Effect of fragment Asp1961—Glu1978 in fibronectin on the expression of triple-domain polypeptide inE. coli

Abstract

Two plasmids were constructed and used to express two triple-domain recombinant polypeptide of human fibronectin (FN). The cDNAs in plasmids code for two polypeptides, CH62 (Pro1239-Ser1515 of FN linked with Ala1690-Val2049 through Met) and CH63 (CH62 without Ile1850-Glu1978). The expression level of CH62 inE. coli was very low, but that of CH63 was very high. The results suggests that Asp1961 -Glu1978 in FN is a key sequence influencing the expression of triple-domain polypeptide inE. Coli. After being dissolved and renatured, CH63 can be purified by heparin-agarose affinity chromatography. Both of the cell-binding domains in the recombinant polypeptide were functional. The production of CH63 provides a fundamental basis for further study of recombinant products with better anti-metastasis function.

Keywords

fibronectin / recombinant polypeptide / E. coli / expression