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本期目录
2008 Vol. 16, No. 4 Published: 24 July 2008
研究论文
Expression and Bioactivity Characterization of NP-1 in Transgenic Nitrate Reductased Deficient Mutant of Chlorella ellipsoidea
2008, 16(4): 0- |
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Abstract:It is a potential valuable way using transgenic Cholrella as a bioreactor to produce alien proteins. In this paper, the vector containing the NP-1 gene under the control of the Ubiquitin promoter and two selective marker genes NPTII and nitrate reductase gene was constructed. The NP-1 gene was transformed into the nitrate reductase-deficient mutant nrm-4 of C. ellipsoidea via a protoplast transformation method. The transformed nrm-4 cells were screened on the medium with nitrate and G418. The transgenic lines were characterized by PCR amplification of both the NP-1 gene and NPTII genes from genomic DNA isolated from transformants. The transgenic nrm-4 was further cultured in the liquid medium with nitrate but without G418. In vitro assay results showed that the protein of the transgenic C. ellipsoidea had effective bacteria- resistance activities and it is stable without G418 selection stress. The research provided a new way to express alien genes in Chlorella, and the basis to largely produce NP-1.
Expression and Purification of HPT gene in Escherichia coli and Preparation of Its Polyclonal Antibodies
2008, 16(4): 0- |
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Hpt gene was cloned from genetically modified plants’ genome and digested by BamHⅠ和Hind III. pET30a vector was digested by the same enzymes and ligated to hpt gene to construct plasmid pET30a-hpt. HPT protein was expressed by E.coli BL21 and purified firstly by ammonium sulfate, then by Ni+-NTA column. Hygromycin B phosphotransferase (HPT) was highly expressed in Escherichia coli BL21(DE3) in the presence of isopropyl-β-D-thiogalactopyranoside (IPTG) and most products existed in soluble form. After cleavage of His•Tag fusion protein by enterokinase, intact、biologically active HPT was released from the fusion protein and was purified to homogeneity with Ni2+ affinity chromatography. Polyclonal antibodies against were raised in three rabbits and the purified polyclonal anti-RCT antiserum appears a very high degree of sensitivity and specificity.
Detection and pathogenic effect of quorum sensing signal molecule of Acidovorax avenae subsp. citrulli
Tao Chen Guoliang Qian Xiaoli Yang Junyi Ma Fengquan Liu
2008, 16(4): 0- |
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Bacterial Fruit Blotch (BFB) of melons is one of the most serious diseases of both native and overseas water melon and muskmelon, while up to now, there is still no effective control method except strict quarantine measures. With the discovery and study progress of quorum sensing (QS), using signal interference technology to control plant bacteria diseases has been a research focus and also successfully applied. This article firstly detect signal molecules (AHL) from melons Bacterial Fruit Blotch pathogen (Acidovorax avenae subsp. citrulli) by bioassays method. Gene aiiA which can degrade AHL molecules was transformed into BFB pathogen strain NJF10. Inoculation tests showed that this transformed strain has a dramatically reduce of pathogenicity.This result lays a foundation for controlling melons Bacterial Fruit Blotch by signal interference strategy.
Increasement of Maize Amylose Content through RNA Interference
Xin-mei GUO
2008, 16(4): 0- |
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To regulate the biosynthesis of maize amylose, an expression vector of RNAi with the fragment of sense gene +FAD2 intron +antisense gene had been constructed, and. was transformed to maize calli which were induced to form from the elite inbred lines by Biolistic PDS1000/He. The selection was carried out on the media containing concentrations of xylose. 12 transformed plantlets were obtained. The integration of alien gene was conformed in 6 transformed plantlets with the analysis of DNA dot blotting, PCR and PCR-Southern hybridization. The seeds amylose content of regeneration plants was detected,and amylose content of a transgenic line increased up to 15.6%.
2008, 16(4): 0- |
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Abstract: A Nicotiana tabacum cDNA library consisting of 5927 high quality sequences was sequenced. By analyzing the EST sequences, the proportion of Nicotiana tabacum genes were identified at EST level. A cDNA array was constructed based on the TUTs derived from EST assembling results. A total of 42 differentially expressed genes were identified including Plasma membrane intrinsic protein 2a, Ethylene-responsive proteinase, and Pre-mRNA splicing factor prp31 gene etc., suggesting a complicated biological processing in Nicotiana tabacum under Saline Stress.
