Use of helper-free retroviral vector to direct a high expression of porcine growth hormone in mouse fibroblast cells.
A retroviral vector has been employed to express the cDNA coding for porcine growth hormone (pGH) in the mouse fibroblast cell NIH 3T3 in large quantity. In this study, a single gene vector which contained no selectable marker was used. We have coinfected NIH 3T3 cells with pGH retrovirus and Neo(r) retrovirus to obtain a stable, high-expression clone. Using a superinfection strategy, we further increased the copy number of proviral DNA in the host chromosome, thus increasing the pGH secretion from 22 to 55 micrograms/10(6) cells/24 h. The recombinant pGH produced from mouse fibroblast cells was heterogeneous at the N-terminus, which mimicked the situation with bovine growth hormone either from natural sources or from recombinant products derived from mouse fibroblasts. This technology is useful for many biologically important genes to be stably transduced by retroviral vector into mammalian cells and highly expressed.
Hwang
L H
LH
Molecular Biology Division, Development Center for Biotechnology, Taipei, Taiwan, Republic of China.
Chen
B F
BF
Lee
P J
PJ
Ho
S Y
SY
Liu
J J
JJ
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001846:Bone Development / Q000187:drug effects; D003001:Cloning, Molecular; D004247:DNA / Q000096:biosynthesis; D005822:Genetic Vectors; D013006:Growth Hormone / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013552:Swine
D011994:Recombinant Proteins; D013006:Growth Hormone; D004247:DNA