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Cloning and Expression of SpltMNPV

Cloning and Expression
of SpltMNPV
Sl136 Gene and Functions
of the Expressed Product

ZHANG Ping, YANG Bo, PANG Yi*, SU De-Ming1
(State Key Laboratory for Biocontrol, Zhongshan University, Guangzhou
510275, China
;1Virology Research Unit, Fudan University,
Shanghai
200433,
China
)

Abstract    By
computer-assisted analysis, it was revealed that ORF136 gene product in
SpltMNPV genome had the basic properties of membrane protein. A putative signal
peptide was present at the N-terminal and a transmembrane region near the
C-terminal of SL136 protein. In the N-terminal half region, there was a
coiled-coil domain, which is a typical feature of a number of viral fusion
proteins. After PCR amplification, a recombinant plasmid pBVSl136 and
a recombinant AcMNPV containing Sl136 were constructed, in order to
express Sl136 gene in E.coli and insect Hi5 cells,
respectively. The SDS-PAGE results showed that both expression levels were
high. Cell membrane fusion was induced in the Sl-zsu-1 cells,whichhad been
transfected with Sl136 gene alone, by lowering pH of the medium. These
results suggested that SL136 protein may be an envelope fusion protein.
Key words    Spodoptera litura
; baculovirus; fusion protein; expression

*Corresponding author: Tel, 86-20-84113860; Fax, 86-20-84037472; e-mail, [email protected]