Development of PCR-ELISA for identification of Listeria monocytogenes
Bing-Qian Tan Qi-gai He Huan-chun Chen
2008, 16(4): 0- |
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Objective to detect Listeria monocytogenes in foods, a simple PCR method and PCR-ELISA method were developed, according to hlyA gene sequence clone and analysis, specific PCR primers labled with biotin in 5?end of first primer and a specific probe labled with digxin in 3?end were designed by primer 5.0. The results showed that only L. monocytogenes strains were successfully amplified the expected fragment, whereas the E.coli, Salmonella, L.iuanuii, L.innocua, L.innocua, L.seeligeri, and L.gray were negative. Simple PCR method followed by agarose gel electrophoresis and staining of PCR products with ethidium bromide, while PCR-ELISA detected amplified products wih an enzyme-linked oligosorbent assay determinat optical density value at 405 nm. 60 foodstuff samples were further screened to validate the PCR-ELISA procedures by the results compare to the isolation method SN/T0148.1-2005. Positive proportions of molecular methods revealed higher sensitivity than isolation and identification method. It抯 showed that PCR-ELISA improved sensitivity and specificity, and the data revealed the sensitivity of detemination of PCR products 150 folds than agarose gel electrophoresis.
The difference analysis of two α-amylase from Bacillus cereus and gene clone expression
Qi WANG
2008, 16(4): 0- |
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Bacillus cereus can produce α-amylase which has important industrial value and can endure high temperature. The enzymatic characteristic of two α-amylase from Bacillus cereus (905 and 904)were studied. The results showed that: the activity stability temperature of both α-amylase is 90℃-100℃. Their thermal stability and enzymatic activity do not depend on Ca2+. But their protein molecular weights are obviously different. The structural gene of amy905 and amy904 were successfully cloned by PCR and were successfully expressed in E. coli. The experiments showed that, the gene of amy904 is 1362bp and the protein molecular weight is about 55 KD; the gene of amy905 is 1761bp and the protein molecular weight is about 68 KD.
Construction of the plasmid vector of teiR gene from Comamonas testosteroni and overexpression in E. coli
2008, 16(4): 0- |
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The teiR gene was amplified by PCR from the genomic DNA of C. testosteroni. The PCR products were cloned into plasmid pKtac2 (containing tac promoter)and pK18, constructing the recombinant plasmids pKtac2-teiR and pKteiR100.The two plasmids were transformed into Escherichia coli HB101 respectively. The total cell lysate of the host bacteria was extracted to detect the quantity of teiR using ELISA. The results showed that plasmid containing tac promoter (pKtac2-teiR) had a higher transcription level than plasmid pKteiR100. Meanwhile, the recombinant plasmids were respectively co-transformed into E. coli HB101 with plasmid p6 (containing 3a-HSD/CR gene).The total cell lysate of the host bacteria was extracted to detect the quantity of teiR and 3a-HSD/CR using ELISA. The results indicated that teiR could significantly promote the 3a-HSD/CR gene expression; and the teiR expression of pKtac2-teiR was higher than that of pKteiR100 after co-transformation with p6. But interestingly , the expression quantity of 3a-HSD/CR co-transformed with plasmids pKtac2-teiR and p6 was less than that co-transformed with plasmids pKteiR100 and p6.
Optimizing the expression of the subunit fasG of 987P fimbriae of Enterotoxigenic Escherichia coli(ETEC)
He-Feng LUO Jie WANG Zhong-Yong GOU Jian PENG
2008, 16(4): 0- |
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In our experiment, two E. coli expression vectors(pET28a(+)、pGEX-KG) with different translation intiation regions(TIR) and two E. coli expression strains(BL21(DE3)、Rossetta(DE3)) with different tRNA profiles had been used to obtain four bioengineered bacterium of fasG gene. The aim of the experiment was to examine the influence of secondary structures of TIR and the rare codens to the expression of fasG gene. The result of inducing expression of the bioengineered bacteriums showed that the secondary structures of TIR and the existence of rare codens can both influence the expression of target genes, optimizing either could both enhance the expression lever of target gene. The plasmid pKG-fasG had optimized the secondary structures of TIR and Rossetta(DE3) had overcome the rare condons’effects, so the expression lever of pKG-fasG Rossetta(DE3) achieved 16%, add to 100% in comparison with pET-fasG BL21(DE3)(8%). The result of this experiment indicated that the selection of appropriate expression vectors and strains was very important in the expressions of heterologous genes.
