蒙古冰草肌动蛋白基因(MwACT2)克隆与表达分析

Cloning and Expression Analysis of Actin(MwACT2)in Agropyron mongolicum Keng

本研究应用RT-PCR和RACE技术,从蒙古冰草(Agropyron mongolicum Keng)中分离到1个肌动蛋白基因,命名为MwACT2。序列分析表明:MwACT2基因全长1506bp,含有1个1131bp的开放型阅读框,MwACT2蛋白长度为376个氨基酸残基,分子量为41.606KDa,理论等电点为5.29,含有actin保守位点,属于actin家族成员。该基因编码的氨基酸序列与Genebank中乌拉尔图(Triticum_urartu)肌动蛋白(登录号:EMS62134.1)氨基酸具有99%的一致性,属于非跨膜蛋白。利用半定量RT-PCR技术分析MwACT2基因的表达,结果表明该基因表达稳定,经高盐、干旱、低温处理后的表达强度无显著差异。

An Actin gene of Agropyron mongolicum Keng was isolated by RT-PCR and RACE,was named as MwACT2.Sequence analysis indicated that the full-length of MwACT2 gene is 1506 bp and contained an 1131 bp open reading frame.It encodes a polypeptide of 376 amino acids.The estimated molecular weightof the putative MwACT2 protein was 41.606 kDa with an isoelectric point of 5.29.MwACT2 has a highly conserved actin domain,which demonstrated it is a member of actin family.MwACT2 was predicted to be a non-transmembrane protein.The phylogenetic relationship also indicated that MwACT2 protein was similar to Triticumurartu（accession no.EMS62134.1）with amino acid sequences similarity of 99%.Result of semi-quantitative RT-PCR showed that the expression of MwACT2 was constant.There was no significant difference under high salt,drought and cold treatment.