| Title | Expression of immunologically active porcine recombinant TGF-β1 precursor protein in baculovirus system |
|---|---|
| Author | Lim, Hyun · Kim, Pyeung Hyeun · Chun, Gie Taek · Choi, Eui Yul¹ · Yie, Se Won * |
| Address | Department of Microbiology, College of Natural Sciences, Kangwon National University; ¹Department of Genetic Engineering, College of Natural Sciences, Hallym University |
| Bibliography | Journal of Microbiology, 35(4),341-346, 1997, |
| DOI | |
| Key Words | TGF-β1, baculovirus expression, recombinant protein, precursor |
| Abstract | In order to express recombinant porcine TGF-β1 protein in a baculovirus expression system the entire TGF-β1 gene containing extra amino acids at the N terminus was cloned into pFBa and pFBb of the Bac-To-Bac^TM baculovirus expression system. One of the clones contained 106 extra amino acids and was designated pFBa-106 TGF-β1, and the other had 28 extra amino acids and was designated pFBb-28 TGF-β1. The orientation of the gene was identified with restriction enzyme mapping and PCR with internal TGF-β1 primers. Sf-9 cells were infected at a m.o.i. of 10 by the recombinant viruses generated from the two expected sizes of 55 kD and 46.4kD. these precursor forms of TGF-β1 with a polyclonal antibody against human TGF-β1. No mature form of TGF-β1 protein was detected on SDS gels and an immunoblot indicated that TGF-β1 precursor is not properlu processed in insect cells. |
| Download PDF | Eng_350416_341-346p.pdf |