Genetic Diversity Analysis of Ralstonia Solanacearum Based on RAMS Marker and ITS Sequence
2008, 16(4): 0- |
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Genetic diversity among 21 strains of Ralstonia solanacearum from different climate zones and different host crop was analyzed by ITS sequence and RAMS methods respectively. Three types of ITS sequence were obtained. Among them, there was only one or two base diversity, and ITS sequence of tested strains was the same to GMI1000 or one or two base difference to GMI1000.The total genomic DNA was then examined by sensitive RAMS technique. The results revealed an obvious diversity of their genomic DNA. Cluster analysis based on the RAMS bands showed that tested isolates could be separated into 3 clusters.The strains come from the same host crop were clustered into one group, and the strains come from the same climate zones were belonged to the same subgroup.It is concluded that genetic diversity of R.solanacearum comes from different host crop and different climate zones, mostly comes from different host crop.
Genetic diversity of Cryphonectria parasitica populations revealed by ISSR analysis
Ke-Rong WANG Li-Qiang XU
2008, 16(4): 0- |
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Genetic diversity and genetic differentiation of Cryphonectria parasitica,which were from China, Japan, Italy and the USA, were analyzed using inter simple sequence repeat (ISSR) molecular technique. Using 11 primers filtrated from total 53 primers, ISSR amplification was conducted in 220 individuals from eleven Cryphonectria parasitica subpopulations which were from four countries, and 174 polymorphic loci were detected from total 177 loci. The percentage of polymorphic loci (P) of the total populations was 98.31%, while that at a subpopulation level averaged 20.43 %. Shannon’s Information index (I) and Nei’s gene diversity (h) of the fungus at the total populations level was 0.1769 and 0.7386, respectively. The mean of I and h at a subpopulation level was 0.0943 and 0.0614, respectively. It indicated that the genetic diversity of C. parasitica between the populations was relatively high. The genetic differentiation coefficient (Gst) among populations was 0.7386. The gene flow (Nm) among populations was low down to 0.1769. The mean of pair-wise genetic distance between eleven C. parasitica subpopulations was 0.2289. These eleven subpopulations could be clustered into three groups using unweighted pair group method with arithmetic mean (UPGMA).
Study on the effect of the activation of immunological cells in the commom carp after stimulation by Lentinan
2008, 16(4): 0- |
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The head kidney macrophages and peripheral blood granulocytes isolated from the head kidney of Cyprinus carpio were purified by means of Percoll gradient density centrifugation and were exposed to Lentinan in vitro. The effects of the polysaccharide on the peripheral blood granulocytes proliferation, respiratory burst of macrophage and IL-1βmRNA expression of macrophages were investigated by using MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) ,NBT(nitroblue tetrazolium) reduction, Griess reagent coloration and Real time PCR respectively, to evaluate the immunostimulant of the polysaccharide and compare its immune activities with the control group. Results showed that Lentinan stimulated markedly the proliferation of the peripheral blood granulocytes and could induce superoxide anion in macrophages markedly. Lentinan had no conspicuous effect on nitrogen burst activity of macrophages, moreover it inhibited the nitrogen burst activity at higher doses. The polysaccharide stimulated IL-1βexpression in carp head kidney . It is suggested that Lentinan could modulate the immune response of Cyprinus carpio.
Cloning and sequence analysis of heteromeric ACCase subunit encoding gene accd from four close-related species of Brassica
2008, 16(4): 0- |
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The cDNA fragments of accd gene encoding for subunit β-carboxyltransferase of heteromeric Acetyl-CoA carboxylase (ACCase) were amplified from young leaves cDNA of Brassica napus, Brassica oleracea, Brassica juncea and Brassica rapa by means of RT-PCR. The lengths of the amplified accd genes were 1470bp, 1470bp, 1464bp and 1464bp respectively. Sequence analysis showed that the cDNA sequences of the four accd genes shared high homology. The amino acid sequences deduced from the four accd genes contained the similar zinc finger structure and five same motifs on C-terminal regions, among which motif I (GSMGSVVG) and motif II (PLIIVCASGGARMQE) were present in β-carboxyltransferases of all plant species and E.coli. Southern blotting demonstrated that there was only a single copy of accd gene in genomes of the four close-related species of Brassica.
