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目的 构建抗HIV 1gp120单链抗体基因,为进一步用于HIV 1感染的诊断和治疗奠定基础。方法 利用 RT PCR法,从抗HIV 1gp120单克隆抗体杂交瘤细胞扩增得到抗体轻链和重链的可变区基因,经重叠延伸反应, 在体外随机合成单链抗体基因(ScFv),并克隆到pGEM Easy T载体中,经测序及Blast同源性分析。结果 该 ScFV基因全长为666bp,为VH Linker VL结构,VH基因为396bp,编码132个氨基酸;Linker为(Gly4Ser)3短 肽;VL基因为270bp,编码90个氨基酸。VH基因与小鼠IgG2a重链可变区的同源性达95%,VL基因与小鼠免 疫球蛋白κ轻链可变区的同源性达98%。结论 成功构建了抗HIV 1gp120单链抗体基因。
Objective To construct the anti-HIV 1gp120 single-chain antibody gene and lay a foundation for the further diagnosis and treatment of HIV-1 infection. Methods The variable region genes of the light chain and the heavy chain of the antibody were amplified by RT PCR from the monoclonal antibody against HIV 1 gp120. The scFv gene was cloned in vitro by overlap extension reaction. Into pGEM Easy T vector, sequenced and Blast homology analysis. Results The full-length ScFV gene was 666 bp in length. It was a VH Linker VL. The VH gene was 396 bp in size and encoded 132 amino acids. Linker was a (Gly4Ser) 3 short peptide. The VL gene was 270 bp in size and encoded 90 amino acids. The homology between VH gene and mouse IgG2a heavy chain variable region was 95%, and the homology between VL gene and mouse immunoglobulin κ light chain variable region was 98%. Conclusion The anti-HIV 1gp120 single-chain antibody gene was successfully constructed.