作　　者： ; ; ; ;

机构地区： 广州医学院第二附属医院

出　　处： 《山东大学耳鼻喉眼学报》 2006年第1期1-3,8,共4页

摘　　要： 目的:克隆人B7-1全长cDNA以构建相应腺病毒表达载体。方法:根据B7-1基因序列设计并合成可扩增B7-1 cDNA的特异性引物,用RT-PCR法从骨髓细胞总RNA中扩增B7-1 cDNA,并克隆至pGEM-T载体中,经酶切鉴定后再行序列分析。结果:逆转录多聚酶链反应扩增产物长度与预期的889 bp一致;用M13正、反向引物行序列测定,证实克隆出的序列与GenBank的B7-1 cDNA序列完全一致;人B7-1全长cDNA被成功地插入到质粒pGEM-T中。结论:克隆人B7-1全长cDNA为构建相应腺病毒表达载体及应用B7-1行肿瘤免疫基因治疗提供了可能性。 Objective： To construct recombinant B7-1 retrovirus expressing vector by clone of B7-1 gene from human blood and construct the recombinant plasmid of B7-1, Method： Primers for B7-1 were designed and synthesized according to the sequence of human B7-1 gene derived from GenBank, The full length cDNA of B7-1 was cloned by RT-PCR techniques, The recombinant plasmid pGEM-T- B7 were constructed by DNA recombinant techniques. Results： The length of RT-PCR product coincided with that of authors＇ anticipation（889 bp）, and the recombinant plasmid was confirmed by restriction enzyme digesting with EcoRI and Hind Ⅲ. 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 pGEM-T vector. Conclusion： The successful cloning of human B7-1 cDNA, as well as the construction of its retrovirus expressing vector enables us to further investigate the role of B7-1 in tumor immunogene therapy.

关 键 词： 受体 基因 克隆

领　　域： [生物学]