대장균내에서 발현된 돼지 TGF-$\beta$1의 분리 및 면역학적 항원성 보유검증

  • 최은영 (한림대학교 의과대학 생화학교실) ;
  • 김현태 (삼진제약주식회사 중앙연구소) ;
  • 김평현 (강원대학교 자연과학대학 생명과학부) ;
  • 변우현 (강원대학교 자연과학대학 생명과학부)
  • 발행 : 1997.04.01

초록

Porcine transforming growth factor-$\beta$1 (TGF-$\beta$1) was expressed in Escherichia coli using cDNA of TGF-$\beta$1 and glutathione S-transferase (GST) fusion vector pGEX-1$\lambda$T. An ApoI-Tth111I fragment of cDNA which correspond to the amino acid residues from 123 to 390 of the precursor TGF-$\beta$1 was inserted into EcoRI-Tth111I digested pGEM#-l$\lambda$T and the recombined plasmid was named pGET-12. Gene products from the cloned regions of the recombinant plasmids pGET-12 was not detected in soluble fraction of cell free extract but detected in insoluble fraction. The solubilization of insoluble gene product was achieved by the treatment of N-laurylsarcosine. Molecular weight of partially purified proteins determined by electrophoresis was same as expected from cloned fragment. The ELISA test results of the purified proteins showed that immunologically detectable epitope was preserved in recombinant protein.

키워드

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