人B7-1和B7-2cDNA的克隆及鉴定  

CLONlNG OF B7-1 AND B7-2 GENES FROM HUMAN BURKlTT'S B LYMPHOCYTE LINE

作　　者：张惠丽[1] 奚永志[1] 孔繁华[1] 陈汝光 屠敏[1] 刘楠[1] 

机构地区：[1]军事医学科学院附属医院免疫学研究室,北京100039 [2]深圳市宝安中心血站,广东深圳518101

出　　处：《中国应用生理学杂志》2001年第2期117-120,共4页Chinese Journal of Applied Physiology

基　　金：军队九五医药卫生科研基金及国家自然科学基金!(3990 0 187)资助课题

摘　　要：目的 :为探索性构建全新型重组人B7 PE40绿脓杆菌外毒素融合蛋白以长期诱导免疫耐受 ,本研究从急性B淋巴细胞白血病细胞株Raji中克隆N 末端分别缺失 34和 1 6个氨基酸的人B7 l和B7 2基因胞外区 ,并构建含此基因的重组质粒。方法 :根据B7 1和B7 2基因序列设计合成可扩增B7 1和B7 2cDNA的特异性引物 ,用RT PCR的方法从Raji细胞总RNA中扩增B7 1和B7 2cDNA ,并克隆至 pGEM T载体中 ,经酶切鉴定后再进行序列分析。结果和结论 :从Raji细胞中扩增出预期的 6 2 4和 6 75bp的B7 1和B7 2cDNA ,将其克隆至 pGEM T载体中 ,分别经EcoRI/HindⅢ和BamHI/Sphl双酶切电泳和序列分析确证 ,为进一步构建人B7Aim: In order to construct recombinant B7 Pseudomonas exotoxin fusion protein to induce long term immunotolerance, we tried to clone both B7 1 and B7 2 genes from human Burkitt's B lymphocyte line(Raji) and construct the recombinant plasmids encoding for B7 1 and B7 2. Methods: Two pairs of primers for B7 1 and B7 2 were designed and synthesized according to the sequences of human B7 1 and B7 2 genes derived from genbank. Two recombinant plasmids pGEM T B7 1land pGEM T B7 2 were constructed by recombinant gene techniques. ResuIts and Conclusion: We have successfully cloned both B7 1 and B7 2 cDNA which, confirmed by DNA sequencing, EcoRI/HindⅢ and BamHI/SphI restriction enzyme digesting,could be used to construct recombinant B7 Pseudomonas extoxin fusion proteins.

关 键 词：B7-1 B7-2 基因 淋巴瘤 CDNA 克隆 急性B淋巴细胞白血病 细胞样 

分 类 号：R733.71[医药卫生—肿瘤] R392.3[医药卫生—临床医学]