of morphology and trypanblau staining, later the contents including nuclei and a little cytoplasm of PbI was injected into the oocyte that its nuclei had been enucleated by micromanipulation before, the reconstructed oocytes by PbI was fertilized and cultured in vitro in order to observe its ability of further development. The results showed that vigor of PbIs was in the best status within 12 to 14 hours after superovulation, and still maintained during 48 hours under 4℃. 13 out of 117 reconstructed oocytes from Kunming and ICR mice developed into 2-cell embryos, and 3 out of 38 reconstructed oocytes from C57BL/6-Tg(CAG-EGFP)C14-Y01-FM131Osb mice was cleavage as 2-cell embryos. The experiments confirmed that the mouse oocytes reconstructed by the nuclei of PbI could be normally maintained its fertilization and further development, this has offered valuable reference for further utilization of gene resources for other mammalian animals.

Abstract: Rat hepatocytes were treated with clofibrate, the express levels of PXR and CYP3A1 were determined by Western blot or RT-PCR. The results as follows: Clofibrate markedly increased the expression of PXR, and the induction was abolished by actinomycin D, an inhibitor for mRNA synthesis. No induction of CYP3A1 by clofibrate was tested, PCN alone markedly induced the expression of CYP3A1, co-treatment with clofibrate enhanced the induction. Clofibrate was one of pero瑇isome proliferators, the induction of PXR and CYP3A1 by clofibrate provided a molecular mechanism on drug metabolism.

Medicine produced by transgenic microorganisms and plants would have the potential to change the traditional means of production, and to reduce the cost of medicine production. In this paper, a growth hormonegene (AmGH) from giant panda (Ailuropoda melanoleuca) was cloned, and pCAMBIA13011-GH, the plant expressing vector consisting of Giant Panda (Ailuropoda melanoleuca) growth hormone gene was constructed, which contains a CaMV35S promoter and a hygromycin resistance gene, and then transformed AmGH into tobacco leaf-disc by Agrobacterium-mediated method. A total of 50 regenerated plants with hygromycin resistance were obtained in the selection media. The 12 putative transgenic individuals were tested and 5of them were verified the presence and integration of the AmGH into the nuclear genome of tobacco plants by GUS, PCR analyses. Expression of AmGH gene has been confirmed in E coli and in plants. To our knowledge, this is the first report of the expression of Ailuropoda melanoleuca growth hormone gene (AmGH) in E coli and plant system, suggesting a major step in the production of plant-based for Ailuropoda melanoleuca growth hormone (AmGH)

We investigated a transformation system to breed a new pollenless material of Lilium Longiflorum X L. formosanum. The results indicate that using in vitro bulb scales as explants, regeneration plants were obtained with 100% of regeneration rate through organogenesis on the basal MS media supplemented with 1.0 mg/L 6-BA and 1.0 mg/L NAA. A system for the production of transgenic plants was developed for the Lilium Longiflorum X L. formosanum by Agrobacterium-mediated transformation. The highest transformation efficiency was obtained through the following transformation procedure, after pre-culture for 3 d, the explants are immersed into bacterial suspension (OD600≈0.8) for 5 min, and then co-cultivate for 5 d. Using this system, a maize pollen specific gene, zm401, was integrated into the genome of the lily and was confirmed by PCR and PCR-Southern analysis. The results can be as a previous work to obtain indirect materials of pollenless lilies.

Production of transgenic sweetpotato plants expressing the bar gene for herbicide resistance was achieved using Agrobacterium tumefaciens-mediated transformation and sweetpotato cv. Lizixiang embryogenic suspension cultures. Total 450 embryogenic cell aggregates were cocultivated with the A. tumefaciens strain LBA4404 harboring a binary vector pCAMBIA3300 with the bar gene. Eight weeks after selection on MS medium supplemented with2.0 mg/L 2,4-D, 100 mg/L Carb and 0.5 mg/L PPT, 19 PPT-resistant embryogenic calluses were produced. After transferred to MS medium supplemented with 1.0 mg/L ABA, 100 mg/L Carb and 0.5 mg/L PPT, 10 of them formed 103 putatively transgenic plants. PCR analysis indicated that 69 plants were transgenic. Stable integration of the bar gene into the genome of transgenic plants was confirmed by Southern blot analysis. Transgenic plants exhibited functional expression of the bar gene by in vivo assay for herbicide resistance.

