光滑鳖甲热休克蛋白70基因的克隆及表达

Cloning and expression analysis of heat shock protein 70 gene from Anatolica polita borealis(Coleoptera: Tenebrionidae)

根据GenBank中发表的昆虫热休克蛋白70(heat shock protein 70,HSP70)基因的保守序列设计引物,利用PCR结合RACE扩增的方法,获得新疆荒漠昆虫光滑鳖甲Anatolica polita borealis热休克蛋白70基因,命名为APhsp70(GenBank注册号为EF569673)。测序结果表明,序列长为2 092 bp,由核酸序列推演出的蛋白质分子量为70 kD,含653个氨基酸残基。基因结构分析表明APhsp70不含内含子序列。通过Northern blotting和半定量RT-PCR技术研究昆虫在受到不同温度胁迫时该基因的表达规律,结果表明:在45℃处理时,昆虫体内该基因的表达水平显著上升。并且,当机体受到-5℃低温胁迫并在室温恢复15 min后APhsp70的表达量也显著升高;但是4℃和-10℃低温处理组APhsp70的表达未见明显变化。

The heat shock protein 70 gene （hsp70） from Anatolica polita borealis （a desert beetle distributed in Xinjiang） was isolated through PCR and RT-PCR technique. Sequence analysis indicated that the APhsp70 gene （GenBank accession no. EF569673） has 2 092 bp in length, includes one full open reading fragment and potentially encodes a 653 amino acid residuum protein. The protein deduced from the isolated gene had a calculated molecular mass of 70 kD and was therefore named APHSP70. The APhsp70 gene contains no intron. The expression pattern of APhsp70 was examined at the transcription level after treatment with heat and cold stress. The Northern blot and semi-quantitative RT-PCR results revealed that the APhsp70 was an inducible 70 kDa isoform whose expression was specifically induced by the different stress factors.