人B7.1基因cDNA的克隆及其逆转录病毒表达载体的构建

The Clone of Human B7.1 cDNA and Its Construction with Retrovirus Vector

目的 克隆人 B7.1全长 c DNA及构建相应的逆转录病毒表达载体。 方法 应用逆转录多聚酶链反应从人 B淋巴瘤细胞系 Raji中克隆 B7.1全长 c DNA ,再将 c DNA插入到质粒 p Bluescript中 ,经自动荧光测序证实无误后 ,再通过定向克隆构建相应的逆转录病毒表达载体 PL XSNh B7。 结果 逆转录多聚酶链反应扩增产物长度与预期的 889bp一致 ;用 M13正、反向引物进行荧光测序证实 ,克隆出的序列与 Gen Bank的 B7.1c DNA序列完全一致 ;人 B7.1全长 c DNA被成功地插入到质粒 PL XSN中。 结论 人 B7.1全长 c DNA的克隆及相应逆转录病毒表达载体的构建为今后应用 B7.1进行肿瘤免疫基因治疗提供了可能性。

Objective\ To clone full length cDNA of human B7.1 and construct its retrovirus expressing vector.\ Methods\ The full length cDNA of B7 1 was cloned by RT\|PCR from human B lymphoma Raji cell line and confirmed by DNA sequencing.\ Then,the recombinant retrovirus expressing vector was constructed by inserting the cDNA into PLXSN.\ Results\ The length of RT\|PCR product coincided with that of our anticipation(889 bp), and the sequencing result of the cDNA was identical to the sequence of B7 1 cDNA in GenBank, and the full length cDNA of human B7 1 was successfully inserted \{into\} the vector of PLXSN.\ Conclusion\ The successful cloning of human B7 1 cDNA as well as construction of its retrovirus expressing vector enables us to further investigate the role of B7 1 in tumor immunogene therapy.\;