Establishment of a McAb-based Competitive ELISA for the detection of antibodies against Pasteurella multocida toxin
2008, 16(4): 0- |
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Abstract:Monoclonal antibodies against Pasteurella multocida toxin(PMT) were produced by using E.coli expressed recombinant PMT proteins as immunogens.on the basis of recombinant protein expressed by E.coli and HRP-labled monoclonal antibody against Pasteurella multocida toxin,a competitive ELISA was developed to detect antibodies against.Pasteurella multocida toxin.Optimium conditions of the ELISA Were developed as following:the concentration of recombinant for coating ELISA plates is 223 ng /mL;the best dilution of serum to be tested was 1:2; the working titer for HRP-labelled monoclonal antibody was 1:3200;>50% of inhibition rate was the positive judging standard.A total of 82 serum samples were detected in parallel by both the competitive ELISA and in vitro neutralization assay.40.2% of them were detected as positive by competitive ELISA,while 36.6% of them were positive by in vitro neutralization assay.The coincidence rate of the two assays is 91.5%.These results showed that the cELISA has the advantage of higher sensitivity,specificity,reproducibility and stability.It would be very useful in diagnosis,surveillance of PMT antibodies and epidemiological survey.
An Indirect ELISA for Detection of Antibody against Akabane Virus Using Recombinant Truncated Glycoprotein G1 Expressed in Escherichia coli
2008, 16(4): 0- |
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To obtain a recombinant Glycoprotein G1 of Akabane virus for diagnostic purpose, the gene fragment coding the high antigenic domain of AKAV G1 protein, was amplified from the AKAV genome by RT-PCR after analyzing glycoprotein G1 with bio-software. The amplified product was cloned into pMD18-T vector and sequenced. Then the gene fragment was subcloned into pET-28a (+) vector directionally. The recombinant plasmid was transformed into E. coli BL21 (DE3). SDS-PAGE and Western-blot analysis indicated that the fusion protein was insoluble and approximately 44 ku in molecular weight and had immunological activity. After purified by affinity chromatography, the concentration of purified protein was 2mg/ml and the purity was 92.6%. An indirect enzyme linked immunosorbent assay (ELISA) was developed using the purified recombinant protein. The optimal reaction conditions of ELISA were determined: the coating concentration of the purified recombinant protein was 20g/ ml; the dilution fold of the serum samples was 1:200. There was no cross reaction with positive sera of other six infectious diseases. Bovine serum samples from Yun Nan (77) and Inner Mongolia (70) had been detected by serum neutralizing (SN) test before were detected by this ELISA. In accordance with SN test, the positive threshold of the ELISA was determined as 0.493 and 0.488 in two districts respectively. Comparing to SN test, the speciality of the ELISA was 73% and 86.9% respectively. The agreement ratio between the two methods was 80%(52/65) and 85.9%(55/64) respectively.
The CDS Cloning of HCRTR1 Gene with Over-lap PCR from Qinchuan Cattle and its Expression in E.coli
2008, 16(4): 0- |
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With the Overlap-PCR methods, the 1 278 bp cds sequences of HCRTR1 gene in Qinchuan cattle was got. The fragments were cloned into the pMD18-T vector and the positive clone plasmids was sequenced. Results showed that the cloned fragments had 98% homology with the sequence of GenBank after being blasted. Digested the plasmid with BamHⅠand HindⅢ and the 1 278 bp was obtained. The obtained fragment was ligated with the longer fragment digested with the same restriction enzyme from the expression vector, pET32 a(+). The positive recombinated plasmid was transformed Bl21(DE3) and induced with IPTG. But the HCRTR1 gene cannot be expressed by E.coli. And the tranmembrane structure may explain why.
