INSECT CELL HOSTS FOR BACULOVIRUS EXPRESSION VECTORS CONTAIN ENDOGENOUS EXOGLYCOSIDASE ACTIVITY

被引:52
作者
LICARI, PJ
JARVIS, DL
BAILEY, JE
机构
[1] CALTECH,DEPT CHEM ENGN,PASADENA,CA 91125
[2] TEXAS A&M UNIV SYST,DEPT ENTOMOL,COLL STN,TX 77843
关键词
D O I
10.1021/bp00020a005
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Four different insect cell lines that can be used as hosts for baculovirus infection were assayed for the presence of endogenous exoglycosidases. All four cell lines, derived from Spodoptera frugiperda, Trichoplusia ni, Bombyx mori, or Malacosoma disstria, contained N-acetyl-beta-glucosaminidase, N-acetyl-beta-galactosaminidase, beta-galactosidase, and sialidase activities. Exoglycosidase activities were found in cell lysates as well as cell-free supernatants from uninfected and wild-type baculovirus infected cells. Oligosaccharide analysis of cellular glycoproteins using lectins recognizing Galbeta1, 3GalNAc, Galbeta1,4GlcNAc, and NeuAcalpha2,6Gal demonstrated that only Galbeta1,3GalNAc was present. The demonstration that these cells contain exoglycosidases raises the possibility that the oligosaccharides of baculovirus-expressed glycoproteins are subject to enzymatic degradation.
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页码:146 / 152
页数:7
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