含抗HIV广谱中和抗体2F5、4E10靶基因载体的构建及鉴定

目的:构建含HIV gp120,gp41序列中广谱中和抗体2F5,4E10作用靶基因的载体并进行鉴定,为后期重组载体表达产物诱导产生中和抗体及抗HIV亚单位疫苗的研究奠定基础.方法:根据NCBI中HIV gp120,gp41基因序列中可与2F5、4E10结合的区域设计引物并进行PCR反应,将PCR得到的目的片段插入到载体pET28a中,对重组载体进行PCR鉴定、酶切鉴定及DNA测序.结果:PCR鉴定、酶切鉴定及DNA测序结果证实重组载体构建成功.结论:成功构建了含HIV gp120,gp41序列中广谱中和抗体2F5,4E10作用靶基因的载体.

Construction and Identification of Antibodies 2F5 and 4E10 Target Gene Vectors with the Neutralization of Anti-HIV Broad Spectrum

Objective Antibodies 2F5 and 4E10 target gene vectors with the neutralization of anti-HIV broad spectrum were constructed and identified.This will certainly lay the foundation for future research of neutralizing antibodies from recombinant vector products in expression and induction and anti-HIV subunit vaccine.Method Primers were designed and PCR was conducted according to the binding region of the HIV gp120,gp41 gene sequences in NCBI and the 2F5,4E10.Then the target fragments from PCR were inserted into the vector pET28a.In the end,PCR,restriction enzyme digestion and DNA sequencing were performed for the recombinant vectors.The results: PCR,restriction enzyme digestion and DNA sequencing confirmed that the recombinant vector were successfully constructed.The conclusion: The construction of vectors of broad-spectrum neutralizing antibodies 2F5,4E10 target genes containing HIV gp120,gp41 gene sequences was successfully conducted in this paper.