飞蝗热休克蛋白70cDNA片段的克隆和序列分析

Clone and Sequence Analysis of cDNA Fragment of Heat Shock Protein 70 Gene in the Migratory Locust,Locusta migratoria

采用RT PCR方法对海南、河北和辽宁 3个飞蝗 (LocustamigratoriaL )种群的热休克蛋白 70(HSP70 )基因cDNA片段进行克隆。在事先优化的条件下 ,通过简并性上游引物和下游引物扩增出了河北种群飞蝗HSP70基因的 6 0 4bpcDNA片段 (GenBank登录号为AY2 996 37) ,推导的氨基酸序列包含 2 0 1个氨基酸残基。分析表明 ,由飞蝗该片段推导的氨基酸序列与其他昆虫的同源性较高 ;3个种群该片段的核苷酸序列相似性更高达 98 75 %。由此推测 ,飞蝗种群间抗寒性的差异可能不是HSP70的序列变异引起的 ,而与HSP70的诱导表达有关。

Using a RT-PCR method,the partial cDNA sequences of heat shock protein 70 genes from three populations (Hainan,Hebei and Liaoning of China) of migratory locusts,Locusta migratoria L.were cloned and then analyzed.Under predetermined optimal reaction conditions,one fragment of 604 bp was amplified from eggs of Hebei locust population with a pair of degenerate primers (GenBank accession No.AY299637),and deduced amino acid sequence contains 201 residues.Analyzed results show that there is high homology of amino acid sequence between the migratory locust and other insects,and nucleotide sequence identity among three locust populations is 98.75% in this fragment.Therefore,it is suggested that difference of cold hardiness between different locust populations might be connected with the induced expression of heat shock protein 70,but not the cDNA sequence divergence.