mB7-1重组真核载体的构建及表达

目的构建mB7-1真核表达载体并检测其在B16细胞中的瞬时表达情况。方法提取小鼠脾细胞mRNA,PCR法获得mB7-1片段,与pcDNA3连接,并进行酶切鉴定及测序。重组质粒经脂质体介导转染鼠B16细胞,检测基因表达情况。结果完成mB7-1-pcDNA3重组载体连接及鉴定,基因测序与Genebank一致,并转染至B16细胞,检测到基因表达。结论成功构建mB7-1-pcDNA3真核表达载体,获得mB7-1表达细胞系,为研究其抗瘤效应奠定了基础。

The foundation and expression of recombinated vector mB7-1

Objective To costruct the mB7-1-pcDNA3 recombinated vector and observe its expression in B16 cells.Method mB7-1 cDNA was cloned by RT-PCR from BALB/C mouse splenic cells and inserted into pcDNA3 to construct an eukaryotic expression vector（mB7-1-pcDNA3）,which identified by digestion（with EcoRⅠand XhoⅠ） and genetic sequencing.The mB7-1-pcDNA3 plasmid was transfected into B16 cells by liposomes,and detected by RT-PCR.Results Recombinated vector mB7-1-pcDNA3 was obtained and the expression of mB71 was observed in B16 cells.Conclusion The successful construction and expression of mB7-1 pcDNA3 has laid foudation for the further study on tumour immunotherapy.