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Characterization of four heat-shock protein genes from Nile tilapia (Oreochromis niloticus) and demonstration of the inducible transcriptional activity of Hsp70 promoter

文献类型： 外文期刊

作者： Zhang, Lili 1 ; Sun, Chengfei 1 ; Ye, Xing 1 ; Zou, Shuming 2 ; Lu, Maixin 1 ; Liu, Zhigang 1 ; Tian, Yuanyuan 1 ;

作者机构： 1.Chinese Acad Fishery Sci, Key Lab Trop & Subtrop Fisheries Resource Applica, China Minist Agr, Pearl River Fisheries Res Inst, Guangzhou, Guangdong, Peoples R China

2.Shanghai Ocean Univ, Key Lab

关键词： Oreochromis niloticus;Heat-shock protein;Bacteria challenge;Streptococcus agalactia;Promoter;Transcriptional activity

期刊名称：FISH PHYSIOLOGY AND BIOCHEMISTRY （ 影响因子：2.794； 五年影响因子：2.876 ）

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收录情况： SCI

摘要： Heat-shock proteins (Hsps), known as stress proteins and extrinsic chaperones, play important roles in the folding, translocation, and refolding/degradation of proteins. In this study, we identified four Hsps in Nile tilapia (Oreochromis niloticus), which display conserved Hsp characteristics in their predicted amino acid sequences. Further analyses on the structures, homology, and phylogenetics revealed that the four Hsps belong to Hsp70 family. One of them does not contain introns and is named Hsp70, while all the other three contain introns and are named Hsc70-1, Hsc70-2, and Hsc70-3. Expressions of the four Hsp proteins were observed in all examined tissues. Six hours after infection of Streptococcus agalactiae in Nile tilapia, the expression of Hsp70 was significantly increased in the liver, head kidney, spleen and gill, while Hsc70s' expression was unchanged in all examined tissues except the head kidney that showed significantly reduced expression of both Hsc70-2 and Hsc70-3. These results suggest that Hsp70 may participate in the defense against S. agalactiae infection. We then isolated the promoter of Hsp70 gene and inserted it into the donor plasmid of Tgf2 transposon system containing green fluorescent protein (GFP) gene. The plasmid was microinjected into zebrafish embryos, where the expression of GFP was induced by heat shock, S. agalactiae immersion challenge, indicating that the isolated Hsp70 promoter has transcriptional activity and is inducible by both heat shock and bacterial challenge. This promoter may facilitate the future construction of disease-resistant transgenic fish. The work also contributes to the further study of immune response of tilapia after bacterial infection.

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