重组质粒pcDNA3.1-CD44v5的构建及意义

Construction of recombinant plasmid of pcDNA3.1-CD44v5 and its significance

目的:观察构建含人的肿瘤侵袭和转移的特异性抗原CD44v5编码基因的真核表达的重组载体,为进一步研究CD44v5修饰的DC融合瘤苗的免疫治疗作用及机制提供实验依据及理论基础。方法:以pcDNA3.1作为真核表达载体,选择CD44v5编码基因进行RT-PCR扩增,构建重组表达质粒,进行酶切鉴定和测序分析。结果:经酶切鉴定和测序分析表明,插入的目的基因片段为CD44v5的编码基因,由129bp组成,与GeneBank中登录的cDNA相比较,同源性高达100%,且方向正确。结论:成功构建了pcDNA3.1-CD44v5真核表达的重组载体,为下一步构建和研究以CD44v5修饰的DC融合瘤苗抗肿瘤作用及其机制奠定了基础。

Objective：To construct the eukaryon gene expression vector for CD44v5.Methods：CD44v5 gene was amplified by RTPCR and cloned into plasmid pcDNA3.1 to form the recombinant plasmid pcDNA3.1-CD44v5,which was then indentified by the technol-ogy of restriction digest and nucleotides sequencing.Results：The restriction digest and sequence analysis demonstrated that this 129bp of CD44v5 gene was the same as that published on GeneBank,and it was inserted into the vector accurately.Conclusion：A new plasmid of pBud-CD44v5 has been successfully established.This study is the basic foundation for our next experiment on studying the anti-tumor effect and mechanism of fusion hybrid of DCs expression CD44v5.