肿瘤坏死因子相关凋亡诱导配体基因原核表达载体的构建、表达与抗体制备  

Construction,expression and preparation of antibody of TRAIL

作　　者：张旭[1] 孙等军[2] 方琴[3] 任志娟[1] 陈剑[1] 肖芸[1] 王季石[1] 

机构地区：[1]贵阳医学院附属医院血液科,550004 [2]贵阳医学院附院肿瘤科 [3]贵阳医学院附院药剂科

出　　处：《贵州医药》2003年第7期579-582,共4页Guizhou Medical Journal

基　　金：省长专项基金(No.S2001-14)

摘　　要：目的 获取抗肿瘤坏死因子相关凋亡诱导配体(TRARAIL)抗体。方法 利用PCR技术和基因重组技术将TRAIL基因构建于原核表达载体pPre-TMHTb中,经酶切、测序鉴定证实构建完全正确后,通过IPTG诱导其在大肠杆菌DH-5α中获得表达,并将TRAIL蛋白免疫家兔,通过DotBlot及 Western Blot检测抗 TRAIL抗体的产生。结果 成功构建TRAIL基因原核表达载体,并在大肠杆菌DH-5α中获得表达,TRAIL蛋白表达量占菌体蛋白质总量的11.8％,经Dot Blot及 WesternBlot捡测证实获得了抗TRAIL抗体。结论 成功制备抗TRAIL抗体,从而为TRAIL在真核细胞水平的深入研究奠定了实验基础。Objective To obtain the anti-TRAIL antibody. Methods A prokaryotic expression vector pPre-TMHTb was constructed by PCR and recombinant DNA techniques, then it was tested by digestive identification and measuring sequence. TRAIL protein was expressed in E. coli DH-5α through IPTG induced. The rabbits were immunized with TRAIL protein, anti-TRAIL antibody was tested by Dot Blot and Western Blot. Results A prokaryotic expression vector pPre-TMHTb was constructed and expressed in E. coli DH-5α. TRAIL protein was detected up to 11. 8% of the total bacterial protein expressed and anti-TRAIL antibody was tested by Dot Blot and Western Blot. Conclusion The anti-TRAIL antibody was obtained that provide an experimental basis for the studies of TRAIL in eukary-otic level.

关 键 词：肿瘤坏死因子相关凋亡诱导配体 基因表达 大肠杆菌 聚合酶链反应 基因重组技术 

分 类 号：R394[医药卫生—医学遗传学]