title: Characterization of four heat-shock protein genes from Nile tilapia (Oreochromis niloticus) and demonstration of the inducible transcriptional activity of Hsp70 promoter.
abstract: 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.
authors.affiliation: Key Laboratory of Tropical and Subtropical Fisheries Resource Application and Cultivation, China Ministry of Agriculture; Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Zhang
authors.forename: Chengfei
authors.initials: C
authors.lastname: Sun
authors.forename: Xing
authors.initials: X
authors.lastname: Ye
authors.forename: Shuming
authors.initials: S
authors.lastname: Zou
authors.forename: Maixin
authors.initials: M
authors.lastname: Lu
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Liu
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Tian
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D023681:Cichlids / Q000235:genetics* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D006358:Hot Temperature; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011897:Random Allocation; D017422:Sequence Analysis, DNA; D013290:Streptococcal Infections / Q000378:metabolism / Q000662:veterinary; D013292:Streptococcus agalactiae; D014158:Transcription, Genetic / Q000235:genetics*
chemical_list: D018840:HSP70 Heat-Shock Proteins
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