朱砂叶螨几丁质酶TecCht2基因的克隆及特征分析

摘要

[Objective]The chitinase is critical to chitin metabolism.The aim of this study is to clone chitinase gene of Tetranychus cinnabarinusand then analyze its character.[Methods]In this study,the chitinase gene sequence of T.cinnabarinus was obtained by searching its transcriptome data and assembling the chitinase sequence fragments.According to the chitinase sequence of Tetranychus,the specific PCR primers were designed.The chitinase gene was cloned and analyzed detailedly.The SWISS MODEL was used to predict the protein tertiary structure,especially the catalytic domain.[Results] The obtained gene TecCht2,which was submitted to GeneBank (accession number KY705296),was confirmed to be the chitinase gene sequence of T.cinnabarinus by sequencing and sequences homology analysis.According to our results,the open reading frame of TecCht2 is 1875bp,encoding 624 amino acids,and its C side have a chitin binding domain.TecCht2 has typical structure character of chitinase.[Conclusion]The obtained chitinase gene of T.cinnabarinus laid the basis for the studies of chitinase function and its application for mite control.%[目的]拟克隆朱砂叶螨几丁质代谢中的关键基因几丁质酶基因,并对其进行特征分析.[方法]利用转录组拼接技术对朱砂叶螨转录组进行拼接,根据叶螨几丁质酶序列通过BLAST找到朱砂叶螨几丁质酶相似序列,设计PCR引物,进行朱砂叶螨几丁质酶基因克隆,分析其详细特征.利用SWISS MODEL预测其蛋白结构,特别是催化功能区的结构.[结果]测序分析确认为朱砂叶螨几丁质酶基因序列,并提交到GeneBank(KY705296),其开放阅读框为1 875 bp,编码624个氨基酸,C端有几丁质结合区.根据新的几丁质酶分类,判定TecCht2为Ⅵ型几丁质酶,其结构具有典型的几丁质酶结构特征.[结论]本研究克隆得到的朱砂叶螨几丁质酶基因,为今后几丁质酶功能和用于害螨防治研究奠定基础.