拟穴青蟹β-肌动蛋白基因的克隆与分析

Cloning and Analysis of β-actin Gene from Scylla paramamosain

β-肌动蛋白广泛存在于真核生物中,在维持细胞结构、细胞运动和细胞分裂等生理活动中发挥着重要作用.运用RACE技术克隆了拟穴青蟹(Scylla paramamosain)β-肌动蛋白基因,并用RT-PCR方法检测该基因在成体各组织中的表达情况.拟穴青蟹β-肌动蛋白cDNA全长1 337bp,5′端非编码区为67bp,3′端非编码区为139bp,开放阅读框1 131bp编码376个氨基酸.拟穴青蟹β-肌动蛋白与其他节肢动物β-肌动蛋白氨基酸序列的相似性高达98%～99%.系统进化树显示拟穴青蟹β-肌动蛋白基因的分子进化地位与其生物学分类地位一致.半定量RT-PCR分析结果表明,β-肌动蛋白基因在拟穴青蟹视神经节、脑神经节、胸神经节、性腺、鳃、心、胃、肌肉、肝胰腺共9个组织器官中的表达基本一致,具有良好的稳定性.

β-actin plays important roles in maintaining cytoskeletal structure,cell motility and cell division in eukaryote.In this paper,β-actin gene was cloned from Scylla paramamosain using RACE.The results showed that the full-length cDNA of β-actin gene was 1 337 bp,containing a 5′-untranslated region（5′-UTR） of 67 bp and a 3′-UTR of 139 bp.The open reading frame of 1 131 bp encodes a 376-amino acid protein.The amino acid sequence of β-actin in S.paramamosain shared high similarity with other arthropods（98%-99%）.The phylogenetic analysis was consistent with traditional classification.Semi-quantitative RT-PCR analysis showed that the expression level of β-actin gene in S.paramamosain was stable in tissues：optic ganglion,brain,thoracic ganglion,ovary,gill,heart,stomach,muscle and hepatopancreas.