Molecular cloning and characteristics of cDNA encoding sheep C3d
2008, 16(4): 0- |
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In this study, we first molecularly cloned sheep C3d from RNA of the sheep liver, and compared with the C3d gene of other animals available in GenBank at nucleotide and amino acid leves. The 909bp cDNA of sheep C3d f encodes a polypeptide of 303 amino acids consisting of 1 potential Glycosaminoglycan attachment sites(SG.G), 2 Protein kinase C phosphorylation sites([ST].[RK]) , 1Tyrosine kinase phosphorylation site, 2 N-myristoylation sites and 1 Casein kinase II phosphorylation site. The amino acid sequence similarity of C3d between human, bovine, goat, boar, golden hamste,rmouse,brown rat,Macropus, hare,Gallus is 80.4% ,94.7% ,96.9% ,82.8% ,81.1% ,80.6% ,80.2%,71.0% ,75.2% and60.3% respectively..Helix and angles turns out alternately in Sheep C3d second structure, which helix accounted for 55.12%, strand to 2.31%and coil for the 42.57% of such a structure conducive to the performance of barrel-shaped structure is formed. ESyPred3D through homology modeling forecast sheep C3d three known structures C3d with the same people also formed by nuclear heart and the outer form of the performance of barrel-shaped molecular structure. The experiment not only for livestock C3d in the immunological basis to provide experimental evidence, but also for the construction of high immunogenicity of vaccine gene lay the foundation for the new technology.
Cloning and differently expression analysis of porcine MYL4 gene in embryo skeletal muscle
Xiao xiong WU
2008, 16(4): 0- |
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The cDNA sequences of porcine MYL4 gene were cloned, and verified by bioinformatics method. The cDNA fragment was 1105 bp with an entire open reading frame (ORF), encoding 197 amino acids. The analysis of sequence revealed that the porcine MYL4 gene was highly homologous with that of other species such as canis lupus familiaris (98.4%), Homo sapiens (95.9%),chimpanzee (95.4%) and rhesus monkey(95.4%) in amino acid. The protein encoded by porcine MYL4 gene had conserved domain ‘EFH’ and ‘FRQ1’. The expression analysis of MYL4 gene was carried out in the fetus skeletal muscle (at 33, 65 and 90 days post coitus, dpc) by real-time quantitative PCR. The results indicated that MYL4 gene was expressed with wave model in both Tongcheng and Landrace pigs. There was significant difference at 65 and 90 dpc, but no significant difference at 33 dpc between Tongcheng and Landrace.
Cloning of 3′-end full length of cDNA encoding for MT-I/Ⅲ and analyzing of the sequence in Yak
Bin LIU Jian-ping WU Bin-yun MA Feng-qin GAO
2008, 16(4): 0- |
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Abstract: In the present study, the domestic yak (B.Grunniens) metallothionein-I and Ⅲ cDNA 3′-end full length sequences (331bp, 378bp) were amplified and cloned by RT-PCR and 3′-RACE using the primer YMT-ⅠSP, YMT-ⅢSP and M13 Primers M4, in which included MT-I and MT-Ⅲ cDNA coding sequences (183bp, 207bp) and AATAAA and Poly(A)in the end. The cDNA sequences of MT-I and MT-Ⅲ were subjected to Blastn searching in CBI and the results showed that the nucleotide sequences of yak MT-I and MT-Ⅲ between different species mammal are comparatively conservative. The yak cDNA sequence coding for yak MT-I protein is composed of 61 amino acids, including 20 Cysteine. There are lots of conserved tripetide, such as C-X-C, C-C-X-C-C, C-X-X-C, etc.. The yak cDNA sequence coding for yak MT-Ⅲ protein is composed of 68 amino acids, including 19 Cysteine, which has the same conserved tripetide with MT-Ⅰ, and MT-Ⅲ has also its own particular conserved tripetide, such as T, CPCP,GEGAEA, etc.. They are all comparatively conservative in their evolution.
The extrusion on bovine oocytes induced with demecolcine
2008, 16(4): 0- |
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This study aimed to assess some factors effecting the formation of nuclear extrusion on demecolcine-induced bovine oocytes, including oocyes with or without the first polar body, different demecolcine treatment concentration and duration. The result demonstrated: treated with 0.5 μg/mL demecolcine, oocytes induced for 1 h (76.00%) gained higher rate of extrusion than those for 0.5 h (61.18%)、1.5 h (64.10%) and 2 h (63.46%). And in 1 h demecolcine-treatment, 0.4 μg/mL and 0.5 μg/mL demecolcine led to the higher rate of extrusion (75.24% and 77.31% respectively) compared to the 0.3 μg/mL (46.54%) and 0.6 μg/mL group (60.66%). In addition, there was no significant difference on the extrusion rate between oocytes with (83.24%) or without (77.54%) first polar body. To sum up, bovine oocytes induced with 0.4 μg/mL demecolcine for 1h can gain the higher rate of extrusion. And demecolcine can assist enucleation effectively in Nuclear Transfer.
