斑鳜肌球蛋白轻链2基因cDNA的克隆及其纵向表达分析

Cloning and Longitudinal Expression Analysis of the MCL2 Gene in Siniperca scherzeri

从斑鳜(Siniperca scherzeri)背部白色肌肉组织中提取总RNA,利用RT-PCR法克隆到斑鳜肌球蛋白轻链2基因(MLC2)(GenBank:GQ283000).斑鳜MLC2基因cDNA序列的开放阅读框的长度为513 bp,编码170个氨基酸,理论相对分子质量为19 105.61 Da,等电点为4.73.该开放阅读框具有4个EF-手相结构.斑鳜MLC2推导的氨基酸序列与已报道的其它6种鱼类MLC2氨基酸序列同源性在89%以上,其中与Ca2+结合的区域非常保守,氨基酸序列同源性为100%.用实时荧光定量PCR对斑鳜MLC2纵向表达分析显示,MLC2在斑鳜背肌的前部、中部和后部都有表达,但无显著差异.

The total RNA was isolated from the white muscle of the Siniperca scherzeri.The full length cDNA of myosin light chain 2（MCL2） was cloned using RT-PCR technology.The sequence of the MCL2 ORF is 513 bp,and encodes 170 amino acids,which includes four EF-hand structures.The putative protein of the MLC2 shows predicted relative molecular mass of 19 105.61 Da with a theoretical pI of 4.73.Compared with the MLC2 of other six fishes reported,the homology of the deduced amino acids sequences was above 89%.The primary structure of the Ca2＋ binding domain was well conserved among the MLC2s of seven fish species.The longitudinal expression analysis by real-time PCR showed that the MLC2 mRNA was expressed along longitudinal axis with some variation among the three muscle positions.