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BmNPV chitinase gene deletion enhances foreign gene expression in a BmN cell system
Author(s) -
WANG Fang,
ZHANG ChuanXi
Publication year - 2009
Publication title -
entomological research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.421
H-Index - 20
eISSN - 1748-5967
pISSN - 1738-2297
DOI - 10.1111/j.1748-5967.2009.00192.x
Subject(s) - polyhedrin , chitinase , biology , microbiology and biotechnology , gene , gene expression , heterologous expression , polyacrylamide gel electrophoresis , recombinant dna , genetics , enzyme , biochemistry , spodoptera
Bombyx mori nucleopolyhedrovirus (BmNPV) is extensively being studied as an expression vector for heterologous gene expression in silkworm‐derived cells as well as in the host larvae or pupae. BmNPV chitinase is necessary for liquefaction of the virus‐infected host insect. The influence of chitinase on the efficiency of foreign gene expression was studied to provide a scientific basis for improving the BmNPV expression system. The BmNPV chitinase gene ( chiA ) was deleted and the expression level of the polyhedrin promoter controlling the lac Z gene in BmN cells was determined. Sodium dodecylsulfate (SDS)–polyacrylamide gel electrophoresis (PAGE) showed that β‐galactosidase accounted for approximately 6.9 and 7.7% of the total protein in BmN cells infected with the chiA deficient Bm lac Z + chiA − at 3 and 4 days post infection, while the total protein was 3.2 and 4.2% in cells infected with Bm lac Z + . The relative β‐galactosidase activities in Bm lac Z + chiA − ‐infected cells improved 2.33‐ and 1.56‐fold compared to those of Bm lac Z + ‐infected cells at 3 and 4 days post infection. The results of the present study suggest that chitinase deletion could improve the lacZ expression level in the BmNPV‐BmN cell expression system.