杧果低分子量热激蛋白基因MiHSP17.6的克隆及表达分析

Cloning and Expression Analysis of a Small Heat Shock Protein Gene(MiHSP17.6)from Mangifera indica

采用RT-PCR与RACE相结合的方法获得了杧果低分子量热激蛋白基因的c DNA全长序列,命名为Mi HSP17.6,Gen Bank登录号为KJ459857。序列分析显示,该基因序列全长为680 bp,其中开放阅读框为462 bp,编码154个氨基酸,5′非编码区和3′非编码区长度分别为80 bp和135 bp。杧果Mi HSP17.6与其他物种的低分子量热激蛋白同源性介于74%~82%。实时荧光定量分析表明,Mi HSP17.6在‘四季杧’不同组织器官中均表达,在果实发育的中期表达水平持续上升。高温(44℃)、低温(4℃)、盐(Na Cl)、聚乙二醇(PEG)、脱落酸(ABA)、双氧水(H2O2)和水杨酸(SA)处理均诱导该基因表达,因此推测Mi HSP17.6的功能可能与杧果果实发育和抵御逆境胁迫相关。

The full length sequence of c DNA,small heat shock protein 17.6 gene of mango was cloned by RT-PCR and RACE method,which was named Mi HSP17.6 with the Gen Bank accession number of KJ459857. The results showed that the full-length sequence of Mi HSP17.6 is 680 bp and the open reading frame is 462 bp,which encoding a polypeptide of 154 amino acids. The untranslated region(UTR)5′ and 3′ with the length of 80 bp and 135 bp,respectively. Comparison of the amino acids sequences of homologous HSP proteins from other species indicated that Mi HSP17.6 has a range of 74% to 82% identity. The real-time quantitative PCR results showed that Mi HSP17.6 was expressed in all tested organs. And we also found that Mi HSP17.6 was regulated by several treatments,such as high temperature(44 ℃),low temperature(4 ℃),salt(Na Cl),polyethylene glycol(PEG),abscisic acid(ABA),hydrogen peroxide(H2O2)and salicylic acid(SA)treatments. These results showed that the function of Mi HSP17.6 gene has an important role in mango fruit development and stress responses.