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杆状病毒介导的细菌基因在双翅目和哺乳动物细胞中的表达。

Baculovirus-mediated expression of bacterial genes in dipteran and mammalian cells.

作者信息

Carbonell L F, Klowden M J, Miller L K

出版信息

J Virol. 1985 Oct;56(1):153-60. doi: 10.1128/JVI.56.1.153-160.1985.

DOI:10.1128/JVI.56.1.153-160.1985
PMID:2993646
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC252500/
Abstract

A recombinant baculovirus containing the Escherichia coli chloramphenicol acetyltransferase (CAT) gene under the control of the Rous sarcoma virus long terminal repeat promoter and the E. coli beta-galactosidase gene under the control of the very late baculoviral polyhedrin promoter was used to determine if Autographa californica nuclear polyhedrosis virus, a baculovirus of Lepidoptera, can enter and express viral DNA in dipteran (Drosophila sp.) and mammalian (Mus sp.) cells that are considered refractory to baculovirus replication. Following infection, CAT gene expression was observed in both dipteran and mammalian cells, but expression in the mammalian cell line was less than 0.05% of that observed in either dipteran or lepidopteran cells. Although the level of CAT gene expression was similar in permissive lepidopteran and nonpermissive dipteran cells, expression of beta-galactosidase activity from the late polyhedrin promoter in dipteran or mammalian cells was less than 0.3% of the levels observed in lepidopteran cells. These results indicate that foreign gene expression in nonpermissive cells is promoter dependent and that late viral gene expression is restricted in these cells. The Rous sarcoma virus long terminal repeat allows substantial CAT gene expression in both a D. melanogaster cell line and Aedes aegypti midgut cells. Baculovirus DNA undergoes a limited number of replications in Drosophila cells. The results are relevant to baculovirus host range, the safety of baculoviruses as pesticides, and the development of baculovirus pesticides with expanded host ranges.

摘要

一种重组杆状病毒被用于确定鳞翅目的苜蓿银纹夜蛾核型多角体病毒是否能在被认为对杆状病毒复制具有抗性的双翅目(果蝇属)和哺乳动物(小家鼠属)细胞中进入并表达病毒DNA。该重组杆状病毒含有受劳氏肉瘤病毒长末端重复启动子控制的大肠杆菌氯霉素乙酰转移酶(CAT)基因,以及受杆状病毒极晚期多角体蛋白启动子控制的大肠杆菌β-半乳糖苷酶基因。感染后,在双翅目和哺乳动物细胞中均观察到了CAT基因表达,但在哺乳动物细胞系中的表达量不到在双翅目或鳞翅目细胞中观察到的表达量的0.05%。尽管在允许性的鳞翅目细胞和非允许性的双翅目细胞中CAT基因表达水平相似,但双翅目或哺乳动物细胞中来自晚期多角体蛋白启动子的β-半乳糖苷酶活性表达量不到在鳞翅目细胞中观察到的水平的0.3%。这些结果表明,非允许性细胞中的外源基因表达是启动子依赖性的,并且这些细胞中晚期病毒基因表达受到限制。劳氏肉瘤病毒长末端重复序列允许在黑腹果蝇细胞系和埃及伊蚊中肠细胞中大量表达CAT基因。杆状病毒DNA在果蝇细胞中进行有限次数的复制。这些结果与杆状病毒的宿主范围、杆状病毒作为杀虫剂的安全性以及扩大宿主范围的杆状病毒杀虫剂的开发有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f343/252500/52bcb4041b0a/jvirol00115-0167-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f343/252500/52bcb4041b0a/jvirol00115-0167-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f343/252500/52bcb4041b0a/jvirol00115-0167-a.jpg

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