Li C H, Su X R, Wang M Q, Li Y Y, Li Y, Zhou J, Li T W
School of Marine Science, Ningbo University, Ningbo, Zhejiang Province, China.
Genet Mol Res. 2012 Aug 16;11(3):2641-51. doi: 10.4238/2012.July.10.14.
The full-length complementary DNA (cDNA) of heat shock protein 90 was cloned from Phascolosoma esculenta (PeHSP90) using expressed sequence tag and rapid amplification of cDNA end approaches. The cDNA of PeHSP90 was 2521 bp including a 5'-untranslated region of 110 bp, a 3'-untranslated region of 230 bp, and an open reading frame of 2181 bp. All of the characteristic motifs of the HSP90 family were completely conserved in the deduced amino acid of PeHSP90. The expression of PeHSP90 was induced by 3 heavy metals or elevated temperature, under which Zn²⁺ displayed effects were more toxic than those of Cd²⁺ and Cu²⁺. The polyclonal antibodies generated from the recombinant product of PeHSP90 were specifically identified not only in the recombinant product but also in the native protein from hemocytes. These results strongly suggested that PeHSP90 was involved in heavy metal challenge and thermal stress regulation in P. esculenta.
利用表达序列标签和cDNA末端快速扩增技术,从可口革囊星虫(PeHSP90)中克隆了热休克蛋白90的全长互补DNA(cDNA)。PeHSP90的cDNA为2521 bp,包括110 bp的5'-非翻译区、230 bp的3'-非翻译区和2181 bp的开放阅读框。HSP90家族的所有特征基序在PeHSP90的推导氨基酸中完全保守。PeHSP90的表达受3种重金属或高温诱导,其中Zn²⁺的毒性作用比Cd²⁺和Cu²⁺更强。由PeHSP90重组产物产生的多克隆抗体不仅能特异性识别重组产物,还能识别血细胞中的天然蛋白。这些结果强烈表明,PeHSP90参与了可口革囊星虫对重金属刺激和热应激的调节。