凡纳滨对虾HSP1噬因序列与低温表达分析

【目的】克隆分析凡纳滨对虾HSPIO~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HSPIO~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HSPIO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HSP10基因cDNA全长720bp，包含306bp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HSPIOmRNA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HSPIOmRNA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。

Sequence of HSPIO gene in Litopenaeus vannmei and its low temperature expression analysis

Objective]HSPlO gene was cloned and studied to provide references for its regulation function in cold-resis- tance duration. Method]EST data mining and RACE-PCR were used to get the whole length cDNA of LvHSPIO, and bio-in- formation methods were applied to analyze the eDNA. LvHSPIO mRNA expression in different tissues under cold stress were studied using real-time PCR. Result]The LvHSPIO cDNA was 720 bp long containing a 306 bp ORF which encoded a 102 Aa peptide. Phylogenetic tree based on LvHSPIO amino acid sequences could reflect the evolution relations among various vari- eties. Real-time PCR results showed that LvHSPIO mRNA preferably expressed in all tissues and mainly expressed in muscle. According to the results from low-temperature expression profiles analysis, the LvHSPIO mRNA was down regulated in all tis- sues of Litopenaeus vannmei, and when the temperature descended to 13℃, its expression was the lowest. Conclusion]LvH- SPIO 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.