虹鳟鱼细胞中热休克蛋白家族的定量mRNA表达谱分析
Quantitative mRNA expression profiling of heat-shock protein families in rainbow trout cells.
作者信息
Ojima Nobuhiko, Yamashita Michiaki, Watabe Shugo
机构信息
Aquatic Genomics Research Center, National Research Institute of Fisheries Science, Fisheries Research Agency, Yokohama 236-8648, Japan.
出版信息
Biochem Biophys Res Commun. 2005 Apr 1;329(1):51-7. doi: 10.1016/j.bbrc.2005.01.097.
We isolated multiple HSPs from rainbow trout Oncorhynchus mykiss RTG-2 cells and quantitatively compared their mRNA levels between unstressed and heat-shocked cells using real-time RT-PCR analysis. Consequently, we isolated nine cDNAs encoding HSPs from heat-shocked RTG-2 cells, namely, Hsp90betaa, Hsp90betab, Grp78, Hsp70a, Hsc70a, Hsc70b, Cct8, Hsp47, and DnaJ homolog. Quantitative RT-PCR analyses, in which Hsp70b isolated previously was included, showed that the mRNA accumulation levels of Hsp70a, Hsp70b, Hsc70a, Hsc70b, and Hsp47 were significantly increased after heat shock, and the increased levels of two Hsp70s, Hsp70a, and Hsp70b, were most conspicuous. In the case of Hsc70s, the increased level of Hsc70b was more remarkable than that of Hsc70a. These results demonstrate the importance of a comprehensive expression analysis of HSPs for better understanding of the cellular stress response in fish, especially in tetraploid species such as rainbow trout.
我们从虹鳟Oncorhynchus mykiss RTG - 2细胞中分离出多种热休克蛋白（HSPs），并使用实时逆转录聚合酶链反应（RT - PCR）分析定量比较了未受应激和热休克处理的细胞中它们的mRNA水平。因此，我们从热休克处理的RTG - 2细胞中分离出9个编码HSPs的cDNA，即Hsp90betaa、Hsp90betab、葡萄糖调节蛋白78（Grp78）、Hsp70a、热休克同源蛋白70a（Hsc70a）、Hsc70b、TCP - 1环状复合物8（Cct8）、Hsp47和DnaJ同源物。定量RT - PCR分析（其中包括先前分离出的Hsp70b）表明，热休克后Hsp70a、Hsp70b、Hsc70a、Hsc70b和Hsp47的mRNA积累水平显著增加，且两种Hsp70s（Hsp70a和Hsp70b）的增加水平最为明显。就Hsc70s而言，Hsc70b的增加水平比Hsc70a更显著。这些结果表明，对HSPs进行全面的表达分析对于更好地理解鱼类，尤其是像虹鳟这样的四倍体物种中的细胞应激反应具有重要意义。
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