松墨天牛葡萄糖-6-磷酸异构酶基因克隆及序列分析

Cloning and sequence analysis of glucose-6-phosphate isomerase gene in Monochamus alteratus

【目的】克隆松墨天牛(Monochamus alteratus)葡萄糖-6-磷酸异构酶(GPI)基因c DNA序列,探究其分子特征,为深入研究松墨天牛GPI的分子功能打下基础。【方法】采用RACE技术克隆松墨天牛GPI基因c DNA序列,并对该序列及其编码氨基酸序列进行生物信息学分析。【结果】克隆并鉴定松墨天牛GPI基因,命名为Ma GPI(Gen Bank登录号:KU323592),该基因序列长1038 bp,共编码279个氨基酸。同源比对分析发现,松墨天牛与赤拟谷盗(Tribolium castaneum)的同源性最高,达90%,而与其他昆虫的同源性在76%以上。系统发育进化树分析结果表明,松墨天牛与赤拟谷盗在同一分支,与家蚕(Bombyx mori)相隔最远。Ma GPI为亲水蛋白,含有5个功能位点:活性位点、变构位点、活性部位盖子和两个多肽结合位点。【结论】明确了Ma GPI的核苷酸序列及编码蛋白特征,为进一步研究Ma GPI的分子功能提供依据。

【Objective】The present experiment was conducted to clone full-length c DNA of glucose-6-phosphate isomerase（GPI） from Monochamus alteratus, and analyze its molecular characteristics, in order to lay the foundation for further studying its functions. 【Method】The c DNA of GPI gene was cloned from M. alteratus using RACE technology. The gene sequcence and its amino acid sequence were analysed using bioinformatics softwares. 【Result】The GPI gene was cloned successfully from M. alteratus, and named Ma GPI（Gen Bank accession no. KU323592）. Its c DNA was 1038 bp in length,encoding 279 amino acids. The alignment of amino acid sequences showed that, Ma GPI shared the highest homology（90%） with GPI gene of Tribolium castaneum, and shared homology of 76% with other insects. The phylogenetic tree showed that M. alteratus and T. castaneum was in the same branch, and far from Bombyx mori. Ma GPI was a hydrophilic protein, with 5 functional sites viz., active site, allosteric site, cap of active site, peptide binding site. 【Conclusion】The sequence of nucleotide and characteristics of amino acid sequence encoded by Ma GPI gene have been clarified, in order to provide a foundation for further researching molecular function of Ma GPI.