Premium
bcl‐2 expression in Spodoptera Frugiperda Sf‐9 and Trichoplusia Ni BTI‐Tn‐5B1‐4 insect cells: Effect on recombinant protein expression and cell viability
Author(s) -
MitchellLogean Christine,
Murhammer David W.
Publication year - 1997
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19971120)56:4<380::aid-bit4>3.0.co;2-k
Subject(s) - autographa californica , trichoplusia , coinfection , biology , microbiology and biotechnology , recombinant dna , spodoptera , virology , virus , noctuidae , gene , biochemistry , botany , larva
The effect of bcl‐2 expression on cell viability and recombinant protein synthesis was investigated in the Spodoptera frugiperda Sf‐9 and Trichoplusia ni BTI‐Tn‐5B1‐4 (High Five™) insect cell lines. It was found that coinfection with a baculovirus expressing bcl‐2 [ Autographa californica nuclear polyhedrosis virus (AcNPV)‐bcl2] extended the life span of High Five™ cells but not Sf‐9 cells when compared to infection with recombinant baculoviruses expressing either human tissue plasminogen activator (AcNPV‐tPA) or Escherichia coli β‐galactosidase (AcNPV‐βgal). Similar results were obtained in coinfection experiments; i.e., AcNPV‐bcl2 coinfection increased the life span of High Five™ cells over that of cells infected with either AcNPV‐tPA or AcNPV‐βgal alone, but they did not affect the life span of coinfected Sf‐9 cells. Coinfection of Sf‐9 cells with AcNPV‐bcl2 and AcNPV‐βgal resulted in a decrease in the maximum β‐gal expression levels of over 90% when compared to infection with AcNPV‐βgal alone. A similar trend was found in the β‐gal mRNA levels. Coinfection also resulted in a reduced β‐gal expression level in High Five™ cells, but the reduction was consistent with what would be expected when two recombinant viruses compete for use of the cellular machinery. In contrast to the inhibitory effect of AcNPV‐bcl2 coinfection on βgal expression, t‐PA expression levels were either not affected (Sf‐9 cells) or were increased 50% (High Five™ cells) over those obtained by infection with AcNPV‐tPA alone. These results support the hypotheses that bcl‐2 can inhibit transcription of genes under polyhedrin promoter control and that β‐gal expression levels, but not t‐PA expression levels, are controlled at the transcriptional level. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 56: 380–390, 1997.