[Objective] The study aimed to clone RPO30 gene from Sheeppox virus （SPPV） and predict the structure and function of the sequence. [Method] RPO30 gene of SPPV was cloned with PCR, linked into pMD18-T simple vector a...[Objective] The study aimed to clone RPO30 gene from Sheeppox virus （SPPV） and predict the structure and function of the sequence. [Method] RPO30 gene of SPPV was cloned with PCR, linked into pMD18-T simple vector and then transformed into E. coli DH5a. In blue-white screen, the white colonies were selected to prepare plasmids. The positive plasmids were selected by double digestion and PCR, and then sequenced. Finally, the structure and function of the sequence obtained were predicted by bioinformatics methods. [Results] The RPO30 gene was successfully obtained; its ORF was 585 bp, encoding 193 amino acids and containing a recognition site for Hind III. Moreover, the SPPV RPO30 gene shared different homologies with the RPO30 gene sequences of other pox virus strains from GenBank database. Further analysis by biological software showed that in RPO30 protein, amino acids 4-12, 18-26, 50- 61, 68- 92 and 176-190 had a high possibility to form the active center, and acting to these regions was likely to inactivate the enzyme encoded by the sequence, thus to inhibit viral replication efficiently. [Conclusion] This study will lay foundation for further study on the structure and function of RPO30.展开更多
The genetic polymorphism of SLA-DQA gene was investigated by PCR-RFLP. The effects of SLA.DQA gene on birth weight and 30-day-old weight were analyzed in 250 individuals of FI hybrid pig （ Heilongjiang wild boar x Be...The genetic polymorphism of SLA-DQA gene was investigated by PCR-RFLP. The effects of SLA.DQA gene on birth weight and 30-day-old weight were analyzed in 250 individuals of FI hybrid pig （ Heilongjiang wild boar x Beijing Black sow）. The analysis revealed four poly- morphic loci in the SLA-DQA gene, but only the Pvu II locus in exon 2 of SLA-DQA gene （ BB genotype） greatly increased the 30-day-old weight of piglets.展开更多
The gene encoding the major surface antigen(P30) of Toxoplasrna gondii was cloned into a transfer plasmid vector pSXIVVI￣+X3,then the recombinant plasmid pSXIVVI￣+X3-P30 DNA and the parent virus TnNPV DNA were used ...The gene encoding the major surface antigen(P30) of Toxoplasrna gondii was cloned into a transfer plasmid vector pSXIVVI￣+X3,then the recombinant plasmid pSXIVVI￣+X3-P30 DNA and the parent virus TnNPV DNA were used to cotransfect the cultured Spodoptera f展开更多
目的探讨系统性红斑狼疮(system ic lupus erythem atosus,SLE)患者CD4+T细胞mRNA中干扰素诱导蛋白30基因(IP-30)的表达水平与SLE疾病活动度的相关性。方法收集23例SLE患者和10例正常对照人群的临床资料。取外周血用免疫磁珠法分离出CD...目的探讨系统性红斑狼疮(system ic lupus erythem atosus,SLE)患者CD4+T细胞mRNA中干扰素诱导蛋白30基因(IP-30)的表达水平与SLE疾病活动度的相关性。方法收集23例SLE患者和10例正常对照人群的临床资料。取外周血用免疫磁珠法分离出CD4+T细胞,抽提RNA并逆转录合成cDNA,运用GLG I方法从LongSAGE标签库中筛选出IP-30,比较该基因在系统性红斑狼疮疾病活动指数(SLEDAI)不同的SLE患者中表达的差异。结果①SLE患者组IP-30的表达量显著高于正常对照(P=0.01,P<0.05)。SLEDAI≥10组和SLEDAI<10组其IP-30的表达量显著高于正常对照(P分别为0.01和0.047),但两组间比较无显著差别(P=0.149)。②SLE有狼疮肾炎组和SLE无狼疮肾炎组相比IP-30表达量有差异(P<0.05)。③IP-30的表达量随SLE的活动水平升高而明显增加,与SLEDAI具有显著的正相关(r=0.830,P<0.01),与补体C3和外周血白细胞数成负相关(r=-0.517,r=-0.424,P<0.05)。结论CD4+T细胞IP-30水平表达升高提示IP-30可能参与SLE的发病,并对SLE活动度的判断和狼疮肾炎的诊断有一定的意义。展开更多
基金Supported by the National Natural Science Foundation of China(31001056)the National Natural Science Foundation of China(31101802)+1 种基金Major Program for New Transgenic Organism Verities Breeding of Ministry of Agriculture of China(2009ZX08008-010B)Key Science and Technology Foundation of Gansu Province(092NKDA032)~~
文摘[Objective] The study aimed to clone RPO30 gene from Sheeppox virus （SPPV） and predict the structure and function of the sequence. [Method] RPO30 gene of SPPV was cloned with PCR, linked into pMD18-T simple vector and then transformed into E. coli DH5a. In blue-white screen, the white colonies were selected to prepare plasmids. The positive plasmids were selected by double digestion and PCR, and then sequenced. Finally, the structure and function of the sequence obtained were predicted by bioinformatics methods. [Results] The RPO30 gene was successfully obtained; its ORF was 585 bp, encoding 193 amino acids and containing a recognition site for Hind III. Moreover, the SPPV RPO30 gene shared different homologies with the RPO30 gene sequences of other pox virus strains from GenBank database. Further analysis by biological software showed that in RPO30 protein, amino acids 4-12, 18-26, 50- 61, 68- 92 and 176-190 had a high possibility to form the active center, and acting to these regions was likely to inactivate the enzyme encoded by the sequence, thus to inhibit viral replication efficiently. [Conclusion] This study will lay foundation for further study on the structure and function of RPO30.
基金supported by the Key Projects in the National Science & Technology Pillar Program in the 11th Five Years (2008BADB2B02)
文摘The genetic polymorphism of SLA-DQA gene was investigated by PCR-RFLP. The effects of SLA.DQA gene on birth weight and 30-day-old weight were analyzed in 250 individuals of FI hybrid pig （ Heilongjiang wild boar x Beijing Black sow）. The analysis revealed four poly- morphic loci in the SLA-DQA gene, but only the Pvu II locus in exon 2 of SLA-DQA gene （ BB genotype） greatly increased the 30-day-old weight of piglets.
文摘The gene encoding the major surface antigen(P30) of Toxoplasrna gondii was cloned into a transfer plasmid vector pSXIVVI￣+X3,then the recombinant plasmid pSXIVVI￣+X3-P30 DNA and the parent virus TnNPV DNA were used to cotransfect the cultured Spodoptera f
