蝴蝶兰几丁质酶基因的克隆及表达特性分析

发布时间：2018-10-20 13:09

【摘要】：利用RT-PCR及RACE技术,克隆到蝴蝶兰1个几丁质酶基因PhCHT(GenBank登录号为KT992851),该基因cDNA全长1 210bp,包含37bp的5′-UTR、933bp开放阅读框和240bp 3′-UTR,编码310个氨基酸;该蛋白为糖苷水解酶第19家族成员,兼具有溶菌酶活性;生物信息学分析显示,该蛋白具N-端信号肽和跨膜结构,为胞外分泌蛋白;该蛋白与海枣、谷子、油棕和拟南芥的几丁质酶类似蛋白相近,并且在系统进化树上与甘蔗和陆地棉的Ⅶ类几丁质酶同属一个分支。PhCHT基因的表达分析表明,PhCHT在蝴蝶兰营养器官和生殖器官中均有表达,根中表达量最高;13℃/8℃低温处理3、6、9和15d时该基因的表达被抑制,4℃低温处理1、2和4h表达量升高。研究表明,PhCHT基因能够响应短期的冷胁迫。研究结果为进一步研究蝴蝶兰几丁质酶的系统进化及抗性育种奠定了基础。
[Abstract]:Using RT-PCR and RACE techniques, a chitinase gene PhCHT (GenBank accession number of Phalaenopsis (KT992851) was cloned. The total length of cDNA gene was 1 210bp, which contained the open reading frame of 37bp and the 310-amino acid sequence of 37bp, and the protein was a member of the 19th family of glucoside hydrolase. Bioinformatics analysis showed that the protein had N-terminal signal peptide and transmembrane structure and was an extracellular secretory protein, which was similar to the chitinase similar proteins of jujube, millet, oil palm and Arabidopsis thaliana. PhCHT gene expression analysis showed that PhCHT was expressed in vegetative organs and reproductive organs of Phalaenopsis. The expression of this gene was inhibited at 13 鈩，

本文编号：2283280