中国美利奴绵羊磷脂酶C zeta基因的克隆及序列分析

Cloning and Sequencing Analysis of PLC ζ Gene from Chinese Merino Sheep

根据GenBank中公布的牛磷脂酶C zeta(PLCζ)基因序列设计引物,通过RT-PCR方法对中国美利奴绵羊的PLCζ基因进行扩增,以了解其分子生物学特征。测序表明,美利奴绵羊PLCζ基因开放阅读框全长1 905bp,共编码634个氨基酸。生物信息学分析所扩增的中国美利奴绵羊PLCζ基因具有典型的EF-hand钙结合结构域、磷脂酰肌醇特异性磷脂酶X结构域、磷脂酰肌醇特异性磷脂酶Y结构域、蛋白激酶C2保守区域,其170和215氨基酸位点为组氨酸活性位点。与牛、猪、犬、人、猴、鼠、兔和原鸡的PLCζ基因核苷酸同源性分别为97.32%、88.19%、73.93%、81.42%、83.05%、73.65%、72.42%和65.07%。氨基酸序列的系统进化树表明中国美利奴绵羊PLCζ基因与牛的亲缘关系最近。

In order to reveal the molecular biological characteristics of phospholipase C zeta （PLC ξ） gene of Chinese Merino sheep, the PLC ξ gene was amplificated by RT-PCR using the primers designed based on bovine PLCξ sequence of in GenBank. The sequence, results showed that the complete CDS contains 1905bp in one ORF, which encodes 634 amino acids （aa）. Bioinformation analysis showed that the length of fragment is similar in structure to other PLC isoforms, PLCξ spans four EF hand domains at the N ter- minus, X and Y catalytic domains, and a C2 terminus domain, and the 170 aa and 215 aa are histidine active site. Compared with conferential PLCξ gene sequence of other animals published in GenBank, the homologies of Chinese Merino sheep and bovine, porcine, dog, human, monkey, mouse, rat and chicken are up to 97.32% ,88. 19%0,73.93% ,81.42% ,83.05% ,73.65% ,72.42% ,65.07%, respectively. Evolution analysis indicated that the PLCξ gene of Chinese Merino sheep was closely related to that of bovine.