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对缺乏功能性cg30基因的重组杆状病毒的体内和体外分析。

In vivo and in vitro analyses of recombinant baculoviruses lacking a functional cg30 gene.

作者信息

Passarelli A L, Miller L K

机构信息

Department of Genetics, University of Georgia, Athens 30602.

出版信息

J Virol. 1994 Feb;68(2):1186-90. doi: 10.1128/JVI.68.2.1186-1190.1994.

Abstract

The cg30 gene of Autographa californica nuclear polyhedrosis virus (AcMNPV) encodes two sequence motifs, a zinc finger-like motif and a leucine zipper, found in other polypeptides known to be involved in gene regulation. To gain insight into the function of the cg30 product, CG30, we constructed and characterized recombinant viruses lacking a functional cg30 gene. We found that cg30 mutants had no striking phenotype in cell lines derived from Spodoptera frugiperda or Trichoplusia ni or in T. ni larvae. Although cg30 is known to be transcribed as an early monocistronic RNA and as the second cistron of an abundant late bicistronic RNA, production of a CG30-beta-galactosidase fusion protein was observed mainly at early times postinfection. Viruses containing cg30 had a subtle growth advantage over those lacking cg30 after several viral passages in cell culture. We employed transient expression assays to determine whether cg30 and pe-38, an AcMNPV gene that encodes a polypeptide with zinc finger-like and leucine zipper motifs similar to those of cg30, have redundant functions. Although pe-38 may have a role in AcMNPV gene expression, there was no indication that cg30 and pe-38 are functionally redundant.

摘要

苜蓿银纹夜蛾核型多角体病毒(AcMNPV)的cg30基因编码两个序列基序,即锌指样基序和亮氨酸拉链,在已知参与基因调控的其他多肽中也有发现。为了深入了解cg30产物CG30的功能,我们构建并鉴定了缺乏功能性cg30基因的重组病毒。我们发现,cg30突变体在源自草地贪夜蛾或粉纹夜蛾的细胞系以及粉纹夜蛾幼虫中没有明显的表型。尽管已知cg30转录为早期单顺反子RNA以及丰富的晚期双顺反子RNA的第二个顺反子,但主要在感染后早期观察到了CG30-β-半乳糖苷酶融合蛋白的产生。在细胞培养中经过几次病毒传代后,含有cg30的病毒比缺乏cg30的病毒具有轻微的生长优势。我们采用瞬时表达试验来确定cg30和pe-38(AcMNPV的一个基因,其编码的多肽具有与cg30类似的锌指样和亮氨酸拉链基序)是否具有冗余功能。尽管pe-38可能在AcMNPV基因表达中起作用,但没有迹象表明cg30和pe-38在功能上是冗余的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b8fc/236558/fe37bbc00b8e/jvirol00011-0619-a.jpg

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