Volume 11, No 2, Jun 2001
ISSN: 1001-0602
EISSN: 1748-7838 2018
impact factor 17.848*
(Clarivate Analytics, 2019)
Volume 11 Issue 2, June 2001: 95-100
ORIGINAL ARTICLES
The expression and antigenicity identification of recombinant rat TGF-β1 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
Correspondence:
In order to study structure-function details of TGF-β1,the recombinant mature form of rat TGF-β 1 was expressed in bacteria. Synthesis of the 112 amino-acid carboxyl-terminal part of TGF-β1 (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-β1. The molecular weight of expressed TGF-β1 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-β1 expressed in this study provides a useful tool for future detailed structural and functional studies.
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