| 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
|
|
WOS Category
|
BIOPHYSICS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| DNA material |
pMNSM (RDB03043) |
