Cloning and nucleotide sequencing of three heat shock protein genes (hsp90, hsc70, and hsp19.5) from the diamondback moth, Plutella xylostella (L.) and their expression in relation to developmental stage and temperature.
Cloning and nucleotide sequencing of three heat shock protein genes (hsp90, hsc70, and hsp19.5) from the diamondback moth, Plutella xylostella (L.) and their expression in relation to developmental stage and temperature.
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DOI：
10.1002/arch.20124
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发表时间：
2006-06
影响因子：
2.2
通讯作者：
S. Sonoda;M. Ashfaq;H. Tsumuki
中科院分区：
文献类型：
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作者：
S. Sonoda;M. Ashfaq;H. Tsumuki
利用RT-PCR和RACE技术，从小菜蛾（Plutella xylostella， L.）中克隆了热休克蛋白基因hsp90、hsc70和hsp19.5，并进行了测序。hsp90和hsp19.5的cDNA序列分析显示，开放阅读框（orf）长度分别为2151和522 bp，编码的蛋白分子量分别为82.4和19.5 kDa。对hsc70的cDNA进行分析，发现全长1878 bp的ORF编码一个分子量为69.3 kDa的蛋白。此外，hsc70的基因组DNA分析证实了内含子的存在，而hsp90和hsp19.5没有发现内含子。Southern blot分析表明，DBM基因组中存在每个基因家族的多个拷贝。在蛹期检测到hsp19.5的表达，在蛹期和成虫期检测到hsp90和hsc70的表达。成虫期，雌虫hsp90和hsc70的表达高于雄虫。在两性暴露于高温后，观察到这三个基因的表达均增加。这些结果表明，在DBM的发育过程中，除了热休克反应外，这些HSP基因可能还参与了其他功能。
Heat shock protein genes, hsp90, hsc70, and hsp19.5, were cloned and sequenced from the diamondback moth, Plutella xylostella (L.) by RT-PCR and RACE method. The cDNA sequence analysis of hsp90 and hsp19.5 revealed open reading frames (ORFs) of 2,151 and 522 bp in length, which encode proteins with calculated molecular weights of 82.4 and 19.5 kDa, respectively. Analysis of cDNA from hsc70 revealed an ORF of 1,878 bp coding a protein with a calculated molecular weight of 69.3 kDa. Furthermore, the analysis of genomic DNA from hsc70 confirmed the presence of introns while no introns were apparent in hsp90 and hsp19.5. Southern blot analysis suggested the presence of multiple copies of each gene family in the DBM genome. Detectable expression of hsp19.5 was observed at the pupal stage while expression of hsp90 and hsc70 was detected at both pupal and adult stages. At adult stage, females showed a higher expression of hsp90 and hsc70 than males. An increased expression was observed in all three genes after exposure to a high temperature in both sexes. These results suggest that in addition to a heat shock response, these HSP genes might be involved in other functions during the course of development in DBM.