Effect of fragment Asp1961-Glu1978 in fibronectin on the expression of triple-domain polypeptide in E. coli.
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 in E. 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 in E. 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.
Li
D
D
Department of Medical Molecular Biology, Tongji Medical University, Wuhan.
Feng
Z
Z
Zhang
G
G
Zhang
H
H
Fan
Q
Q
D000818:Animals; D002448:Cell Adhesion; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005353:Fibronectins / Q000096:biosynthesis*; D005818:Genetic Engineering; D006801:Humans; D008546:Melanoma, Experimental / Q000473:pathology; D051379:Mice; D010452:Peptide Biosynthesis; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis
D005353:Fibronectins; D011994:Recombinant Proteins