凡纳滨对虾HSP10基因序列与低温表达分析

摘要

[Objective]HSP10 gene was cloned and studied to provide references for its regulation function in cold-resistance duration.[Method]EST data mining and RACE-PCR were used to get the whole length eDNA of LvHSP10,and bio-information methods were applied to analyze the cDNA.LvHSP10 mRNA expression in different tissues under cold stress were studied using real-time PCR.[Result]The LvHSP10 cDNA was 720 bp long containing a 306 bp ORF which encoded a 102 Aa peptide.Phylogenetic tree based on LvHSP10 amino acid sequences could reflect the evolution relations among various varieties.Real-time PCR results showed that LvHSP10 mRNA preferably expressed in all tissues and mainly expressed in muscle.According to the results from low-temperature expression profiles analysis,the LvHSP10 mRNA was down regulated in all tissues of Litopenaeus vannmei,and when the temperature descended to 13℃,its expression was the lowest.[Conclusion]LvHSP10 in Litopenaeus vannmei was conserved in gene structure and evolution relationship,and its mRNA,which was on the downside in cold stress,might negatively regulate in cold tolerance.%[目的]克隆分析凡纳滨对虾HSP10基因,为研究其在寒冷耐受过程中发挥调控功能提供参考依据.[方法]搜索EST库获得凡纳滨对虾HSP10基因序列,再通过RACE-PCR扩增获得凡纳滨对虾HSP10基因cDNA全长序列,对其进行生物信息学分析,并通过荧光定量PCR分析HSP10基因的组织表达及低温胁迫下表达量的变化.[结果]凡纳滨对虾HSP1基因cDNA全长720 bp,包含306 bp的开放阅读框,编码102个氨基酸.以各物种的HSP10蛋白序列构建系统进化树,能较好反映各物种间的进化关系.荧光定量PCR分析结果显示,凡纳滨对虾HSP10mRNA在各组织中呈遍在表达,其中以肌肉中的表达量最高.低温表达谱分析结果显示,凡纳滨对虾HSP10mRNA在低温处理凡纳滨对虾的各组织中均呈下调表达,当处理温度降至13℃,其在肝胰腺中表达量降至最低.[结论]凡纳滨对虾HSP10在结构和进化上较保守,其mRNA表达量在低温胁迫下呈下降趋势,可能在寒冷耐受过程中发挥负调控作用.