Function and Regulation Domains of a Newly Isolated Putative beta-Actin Promoter from Pacific White Shrimp

doi:10.1371/journal.pone.0122262

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	Function and Regulation Domains of a Newly Isolated Putative beta-Actin Promoter from Pacific White Shrimp
	Shi, Yingli1,2,3 ; Soderlund, Marcus 1; Xiang, Jianhai2 ; Lu, Yuanan 1
	2015-04-02
发表期刊	PLOS ONE
卷号	10 期号:4
文章类型	Article
摘要	Current development of transgenic shrimp research has been hampered due to the lack of the suitable promoters and efficient transfection methods for crustaceans. A 1642 bp sequence, containing 5'-upstream sequence, exon 1, intron 1 and partial exon 2, which is responsible for transcriptional initiation of the newly reported shrimp beta-actin (actinT1), has been isolated from the Pacific white shrimp (Litopenaeus vannamei) and named as SbaP. To determine its function and potential application in marine biotechnology, the sequence and functional domains were examined by constitutive expression of the luciferase reporter gene. We have identified 5' regions that play a central role in the expression of the mu-actin gene. The proximal promoter (-1642/-1325) contains two highly conserved transcriptional sites, CCAAT box and CArG motif. Two negative (-1140/-924, -222/-21) and one positive (-810/-425) regulatory elements have been identified in intron1. Transient transfection assay with a construct containing proximal promoter and enhancer (SbaP Delta-222/+1 Delta-1325/-924) regions of the shrimp beta-actin coupled with luciferase and EGFP (enhanced green fluorescent protein) showed that the promoter was not only functional in sf21 cells, but promoter activity was more than 8-fold higher than a viral-origin promoter (ie1, white spot syndrome virus immediate early gene promoter). Furthermore, SbaP Delta-222/+1 Delta-1325/924 drove a successful expression of luciferase injection assay in vivo injection and also showed higher promoter activity than the ie1 promoter, suggesting that the expression vectors constructed with SbaP Delta-222/+1 Delta-1325/-924 have important potential in gene transfer studies for shrimp and other crustacean species.
DOI	10.1371/journal.pone.0122262
收录类别	SCI
语种	英语
WOS记录号	WOS:000352139000090
引用统计	被引频次：5[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://ir.qdio.ac.cn/handle/337002/25457
专题	实验海洋生物学重点实验室
作者单位	1.Univ Hawaii Manoa, Dept Publ Hlth Sci, Honolulu, HI 96822 USA 2.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao, Shandong, Peoples R China 3.Univ Chinese Acad Sci, Beijing, Peoples R China
第一作者单位	中国科学院海洋研究所
推荐引用方式 GB/T 7714	Shi, Yingli,Soderlund, Marcus,Xiang, Jianhai,et al. Function and Regulation Domains of a Newly Isolated Putative beta-Actin Promoter from Pacific White Shrimp[J]. PLOS ONE,2015,10(4).
APA	Shi, Yingli,Soderlund, Marcus,Xiang, Jianhai,&Lu, Yuanan.(2015).Function and Regulation Domains of a Newly Isolated Putative beta-Actin Promoter from Pacific White Shrimp.PLOS ONE,10(4).
MLA	Shi, Yingli,et al."Function and Regulation Domains of a Newly Isolated Putative beta-Actin Promoter from Pacific White Shrimp".PLOS ONE 10.4(2015).

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