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J Biotechnol. 2016 Aug 10;231:72-80. doi: 10.1016/j.jbiotec.2016.05.037. Epub 2016 May 28.

Screening and optimization of an efficient Bombyx mori nucleopolyhedrovirus inducible promoter.

Author information

1
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400716, China.
2
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400716, China; Institute of Life Sciences, Chongqing Medical University, Chongqing 400716, China.
3
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400716, China; Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University, Chongqing 400716, China. Electronic address: lucheng@swu.edu.cn.
4
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400716, China; Key Laboratory for Sericulture Functional Genomics and Biotechnology of Agricultural Ministry, Southwest University, Chongqing 400716, China. Electronic address: pmh047@126.com.

Abstract

Pathogen-inducible promoters have been studied extensively and widely used in resistance breeding and gene therapy. However, few reports have been published that explore the efficacy of Bombyx mori nucleopolyhedrovirus (BmNPV)-inducible promoters in antiviral research in the Bombyx mori (Lepidoptera). Here, we screened BmNPV promoters (VP1054, P33, Bm21, Bm122, 39K, P143, and P6.9) and found that the 39K promoter had the highest BmNPV-induced transcriptional activity by dual-luciferase reporter assays system. By 5' truncation analysis, two regions of 39K promoter were critical for optimal virus-inducible activity, indicated that they could serve as a candidate to produce synthetic pathogen-induced promoters. Furthermore, we enhanced the virus-inducible activity of BmNPV 39K promoter using a hybrid enhancer comprising hr3 and polh-up (designated as HP39K). Finally, we showed that RNAi regulated by HP39K promoter could significantly inhibit the proliferation of BmNPV in silkworm cells. Taken together, our results have practical value in antiviral research of silkworm and baculovirus expression system.

KEYWORDS:

39K promoter; Antiviral; Baculovirus promoters; BmNPV; Silkworm; Virus-inducible promoter

PMID:
27245143
DOI:
10.1016/j.jbiotec.2016.05.037
[Indexed for MEDLINE]

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