表达人 HGF/MBP 融合蛋白的 pMAL-MBP/HGF 重组质粒的构建及鉴定  

IDENTIFICATION OF HUMAN HGF/MBP FUSION PROTEIN EXPRESSED BY HUMAN HGF cDNA INSERTED INTO PLASMID pMAL C2

作　　者：方海林[1] 张立煌[1] 孙永良[1] 姚航平[1] 林舜华[1] 李敏伟[1] 

机构地区：[1]浙江医科大学传染病研究所

出　　处：《中国免疫学杂志》1997年第3期135-135,共1页Chinese Journal of Immunology

基　　金：浙江省卫生厅1995年资助

摘　　要：将人ＨＧＦ完整编码区ｃＤＮＡ片段插入ＭＢＰ表达型ｐＭＡＬ－Ｃ２载体质粒中，构建了表达人ＨＧＦ／ＭＢＰ融合蛋白的ｐＭＡＬ－ＭＢＰ／ＨＧＦ重组质粒。表达的ＨＧＦ／ＭＢＰ融合蛋白经ＳＤＳ－ＰＡＧＥ分析分子量约为１１０ｋＤ，Ｗｅｓｔｅｒｎｂｌｏｔｉｎｇ表明能被兔抗ＭＢＰ抗体和抗人ＨＧＦＭｃＡｂ所识别。结果提示可利用该表达型重组质粒制备重组人ＨＧＦ。Coding fragment of human hepatocyte growth factor(HGF) cDNA (75～2 271)was inserted downstream from the mal.E gene of the pMAL C2 vector which encodes maltose binding protein(MBP),resulting in the expression of human HGF/MBP fusion protein with E.coli TB1 strain as host successfully.The molecular weight of this fusion protein is estimated as 110 kD with 10% SDS PAGE.Both specificities of MBP and human HGF co exist in this fusion protein verified with Western blotting using rabbit anti MBP and McAb anti human HGF.Because factor Xa recognization site remains at the linked part between the MBP and HGF of the fusion protein it is simple to separate and purify the recombinant human HGF with corresponding affinity columns.These results reported here have facilitated the use of recombinant human HGF to further investigate the basic biologic activities and clinical applications of HGF.

关 键 词：肝细胞生长因子 pMAL-C2 质粒 HGF/MBP 融合蛋白 

分 类 号：Q78[生物学—分子生物学] R394.8[医药卫生—医学遗传学]