肿瘤防治研究.pdfVIP

肿瘤防治研究 2007 年第 34 卷第 6 期 4·09 · 凋亡抑制蛋白L ivin 两种异构体原核表达载体 的构建及融合表达 邹爱民 ,沈建军 ,高 　萍 ,林 　芳 ,张惠中 Construction of Prokaryotic Expression Vectors f or L ivin Alpha and L ivin Beta ZOU Aimin ,SH EN J ianj un , GAO Ping ,L IN Fang , ZHAN G Huizhong Cent ral L aboratory , Tang d u H osp i t al , Fou rt h M i l i t a ry M e d ical Uni ve rs i ty , X i′an 7 10038 , Chi na Corresp on d i ng A ut hor : Z HA N G H uiz hong , Em ai l : hongy u200 1 @163 . com α β Abstract :Objective 　To con st ruct p rokaryotic exp ression vector s for Livin and Livin and to obt ained t he ( ) α ( ) β fu sion p rot ein of p E T32a + livin and p E T32a + livin . Methods 　Tot al RN A of Hela cell wa s ex t ract ed . The fulllengt h cDN A of Livin i soforms wa s gained by R TPCR . Then in sert ed into p E T32a ( + ) ( ) α α ( ) β vector and con st ruct ed t he recombinant p la smids p E T32a + livin / D H5 and p E T32a + / livin / α α D H5 , sequencing wa s p erformed to guarant ee correct sequence in sertion for t he i soform s Livin and β ( ) α ( ) β Livin . Recon st ruct ed t he recombinant p la smids p E T32a + livin / BL 2 1 and p E T32a + / livin / ( ) α ( ) β BL 2 1 ,t he p E T32a + livin / BL 2 1 and p E T32a + livin / BL 2 1 p la smids wa s induced aft er 4 . 5 hour s ( ) α by IP T G , and t he value of A600nm wa s 0 . 6 in LB medium . Exp ression of t he p E T3