FOREIGN GENE-EXPRESSION BY A BACULOVIRUS VECTOR WITH AN EXPANDED HOST RANGE
FOREIGN GENE-EXPRESSION BY A BACULOVIRUS VECTOR WITH AN EXPANDED HOST RANGE
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DOI：
10.1099/0022-1317-73-7-1877
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发表时间：
1992-07-01
影响因子：
3.8
通讯作者：
MATSUBARA, F
中科院分区：
文献类型：
--
作者：
MORI, H;NAKAZAWA, H;MATSUBARA, F
利用DNA重组技术改进了基于核型多角体病毒（NPV）的基因表达系统。家蚕BmN细胞系和草地贪夜蛾Sf 21细胞系对苜蓿银纹夜蛾多衣壳核型多角体病毒（AcMNPV）和B均不敏感。家蚕核型多角体病毒（BmNPV）复制。将AcMNPV DNA和BamHI消化的BmNPV DNA共转染Sf 21细胞后，通过BmN细胞单层上的噬斑纯化分离子代病毒，并分析一种病毒分离物的宿主特异性。该病毒具有较广的宿主范围，能在Sf 21细胞、BmN细胞和家蚕B幼虫中复制和产生多角体。森。DNA限制性内切酶分析表明，该分离物为AcMNPV和BmNPV的杂交种。使用AcMNPV转移载体pAcYM 1，将杂合病毒的多角体蛋白基因的一部分用萤火虫荧光素酶基因的编码区替换，产生重组病毒。后者在多角体蛋白启动子的控制下在两种细胞系和家蚕幼虫中表达萤火虫荧光素酶。
A nuclear polyhedrosis virus (NPV) (Baculoviridae)-based gene expression system was improved by DNA recombination. The BmN cell line established from Bombyx mori and the Sf21 cell line established from Spodoptera frugiperda are non-permissive for Autographa californica multicapsid NPV (AcMNPV) and B. mori NPV (BmNPV) replication, respectively. After cotransfection of AcMNPV DNA and BamHI-digested BmNPV DNA into Sf21 cells, progeny viruses were isolated by plaque purification on BmN cell monolayers and the host specificity of one viral isolate was analysed. The virus had a wider host range, and replicated and produced polyhedra in Sf21 cells, BmN cells and larvae of the silkworm, B. mori. DNA restriction endonuclease analysis showed that the isolate was a hybrid of AcMNPV and BmNPV. Using the AcMNPV transfer vector pAcYM 1 a portion of the polyhedrin gene of the hybrid virus was replaced with the coding region of the firefly luciferase gene, producing a recombinant virus. The latter expressed firefly luciferase in both cell lines and in silkworm larvae under the control of the polyhedrin promoter.