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首页> 外文期刊>Biotechnology Letters >Establishment of a Bombyx mori nucleopolyhedrovirus (BmNPV) hyper-sensitive cell line from the silkworm e21 strain
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Establishment of a Bombyx mori nucleopolyhedrovirus (BmNPV) hyper-sensitive cell line from the silkworm e21 strain

机译:家蚕e21毒株家蚕核多角体病毒(BmNPV)超敏细胞系的建立

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摘要

Baeuloviral expression systems, including those of Autographa californica multiple nucleopolyhedrovirus Bombyx mori nucleopolyhedrovirus (BmNPV), are used for recombinant protein production. Four B. mon'-derived (BmN4, Bm5, Bmcl40, and Bme21) cell lines were infected with recombinant BmNPV viruses expressing firefly luciferase or EGFP as reporters under the control of a viral polyhedrin promoter. Bme21 exhibited significantly higher (100-fold) luciferase activity than BmN4 and Bm5. With the EGFP reporter protein, Bme21 cells showed a marked increase in the ratio of EGFP-positive cells, reaching 90 % on day 4 post-infection, while Bm5 and BmN4 cells had a slow increase in the ratio of their EGFP-positive population. The viral titer in a supernatant of Bme21 cell culture increased faster than those of Bm5 and BmN4 cells. This susceptibility indicates that the Bme21 cell line is useful for large-scale protein expression using BmNPV.
机译:Baeuloviral表达系统,包括加利福尼亚州的Autographa californica多核多角体病毒Bombyx mori核多角体病毒(BmNPV),用于重组蛋白生产。在病毒多面体启动子的控制下,用表达萤火虫荧光素酶或EGFP作为报告基因的重组BmNPV病毒感染了四个B. mon'衍生的BmN4,Bm5,Bmcl40和Bme21细胞系。 Bme21展示出比BmN4和Bm5高得多的荧光素酶活性(100倍)。有了EGFP报告蛋白,Bme21细胞的EGFP阳性细胞比例显着增加,在感染后第4天达到90%,而Bm5和BmN4细胞的EGFP阳性细胞比例缓慢增加。 Bme21细胞培养上清液中的病毒滴度增加快于Bm5和BmN4细胞。这种敏感性表明,Bme21细胞系可用于使用BmNPV进行大规模蛋白质表达。

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