凡纳滨对虾内质网膜转运通道蛋白基因Sec61γ的克隆及表达分析

以生长性状为指标分离凡纳滨对虾Litopenaeus vannamei家系的肌肉组织,构建凡纳滨对虾生长性状差减cDNA文库,进行斑点杂交筛选获得内质网膜转运通道蛋白基因Sec61γ基因,克隆获得cDNA全长编码序列及部分非编码序列。序列包含270 bp的开放阅读框,推测编码的蛋白质为89个氨基酸,预测的相对分子质量为10 300,理论等电点为10.58。通过荧光定量PCR的方法研究了Sec61γ基因在对虾不同组织和不同生长期肌肉组织中的表达情况,结果表明：Sec61γ基因在对虾的血液以及心脏、肝胰腺、肌肉、胃、肠、眼柄等器官或组织中都有表达,其中在肌肉组织中表达量最高,在肠中表达量最低;在对虾不同生长期肌肉中的表达量各不相同,在10日龄的对虾肌肉组织中表达量最高,为45日龄的对虾肌肉组织中表达量的27.6倍。

Cloning and expression of Pacific white leg shrimp Litopenaeus vannamei Sec61γ gene encoding endosplasmic reticulum membrane translocation channel protein

A subtracted cDNA libraries of Pacific white leg shrimp Litopenaeus vannamei on growth was constructed,and Sec61γ gene of the shrimp was obtained by dot blot and cloning.There was a 270 bp open reading frame（ORF） encoding 89 amino acids（aa） in the sequence whose molecular weight was predicted to be 10 300 with theoretical isoelectric point of 10.58.Quantitative real time PCR was used to assess the mRNA expression of Sec61γ in different tissue and different growth period.The expression was detected in the heart,hepatopancreas,muscles,stomach,bowel,eye hilt and blood.The maximal expression level was observed in the muscle,while the minimal in hepatopancreas;the expression of Sec61γ in different growth period showed that the maximal expression level in muscles was found in 10 day old shrimp,while the minimal was found in the 45 day old shrimp.