The cDNA Cloning ,Sequencing and Tissue Distribution of Chicken β-Defensin Gallinacin Gal-1~Gal-13 Genes in Hu-xu Chicken
2008, 16(4): 0- |
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With specific primers,13 chicken β-defensin Gallinacin-1(Gal-1)~Gallinacin-13(Gal-13) cDNA fragments, which from Hu-xu chicken strains, were amplified by reverse transcription-polymerase chain reaction(RT-PCR). And they all coded the full-length open reading frame,except Gal-3. After sequenced, 13 cDNA fragments were identified and submited to Genbank, and received 13 accession numbers: DQ858311~ DQ858323. It was found that the 13 chicken β-defensin genes had similarities ranging from 96.5%to 100% at amino acid level.When compared with β-defensin genes sequence in Genbank. Analysis by Clustalx revealed that Gal-1、Gal -1(a) and Gal -3 lie at the same embranchment; Gal-4, Gal-5, and Gal-8 lie at the same other one; and Gal-2 ,Gal-12 and Gal-13 lie at the same; Gal-6 and 7 lie at the same ;Gal-11 is the embranchment independently. Differential mRNA expression of genes in different tissues by RT-PCR had been shown that Gal-1, Gal-2, Gal-4,Gal-5,Gal-6,Gal-7 and Gal-10 are widely expressed in tissues; Gal-3,Gal-8,Gal-11,Gal-12 and Gal-13 have tissue specific expression.
Isolation and characterization of microsatellites
2008, 16(4): 0- |
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We report here the isolation and characterization of microsatellites, in Pelteobagrus fulvidraco using an enrichment protocol. The size-fractionated genomic library was screened with (AC)8、(CT)8、(AT)7、(GATA)8 and (GATT)7 oligonucleotide probes. A total of 38 clones were identified as having the microsatellite repeats, and specific 22 primer pairs were designed for 38 microsatellite loci. Of the 22 primer pairs, 18 amplified polymorphic loci in Pelteobagrus fulvidraco . At these polymorphic loci, the number of alleles per locus varied from 2 to 8(the average 3.78), the observed heterozygosities ranged from 0.2225 to 0.8101(the average 0.5755), and the polymorphism information content ranged from 0.3425 to 0.7900(the average0.5336). These 18 primer pairs were examined also in Pelteobagrus species and Siluriformes species.16 of the 18 primer pairs amplified the loci in the Pelteobagrus species,13 of the 18 primer pairs amplified the loci in the Siluriformes species. These results indicate that the microsatellites isolated in this study were highly informative and could be useful tools for genetic analysis in the Pelteobagrus species and the Siluriformes species.
Microsatellite Enrichment by Magnetic Beads in Erythroculter ilishaeformis
2008, 16(4): 0- |
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According to the hadro-affinity principle of biotin and streptavidin, Microsatellite DNAs were isolated from Erythroculter ilishaeformis genome by combining biotin capture method and radioactive labeling hybridization. In this experiment, 613 (38.31%)positive colonies were obtained through 1400 colonies. Sequencing of 288 positive colonies confirmed that it contained 280 microsatellite loci. From these sequences,204 repeat motifs (about 72.86%) were perfect,49 repeat motifs (about 17.5%) were imperfect, 27 repeat motifs (about 9.64%) were compound. 243 pairs of primers were designed with the software Primer 3.0 .In addition, 50 pairs of primers were composed and screened. 30 pairs were used successfully to amplify special fragemnts, among which 12 pairs were polymorphism within species
Auxin regulation in the interaction between Rice stripe virus and rice
2008, 16(4): 0- |
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The relative expression level of YUCAA1 gene and the amount of endogenous IAA both in RSV infected rice plants and rice suspension cells were investigated by real-time RT-PCR and high performance liquid chromatography respectively. In rice suspension cells, RSV infection resulted that the expression of YUCAA1 gene and the amount of endogenous IAA were increased 2.98 and 2.66 times respectively. Whereas in rice plants, RSV infection leaded to decrease 70% and 75% on the expression of YUCAA1 and the amount of endogenous IAA respectively. These results indicated that RSV infection could induce auxin signal transduction in rice. At the same time, the expression of RSV CP was increased 2.9 times in rice plant with KPSC buffer treatment after depleting endogenous auxins, and was decreased 45% with 30 μM IAA treatment. These suggested that auxin could regulate RSV replication in rice plant.
