【摘要】：利用RT-PCR及RACE技術(shù),克隆到蝴蝶蘭1個幾丁質(zhì)酶基因PhCHT(GenBank登錄號為KT992851),該基因cDNA全長1 210bp,包含37bp的5′-UTR、933bp開放閱讀框和240bp 3′-UTR,編碼310個氨基酸;該蛋白為糖苷水解酶第19家族成員,兼具有溶菌酶活性;生物信息學分析顯示,該蛋白具N-端信號肽和跨膜結(jié)構(gòu),為胞外分泌蛋白;該蛋白與海棗、谷子、油棕和擬南芥的幾丁質(zhì)酶類似蛋白相近,并且在系統(tǒng)進化樹上與甘蔗和陸地棉的Ⅶ類幾丁質(zhì)酶同屬一個分支。PhCHT基因的表達分析表明,PhCHT在蝴蝶蘭營養(yǎng)器官和生殖器官中均有表達,根中表達量最高;13℃/8℃低溫處理3、6、9和15d時該基因的表達被抑制,4℃低溫處理1、2和4h表達量升高。研究表明,PhCHT基因能夠響應(yīng)短期的冷脅迫。研究結(jié)果為進一步研究蝴蝶蘭幾丁質(zhì)酶的系統(tǒng)進化及抗性育種奠定了基礎(chǔ)。
[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，

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