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目的 研究猴免疫缺陷病毒 (SIV)外膜糖蛋白 (envGP)基因片段与病毒复制功能的关系。方法 利用两病毒株共有的内切酶位点 ,将非致病性SIVmac 142株envGP区的DNA片段与致病性SIVmac 2 39株的对应区域进行置换 ,构成多个重组体。RFLP和部分序列测序确定后 ,用等量突变病毒 (3μg)转染CD4+ T淋巴细胞CEM× 174细胞系 ,用ELISA监测培养液中SIV核心蛋白P2 7水平的变化 ,以判定重组病毒的复制能力。结果 与SIVmac 2 39株相比 ,SIVmac 2 39envGP重组体 (SIV mac142 /2 39envGP/142 )仍保持高度的复制活性 ,SIVmac2 39gp41(SIVmac142 /2 39gp41/142 )或SIV mac2 39N gp41(SIVmac142 /2 39N gp41/142 )的复制能力明显降低 ,而SIVmac2 39C gp41(SIVmac142 /2 39C gp41/142 )重组体无复制活性。结论 envGP基因是SIVmac2 39株复制能力或毒力的重要调节因素。
Objective To study the relationship between env gene fragment and virus replication in simian immunodeficiency virus (SIV). Methods Using the restriction endonuclease sites shared by the two strains, the envGP region of the non-pathogenic SIVmac 142 strain was replaced with the corresponding region of the pathogenic SIVmac 2 39 strain to construct multiple recombinants. After RFLP and partial sequencing were confirmed, the CD4 + T lymphocyte CEM × 174 cell line was transfected with the same amount of mutant virus (3 μg), and the changes of SIV core protein P2 7 in the culture solution were monitored by ELISA to determine the replication ability of the recombinant virus . As a result, the SIVmac239envGP recombinant (SIV mac142 / 2 39envGP / 142) retained a high degree of replicative activity compared to SIVmac 2 39 strain, and SIVmac2 39gp41 (SIVmac142 / 2 39gp41 / 142) or SIV mac2 39N gp41 (SIVmac142 / 2 39N gp41 / 142), while the replication of SIVmac2 39C gp41 (SIVmac142 / 2 39C gp41 / 142) did not replicate. Conclusion The envGP gene is an important regulator of replication or virulence of SIVmac2 39 strain.