摘要
构建了人纤维结合素（ＦＮ）的三功能结构域重组多肽的两个表达质粒，分别编码两个重组多肽：ＣＨ６２（ＦＮ的Ｐｒｏ１２３９～Ｓｅｒ１５１５经Ｍｅｔ和Ａｌａ１６９０～Ｖａ１２０４９相连）和ＣＨ６３（从ＣＨ６２中删除了Ｉｌｅ１８５０～Ｇｌｕ１９７８）．ＣＨ６２在大肠杆菌中的表达效率很低，而ＣＨ６３的表达效率则很高，结果提示ＦＮ分子中的Ａｓｐ１９６１～Ｇｌｕ１９７８序列是影响三结构域多肽在大肠杆菌中表达的关键结构．ＣＨ６３经过溶解和复性后，可通过肝素－琼脂糖亲和层析得到纯品，所得纯品具有结合肝素和结合细胞的功能，且结合细胞的能力比双结构域ＦＮ多肽更强，表明两个结合细胞的功能结构域均有活性．ＣＨ６３的制备为进一步研究具有更强的抑制肿瘤转移作用的基因工程制品奠定了基础．
Two plasmids were constructed and used to express two triple domain recombinant polypeptide of human fibronectin(FN).cDNAs in plasmids code for two polypeptides:CH62(Pro1239-Ser1515 of FN linked with Ala 1690-Val 2049 through Met)and CH63(CH62 without Ile1850-Glu1978)respectively.The expression level of CH62 in E.coli was very low,but that of CH63 was very high.The result suggests that Asp1961-Glu1978 in FN is the key sequence which influences the expression of triple domain polypeptide in E.coli. After dissolved and renatured,CH63 can be purified by heperin agarose affinity chromatography.The purified product is capable of binding both heparin and cells.The cell binding capacity of CH63 is much stronger than that of double domain polypeptide of fibronectin,suggesting that both of the cell binding domains in the recombinant polypeptide are functional.The production of CH63 provides a fundermental basis for further study of recombinant product with better antimetastasis function.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1998年第2期122-127,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金
卫生部科学基金
