海蓬子DnaJ-like基因片段的表达和生物信息学分析

Expression and Bioinformatics Analysis of DnaJ-like Gene Sequence from Salicornia bigelovii Torr.

从盐生植物海蓬子的抑制消减杂交文库中克隆了DnaJ-like基因的长609 bp的DNA序列,编码1个202 aa的开放阅读框,Northern B loting显示该基因在检测子(200 mmol/L NaC l)中的表达显著增强。生物信息学分析表明,在蛋白质水平上,该片段与拟南芥和水稻的同源性分别为6 e-68和1 e-58;在核酸水平上,该片段与拟南芥的同源性为3 e-24。保守区分析显示,该片段含有DnaJ-like基因完整的J功能域(J-Dom ain,66 aa),功能域在蛋白质水平与GenBank两种相关类型的J功能域的同源性分别为1 e-11和9 e-15。这为该基因的克隆及研究其在植物逆境胁迫中的作用奠定了基础。

A 609 bp DNA sequence of DnaJ-like gene was successfully cloned from halophyte, Salicornia Bigelovii Tort. ,during early stage of salt stress based on suppression subtractive hybridization （SSH） and comfirmed by BLAST x/n analysis. The result of Northern bloting indicated that the represented gene was significantly upregulatedly expressed under 200 mmoL/L NaCl treatment compared to non-NaC; treatment. Bioinformatics analysis showed that the sequence of DnaJ-like gene in halophyte has high E-values, 6e-68 and 1e-58, when compared with Arabidopsis thaliana and Oryza sativa at amino acid level（Blastx） and the E-value reached 3e-24 when compared with Arabidopsis thaliana DnaJ-like cDNA sequcnce at nucleotide level （Blastn）. Conserved domain analysis made it clear that an integrated J-domain was involved in the sequence. J-domain has E value of le-11 and 9e-15 compared with two types J-domains in GenBank CDD database. All these have widely suggested that it will be expectable to clone the full length cDNA of DnaJ-like gene from the inherent high salttolerant species.