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白蜡虫四个热休克蛋白基因的克隆与表达分析(同翅目:蚧科)

Cloning and expression analysis of four heat shock protein genes in Ericerus pela (Homoptera: Coccidae).

作者信息

Liu Wei-Wei, Yang Pu, Chen Xiao-Ming, Xu Dong-Li, Hu Yan-Hong

机构信息

*These authors contributed equally to this work.

Research Institute of Resources Insects, Chinese Academy of Forestry, Key laboratory of Cultivating and Utilization of Resources Insects of State Forestry Administration, Kunming 650224, China*These authors contributed equally to this work

出版信息

J Insect Sci. 2014 Oct 15;14. doi: 10.1093/jisesa/ieu032. Print 2014.

Abstract

To explore the function of small heat shock protein genes (shsps) and hsp70 in Ericerus pela, we cloned the full-length cDNA sequences of hsp21.5, hsp21.7, hsp70, and hsc70 and the genomic sequence of hsc70. Open reading frames of the four hsps were 570, 564, 1,908, and 1,962 base pairs (bp), respectively, which encode proteins with calculated molecular mass of 21.5, 21.7, 69.8, and 71.6 kDa. Amino acid sequence analysis revealed the presence of the conserved Hsp motifs in all four proteins. The genomic DNA of hsc70 had four introns. ep-hsp21.5 was orthologous and ep-hsp21.7 was species specific. Expression of all four transcripts during heat or cold stress and development was examined by quantitative real-time polymerase chain reaction. All four hsps were upregulated during heat or cold stress in female adults, indicating a correlation between the four hsps and heat or cold-stress tolerance in female adults. ep-hsp21.7 and ep-hsp70 were upregulated during heat stress in male larvae, implying a correlation between the two hsps and heat-stress tolerance in male larvae. The four ep-hsps were also upregulated during the developmental process in males, and ep-hsp21.5, ep-hsp70, and ep-hsc70 were upregulated in females, which indicates their possible role in the developmental regulation of E. pela.

摘要

为了探究小热休克蛋白基因(shsps)和热休克蛋白70(hsp70)在白蜡虫中的功能,我们克隆了hsp21.5、hsp21.7、hsp70和hsc70的全长cDNA序列以及hsc70的基因组序列。这四个热休克蛋白的开放阅读框分别为570、564、1908和1962个碱基对(bp),它们编码的蛋白质计算分子量分别为21.5、21.7、69.8和71.6 kDa。氨基酸序列分析显示,这四种蛋白质中均存在保守的热休克蛋白基序。hsc70的基因组DNA有四个内含子。ep-hsp21.5是直系同源的,而ep-hsp21.7是物种特异性的。通过定量实时聚合酶链反应检测了这四种转录本在热或冷应激以及发育过程中的表达情况。在成年雌虫的热或冷应激期间,所有四种热休克蛋白均上调,表明这四种热休克蛋白与成年雌虫的热或冷应激耐受性之间存在相关性。在雄性幼虫的热应激期间,ep-hsp21.7和ep-hsp70上调,这意味着这两种热休克蛋白与雄性幼虫的热应激耐受性之间存在相关性。这四种ep-热休克蛋白在雄性发育过程中也上调,而ep-hsp21.5、ep-hsp70和ep-hsc7

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8968/5443611/5b5b5e3f3b12/ieu032f1p.jpg

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