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KMID : 1041219970390020180
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Journal of Sericultural and Entomological Science
1997 Volume.39 No. 2 p.180 ~ p.185
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Construction of the Novel Baculovirus Transfer Vector Using the p10 Gene of BmNPV
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강석우/Kang, Seok Woo
진병래/윤은영/김상현/김근영/강석권/Jin, Byung Rae/Yun, Eun Young/Kim, Sang Hyun/Kim, Keun Young/Kang, Seok Kwon
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Abstract
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To develope the novel baculovirus transfer vector, the p10 gene was cloned from the Bombyx mori nuclear polyhedrosis virus (BmNPV) vB2 strain isolated from the B. mori larvae of sericultural farms. The novel transfer vector was constructed by using the p10 gene of BmNPV vB2 strain and its function was tested. Open reading frame of the p10 gene from BmNPV vB2 strain was 210 bp. The TAAG sequence at the -71 bp of upstream from translation initiator ATG and two polyadenylation signal site at the downstream from terminator TAA were also detected in the p10 gene. The 5´and 3´ flanking region of the p10 gene amplified by PCR was cloned into pBluescriptII SK(+) and then transfer vector pBm10 was construceted. The 7.9 kb pBm10 was analysed by restriction enzymes and the map was confirmed. In order to determine the expression of foreign gene of pBm10, ß-galactosidase gene was inserted in the Small site of foreign gene cloning site of pBm10. The pBm10 containing ß-galactosidase gene was cotransfected with genomic DNA of BmNPV vB2 into BmN-4 cells. The recombinant baculovirus expressing ß-galactosidase was also produced polyhedra in the infected cells. The results indicated that pBm10 is functional, suggesting that in the baculovirus expression vector system, the recombinant virus produced by pBm10 was effective by oral infection for the producing recombinant proteins in in vivo expression.
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