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作 者:韩梅[1] 靳更林[2] 寿成超[2] 戴寿芝[1]
机构地区:[1]宁夏医学院免疫室,宁夏银川750004 [2]北京大学临床肿瘤学院,北京100034
出 处:《宁夏医学杂志》2001年第4期195-196,共2页Ningxia Medical Journal
摘 要:目的 构建能表达抗胃癌单链抗体免疫毒素的重组质粒并进行表达。方法 应用基因工程方法 ,将抗胃癌的单链抗体基因与绿脓杆菌外毒素基因进行重组 ,得到的重组质粒转化大肠杆菌BL - 2 1,鉴定后的阳性克隆经IPTG诱导表达。结果 重组质粒经酶切鉴定正确 ,IPTG诱导后可看到明显的表达带 ,分子大小与预计值相符。结论 得到了能表达抗胃癌单链抗体免疫毒素的重组质粒。Objectives To construct the recombinant plasmid that expresses anti-carcinoma ScFv immunotoxin.Methods By recombinant technique,the cDNA coding the anti-carcinoma ScFv antibody was recombined with the cDNA coding Pseudomonas areuginosa exotoxin to form the recombinanr plasmid.The E Coli.BL-21,after being transformed with the recombinant plasmid,was induced by IPTG to express the anti-carcinoma ScFv immumotoxin.Results The recombinantion of plasmid was identifide to be correct by endonuclease restriction analysis and the BL-21 that contain the recombinant plasmid abundantly expressed the immunotoxin by IPTG inducement.The size of the expressed protein is similar to that anticipated in MW.Conclusions The recombinant plasmid that can express the anti-carcinoma ScFv immunotoxin is successfully obtained.
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