Based on the nucleotide sequence of porcine reproductive and respiratory syndrome virus (PRRSV) CH 1a strain, a pair of primers was designed. The GP3 gene of PRRSV HB 1 strain was cloned by RT PCR. The sequences analy...Based on the nucleotide sequence of porcine reproductive and respiratory syndrome virus (PRRSV) CH 1a strain, a pair of primers was designed. The GP3 gene of PRRSV HB 1 strain was cloned by RT PCR. The sequences analysis showed that the GP3 genes of HB 1 and CH 1a strain had 91% and 89% homology, at the levels of the nucleotide and amino acid, respectively. The GP3 gene was digested by Bam HⅠ and Eco RⅠ, and the fragment was inserted into the same sites of the universal vector pPgG uni. A recombinant virus transfer vector pPgG GP3 expressing PRRSV GP3 gene was constructed. The transfer vector pPgG GP3 digested by Kpn Ⅰ was co transfected PK 15 cells with the PRV TK -/gG -/LacZ + genomic DNA digested by Eco RⅠ using liposome method. The recombinant virus was purified by the phage test and PCR amplification. The expression of GP3 gene was indicated by Western blot analysis using anti sera. A recombinant PRV virus expressing PRRSV GP3 gene was obtained. The recombinant virus is very useful in the research of the genetic engineering vaccine against pseudorabies virus and PRRS virus.展开更多
目的探讨川北地区汉族人群调节性T细胞(Treg)转录因子叉头蛋白p3(Foxp3)基因rs3761548多态性与经典1型糖尿病(T1DM)的相关性。方法回顾性分析2012年4月~2020年9月我院内分泌科收治的180例经典T1DM患者(T1DM组)及同期本地186名无血缘关...目的探讨川北地区汉族人群调节性T细胞(Treg)转录因子叉头蛋白p3(Foxp3)基因rs3761548多态性与经典1型糖尿病(T1DM)的相关性。方法回顾性分析2012年4月~2020年9月我院内分泌科收治的180例经典T1DM患者(T1DM组)及同期本地186名无血缘关系的我院健康体检者(对照组)为研究对象。利用聚合酶链反应和质谱法对Foxp3基因rs3761548位点进行基因分型。酶法检测空腹血糖(FPG)、餐后2小时血糖(2 h PG)、血清甘油三酯(TG)、总胆固醇(TC)及高密度脂蛋白胆固醇(HDL-C);化学发光法检测空腹C肽(FCP)、餐后2小时C肽(2 h CP);液相色谱法检测糖化血蛋白A_(1c)(HbA_(1c))。结果Treg细胞转录因子Foxp3基因rs3761548多态性位点等位基因频率和基因型频率分布均符合Hardy-Weinberg平衡定律。rs3761548多态位点A等位基因频率为18.7%。T1DM组和对照组之间rs3761548多态位点A等位基因的频率差异无统计学意义(19.2%vs 18.3%,P>0.05)。T1DM患者rs3761548多态位点A等位基因携带者的LDL-C水平更高,而FCP、2 h CP水平更低(P<0.05)。结论Foxp3基因rs3761548多态性A等位基因与川北地区汉族T1DM患者LDL-C水平及FCP、2 h CP水平密切相关。展开更多
文摘Based on the nucleotide sequence of porcine reproductive and respiratory syndrome virus (PRRSV) CH 1a strain, a pair of primers was designed. The GP3 gene of PRRSV HB 1 strain was cloned by RT PCR. The sequences analysis showed that the GP3 genes of HB 1 and CH 1a strain had 91% and 89% homology, at the levels of the nucleotide and amino acid, respectively. The GP3 gene was digested by Bam HⅠ and Eco RⅠ, and the fragment was inserted into the same sites of the universal vector pPgG uni. A recombinant virus transfer vector pPgG GP3 expressing PRRSV GP3 gene was constructed. The transfer vector pPgG GP3 digested by Kpn Ⅰ was co transfected PK 15 cells with the PRV TK -/gG -/LacZ + genomic DNA digested by Eco RⅠ using liposome method. The recombinant virus was purified by the phage test and PCR amplification. The expression of GP3 gene was indicated by Western blot analysis using anti sera. A recombinant PRV virus expressing PRRSV GP3 gene was obtained. The recombinant virus is very useful in the research of the genetic engineering vaccine against pseudorabies virus and PRRS virus.
文摘目的探讨川北地区汉族人群调节性T细胞(Treg)转录因子叉头蛋白p3(Foxp3)基因rs3761548多态性与经典1型糖尿病(T1DM)的相关性。方法回顾性分析2012年4月~2020年9月我院内分泌科收治的180例经典T1DM患者(T1DM组)及同期本地186名无血缘关系的我院健康体检者(对照组)为研究对象。利用聚合酶链反应和质谱法对Foxp3基因rs3761548位点进行基因分型。酶法检测空腹血糖(FPG)、餐后2小时血糖(2 h PG)、血清甘油三酯(TG)、总胆固醇(TC)及高密度脂蛋白胆固醇(HDL-C);化学发光法检测空腹C肽(FCP)、餐后2小时C肽(2 h CP);液相色谱法检测糖化血蛋白A_(1c)(HbA_(1c))。结果Treg细胞转录因子Foxp3基因rs3761548多态性位点等位基因频率和基因型频率分布均符合Hardy-Weinberg平衡定律。rs3761548多态位点A等位基因频率为18.7%。T1DM组和对照组之间rs3761548多态位点A等位基因的频率差异无统计学意义(19.2%vs 18.3%,P>0.05)。T1DM患者rs3761548多态位点A等位基因携带者的LDL-C水平更高,而FCP、2 h CP水平更低(P<0.05)。结论Foxp3基因rs3761548多态性A等位基因与川北地区汉族T1DM患者LDL-C水平及FCP、2 h CP水平密切相关。