Construction and Expression of a Brassica napus Myrosinase in Escherichia coli
2008, 16(4): 0- |
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The myrosinase gene was amplified from total RNA by RT-PCR and inserted into Escherichia coli expression vector pGEX-4T-1.The result of sequencing was EF583560 in GenBank.The amino acid sequence showed that the homology was 99% comparing with a length of published amino acid sequence of Q00326.The recombinant protein was highly expressed at 91KDa by IPTG induced in Escherichia coli.
研究简报
Gene Cloning and Expression of Porcine lnterleukin-18 Mature Protein in Es-cherichiacoli
2008, 16(4): 0- |
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Polymorphism of Prolactin Receptor Gene and Its Relationship with Prolificacy of Jining Grey Goats
2008, 16(4): 0- |
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Prolactin receptor(PRLR)gene was studied as a candidate gene for the prolificacy of Jining Grey goats. Eight pairs of primers were designed to detect single nucleotide polymorphisms of exon 3 to exon 9 and intron 9 of PRLR gene in both high fecundity breed (Jining Grey goat) and low fecundity breeds (Liaoning Cashmere goat and Angora goat) by PCR-SSCP. Only the products amplified by primer P8 amplifying the intron 9 displayed polymorphisms. For primer P8, only two genotypes (AA and AB) were detected in three goat breeds. Sequencing revealed one mutation (209A→G) of PRLR gene in the genotype AB in comparison to the genotype AA, and this mutation was situated in intron 9 of PRLR gene. The difference of the least squares means for litter size between AA and AB was non-significant (P>0.05) in Jining Grey goats. These results preliminarily showed that these detected loci of PRLR gene had no significant effect on prolificacy of Jining Grey goats.
Cloning and Sequencing of Chitosanase gene
MaDi
2008, 16(4): 0- |
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The research used one pair to direct the thing to clone the entire gene sequence of Csn from Bacillus megaterium BS-0409 which the size is 516bp, retrieval analysis online with the BLAST procedure in the NCBI, the result indicate that bacillus homology is 96%. At the same time, compare with Csn code gene amino acid sequence other 13 kind of different bacterium uses DNAMAN software, it demonstrated the quite homology between the different bacterium with many kinds of bacillus.
专家论坛
Potential Commercialization of Genetically Modified Rice in China: Key Questions for Environmental Biosafety Assessments
2008, 16(4): 0- |
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The commercialization of genetically modified (GM) rice may arouse environmental biosafety concerns, including the impacts of transgenes on non-target organisms and biodiversity, transgene escape and its related ecological consequences, and transgenic influences on soil micro-fauna form extensive cultivation of GM rice. Science-based environmental biosafety assessments of GM rice will facilitate its safe and sustainable application in agriculture eco-systems. Based on the actual eco-systems and cultivation styles of rice, as well as the types of GM rice varieties that would be possibly commercially released in China, this paper presents a rational analysis of potential environmental biosafety consequences caused by the extensive commercial cultivation of the GM rice. Based on the principal of any risk assessment (risk% = hazards × exposure), the author particularly analyzed the direct and indirect influences of GM rice resistant to insects, diseases, and herbicides. Hopefully, the analysis based on the rational thinking will provide useful information for decision-making on GM rice commercialization and the relevant environmental biosafety assessments.
生物技术性动态
Progress in DetectionTechnique for Genetically Modified Organisms
2008, 16(4): 0- |
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The detection technique for Genetically Modified Organisms(GMO), including qualitative PCR, quantitative PCR, Enzyne-Linked Immunosorbent Assay and other detection technique, are systematically illustrated. The application progress of GMO in species-special detection, endogenous gene, standard substance and restrain factor influencing detection are reviewed, and the present problems and development prospects of detection technique for GMO are pointed out
Research Progress on Transcription Factor DREB
2008, 16(4): 0- |
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Abstract: Biotic and abiotic stresses negatively influence the growth and development of plants and restrict the development of agriculture and forestry. Many transcription factors involve in the expression regulation of stress related genes by stress signal transduction,of which, DREB is one of the most important transcription factors. In this paper, the research progress on DREB in recent years was reviewed. The researches on DREB provided the important information to understand the molecular mechanism of plant stress resistance and the opportunity to develop stress resistant plants.
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