|
 |
Cell Research (2001); 11(2):95-100  |
The expression and antigenicity identification of recombinant rat TGF-b1 in bacteria
GAO Chun Fang1, Xian Tao KONG1, Axel M GRESSNER2, Ralf WEISKIRCHEN2,*
1Department of Laboratory Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
2Institute of Clinical Chemistry and Pathobiochemistry, Central Laboratory, RWTH-University Hospital, Pauwelsstr 30, D-52074 Aachen, Germany
ABSTRACT
In order to study structure-function details of TGF-b1,the recombinant mature form of rat TGF-b 1 was expressed in bacteria. Synthesis of the 112 amino-acid carboxyl-terminal part of TGF-b1 (amino acid 279-390) was controlled by an inducible gene expression system based on bacteriophage T7 RNA polymerase. This system allowed an active and selective synthesis of recombinant TGF-b1. The molecular weight of expressed TGF-b1 monomer determined on SDS-polyacrylamide gel under reducing conditions was about 13 kD. Serial detergent washes combined with a single gel-filtration purification step were sufficient to purify the expression product to homogeneity. Amino-terminal sequencing revealed that the N-terminal of the recombinant protein was identical to the published data. In Western blot analysis the recombinant polypeptide showed excellent antigenicity against polyclonal TGF-b1 antibody. The mature recombinant rat TGF-b1 expressed in this study provides a useful tool for future detailed structural and functional studies.
Key words: TGF-b1, recombinant expression, inclusion bodies, purification, antigenicity.
Table of Contents
Download as printable (PDF) file
Full Text (html)file
copyright©2006 2006 Institute of Biochemistry and Cell Biology,SIBS,CAS
ISSN:1001-0602(Print),1748-7838(Online);CN:31-1568
suggested resolution 1024*76