In plants, acid invertases are known to be the key enzymes cleaving sucrose into reducing sugars (RS) (glucose and fructose). To improve quality of potato (Solanum tuberosum L.) fries which is largely influenced by accumulation of RS in tubers stored at low temperature, a part of acid invertase cDNA with hpRNA structure was transformed into potato cv N2. Detections of PCR amplification and Northern Hybridization suggested that the RNAi vector was successfully transformed into cv N2. Analysis of acid invertase activity of the plantlets and microtubers of RNAi transgenic lines indicated that expression of the interference RNA repressed significantly the activity of acid invertase of plantlets by an average 69.8% (excepted with Line Ni-1) and led to the best decrease of 78% (Line Ni-4), and the highest decrease of that of microcoubers by 68%. Compared with that of well inhibited antisense inv transgenic plants, the comparative downregulation of RNAi suggested a distinct alteration on endogenous acid invertase and a potential strategy of PTGS in modulation of cold-sweetening in potato.

Seven hundred and ninety one microsatellites(SSRs) were isolated from 7055 Panax ginseng expressed sequence tags(ESTs). According to primer design criteria, sixty-eight primer pairs for EST-SSR were designed. Under an appropriate PCR reaction system, all EST-SSR primer pairs were screened against genomic DNA of ji’anchangbo and fusong’ermaya from panax ginseng respectively, and forty-three of sixty-eight EST-SSR primer pairs resulted in PCR products. Then all forty-three EST-SSR primer pairs available were tested on nine panax ginseng varieties, two panax quinquefolius varieties and two acanthopanax senticosus varieties for polymorphisms, and twenty-six EST-SSR primer pairs were polymorphic, accounting for 60.47% of primer pairs amplified, accounting for 38.23% of total primer pairs designed. These results showed that it is an effective and feasible approach to develop EST-SSR markers in accordance with ESTs in panax ginseng.

The open read frame of ap36 gene was amplified from plasmid pMYE by PCR using a pair of special primers. The PCR production was cut by XbaI and SphI and sub-cloned into the pBMB982-304. Thus, the ap36 gene sequence located the downstream of slh motif of S-layer protein gene. Then the recombinant plasmid pBMB0588 was transformed into Bacillus thuringiensis plasmid–free derivative strain BMB171. The result showed the target protein was successful expressed as a SLH-Ap36 fusion protein. In order to investigate the induced ability of the recombinant strain, tomato and potato chips treated with the 24h cultured of recombinant strain were investigated to determine the induction of defense responses. The result showed the activity of peroxidase and the amount of proline of tomato leaves was increased 57.14% and 131.59% compared to control after treatment of 90 min, respectively. The activity of phenylalanine ammonia lyase was also higher than control after treatment of 4d. Furthermore, the treated potato showed high resistance to rot disease caused by Erwinia corotovora SCG1 compared to control.

In a 7 L fermentor, the production conditions of engineering strain NB01 containing chitinase gene were investigated. By enhancing aerati0 and changing the agitation rate，the DO level was higher than 30%. The methanol feed rate was varied with the change of DO level in order to avoid the methanol concentration too higher to injure the cells. And the results showed that the maximum chitinase activity could reach 48.0795 U/mL at 84h. The fast purification of recombinant chitinase was conducted with Hollow fiber UF membrane and Sephadex G-100 and the SDS-PAGE analysis indicated that there is a single band with the right molecular weight corresponding to the chitinase on the gel. Furthermore, the recombinant enzyme is stable at a wide range of temperature and pH. Also, it is rather stable when stored at 4℃ for a long time.

Based on sequence analogy and amphipathicity analyse of α-helical antimicrobial peptides, a sequence template was extracted. The model Antimicrobial Peptide PGYa(peptide beginning with Gly and ending with Tyr-NH2) was designed by virtue of the template and computer-aided design subsequently. Using the optimal condon of Pichia pastoris, PGYa gene with 94bp length were achieved through a recursive PCR strategy. Especially a Kex2 signal cleavage site was fused in 5’end of the antibacterial peptide gene to make sure the expression protein with nature N-terminal. The modified antibacterial peptide gene was then cloned into the pPICZα-A vector to construct the recombinant expression vector pPICZα-A- PGYa, and transformed into yeast host strain X-33. Under the control of the promoter AOX1(alcohol oxidase 1), the PGYa protein with a molecular weight of 2.8 KD was expressed in supernatant and the amount of secreted pertein can reach 132 μg/mL. Preliminary antibacterial assays showed that PGYa has a potent antibacterial activity against E.coli DH5α.

