کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2432141 | 1106783 | 2012 | 6 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Overexpression of Hsp90 from grass carp (Ctenopharyngodon idella) increases thermal protection against heat stress Overexpression of Hsp90 from grass carp (Ctenopharyngodon idella) increases thermal protection against heat stress](/preview/png/2432141.png)
With homologous DNA probes, we had screened a grass carp heat shock protein 90 gene (CiHsp90). The full sequence of CiHsp90 cDNA was 2793 bp, which could code a 798 amino acids peptide. The phylogenetic analysis demonstrated that CiHsp90 shared the high homology with Zebrafish Grp94. Quantitative RT-PCR analysis showed that CiHsp90 was ubiquitously expressed at lower levels in all detected tissues and up-regulated after heat shock at 34 °C or cold stress at 4 °C. To understand the function of CiHsp90 involving in thermal protection, an expression vector containing coding region cDNA was expressed in E. coli BL21 (DE3) plysS. Upon transfer from 37 °C to 42 °C, these cells that accumulated CiHsp90 peptides displayed greater thermoresistance than the control cells. While incubated at 4 °C for different periods, it could also improve the cell viability. After transient transfected recombinant plasmid pcDNA3.1/CiHsp90 into mouse myeloma cell line SP2/0, we found that CiHsp90 could contribute to protecting cells against both thermal and cold extremes. On the contrary, the mutant construct ▵N-CiHsp90 (256–798 aa) could abolish the protection activity both in prokaryotic cells and eukaryotic cells. Additionally, both CiHsp90 and ▵N-CiHsp90 peptides could reduce the level of citrate synthase aggregation at the high temperature.
► CiHsp90 was up-regulated after temperature stress in all detected tissues.
► CiHsp90 could contribute to protecting cells against both thermal and cold extremes.
► The full-length CiHsp90 was required for its protection activity.
► CiHsp90 reduce the level of citrate synthase aggregation at high temperature.
► ▵N-CiHsp90 reduce the level of citrate synthase aggregation at high temperature.
Journal: Fish & Shellfish Immunology - Volume 33, Issue 1, July 2012, Pages 42–47