RRC ID 29180
Author Tsuchiya T, Tojo A, Shibuya M.
Title Multi-copy introduction and high-level expression of interleukin-3 genes by retroviral vector superinfection.
Journal Biochem Biophys Res Commun
Abstract We constructed a retroviral expression vector carrying multiple cloning sites. This vector was found to express efficiently the cloned gene. Using this vector and a helper virus-free system, a murine interleukin-3 (mlL-3) high-producing cell line was established by multiple cycles of infection with recombinant retroviruses carrying mlL-3 cDNA. The infected cells produced a considerable amount of mlL-3 and the concentration of mlL-3 in culture media increased as a function of the frequency of infection. High levels of mlL-3 cDNA, mRNA and protein in this cell line were confirmed by Southern, Northern and biological assays, respectively. These results suggest that artificial gene amplification is possible in a helper-free retroviral system. This should be applicable to efficient expression of bioactive molecules in a wide variety of mammalian cells including suspension cells.
Volume 158(2)
Pages 576-83
Published 1989-1-31
DOI 10.1016/s0006-291x(89)80088-9
PII S0006-291X(89)80088-9
PMID 2783848
MeSH Animals Blotting, Northern Blotting, Southern Genetic Engineering Genetic Vectors* Interleukin-3 / genetics* Mice Recombinant Proteins Retroviridae / genetics Superinfection
IF 2.985
Times Cited 4
DNA material pMNSM (RDB03043)