The preparation condition of probiotics-loaded montmorillonite integrated the probiotics and crude silicate montmorillonite was optimized and its influencing factors research was also discussed. The results showed that number of probiotics attached to montmorillonite was invariablenes at montmorillonite weight of 0.5 g and increased with the initial concentration and volume of probiotics. The adsorption ability decreased with the increasing pH values and presented the same trend in different ionic concentration. At different temperature, the adsorption rate of montmorillonite to probiotics increased firstly and then decreased and the highest adsorption percent was up to 89.74% at 30℃. This research indicated that the strains adsorption depended on not only the biochemical and surface characters, but also the environmental condition such as temperature.

Type II secretory system is associated with secretion of pathogenic proteins of Ralstonia solanacearum. The gsp gene cluster encoding type II secretory system of R. solanacearum consists of 12 gsp genes. All of gsp genes were cloned successfully by designing optimal PCR reaction system suitable for high content of GC. The 12 gsp genes were ligated into bait and prey vector, respectively. The bait and prey pool of yeast two-hybrid system were constructed. The open reading-frames of 12 gsp genes were verified by sequencing to ensure that gsp genes can be expressed rightly in yeast system, which establish the foundation for research interactions between GSP proteins.

Abstract: In the present paper, the genetic diversity 6 kind of 48 cotton trains from different origins were estimated using inter simple sequence repeat (ISSR) markers. 11 among 60 ISSR primers were chosen for their reproducibility and high polymorphism. a total of 90 fragments were amplified, in which the percentage of polymorphic loci (PPL) was76.1%, Results of POPGENE analysis indicated that genetic diversity of Xinjiang population was the highest (PPB= 91.43%, H= 0.3769, I= 0.5464),and Zhongmiansuo, Henan population followed it and genetic diversity was near among other three population. Analysis of Nei’s genetic diversity index indicated that the intraspecific genetic diversity were 82.42% ,diversity among populations were 19.72%; the gene differentiation (Gst) was 0.1759 and the gene flow (Nm) was 2.34 ;Result of UPGMA cluster analysis indicated that the relationship between Shandong and Shanxi is nearest It were the first clustered with Hebei, and Henan and CCAS were followed successively, Xinjiang population were clustered finally.

The phytohormone abscisic acid (ABA) is not only involved in the plant abiotic stress response but also plays a major role in the regulation of various biotic stress responses. ABA negatively regulates plant biotic resistance which is through the interaction with the signal transductions of slicylic acid, jasmonic acid and ethylene, and shared components in pathogen-related signaling. In this paper, we review the signal transduction of ABA and its regulatory role in biotic responses.

Efficiency and specificity, are the limiting factors in preparation of transgenic animal. This review describes the recently developed animal gene transfer techniques, including gene transfer into the testis, ovary and RNA interference for improving efficiency of non-site specific methods; gene targeting in embryonic stem cells, somatic cells and primordial germ cells for site specific methods; and site- and time-specific gene targeting and controlled expression of transferred genes. The merits and disadvantages for each method are also discussed, as well as potentials of using these methods to develop transgenic animals.

Partial genome of Tobacco vein distorting virus（TVDV）was amplified from total RNA extracted from tobacco plants infected with tobacco bushy top disease by RT-PCR. The amplified fragment comprises 1655bp，which contains partial RNA-dependent RNA polymerase（RdRp）gene，complete coat protein（CP）gene and complete movement protein（MP）gene. The intergenic region of RdRp gene and CP gene consists 201 nucleotides，which are identical in arrangement to the genus Polerovirus. Molecular homology trees were constructed based on the amino acid sequences of the RdRp，CP and MP，and the results showed that TVDV shares much more homologous with the specieses of the genus Polerovirus than other genuses of family Luteoviridae，which indicated that TVDV should be a definitive member of the genus Polerovirus in the family Luteoviridae.

A basic heme-peroxidase (WsP) was purified to homogeneity from wheat (Triticum aestivum) seed by consecutive treatments consisting of ammonium sulfate fractionation，ion exchange chromatograph On FPLC-Source30s and gel filtration chromatograph on FPLC-Superdex G75。The specific activity of purified peroxidase was 3912 U／mg，The protein was exhibited a molecular mass of 37kDa.．Spectral analysis of the enzyme revealed the presence of the Soret band with a max at 399 nm。 The most adaptive pH and optimum temperature of peroxidase was pH5.0 at and 40℃。 The best oxidation concentration of H2O2 for peroxidase was 14 mM/L。