전체메뉴
검색
Article Search

JMB Journal of Microbiolog and Biotechnology

QR Code QR Code

Related articles in JMB

More Related Articles

Article

J. Microbiol. Biotechnol. 2002; 12(1): 65-70

Published online February 28, 2002

Copyright © The Korean Society for Microbiology and Biotechnology.

Expression and Receptor Binding Activity of Fusion Protein from Transforming Growth Factor-β1 and GFP

Jun Ho Yoon , Pyeung Hyeun Kim , Gie Taek Chun , Eui Yul Choi 1 and Se Won Yie 1*

Division of Life Sciences College of Natural Sciences Kang Won National University Chun Cheon Kang Won Do 200-701 Korea, 1Department of Genetic Engineering College of Natural Sciences Hallym University Chun Cheon Kang Won Do 200-702 Korea

Abstract

A TGF-${\beta}1$/GFP monomeric fusion protein was cloned from pPK9A and pGFP-Cl plasmid by PCR amplification. The fusion protein was expressed in a $Bac-To-Bac^{TM}$ baculovirus expression system. A 45 kDa fusion protein was purified using an Ni-NTA column with 300 mM imidazol from a cell lysate infected with recombinant viruses for 72 h post-infection. The fusion protein cross-reacted with the commercial $TGF-{\beta}1$ polyclonal Ab as well as Ab raised against a precursor, monomeric $TGF-{\beta}1$, and GFP. The binding activity of the fusion protein with a $TGF-{\beta}1$ receptor was examined. Fluorescence was observed in Mv1Lu cells, yet not in insect cells treated with the fusion protein. No fluorescence was detected in Mv1Lu cells incubated with the fusion protein treated with Ab prior to the binding reaction, or with GFP alone, thereby indicating that the binding of the fusion protein was specific to $TGF-{\beta}1$ with a receptor.

Keywords: TGF-b1/GFPfusionprotein|baculovirusexpression|receptorbinding