金花茶CnBCH基因cDNA全长克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of the Full-length of cDNA Sequence of CnBCH Gene from Golden Camellia (Camellia nitidisima)

作　　者：周兴文[1] 周银慧 赵英[1] 朱宇林[1] Zhou Xingwen Zhou Yinhui Zhao Ying Zhu Yulin(College of Biology and Pharmacy of Yulin Normal University, Yulin, 53700)

机构地区：[1]玉林师范学院生物与制药学院,玉林537000

出　　处：《分子植物育种》2017年第1期77-83,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31360197);玉林师范学院高层次人才科研启动基金(G20120027)共同资助

摘　　要：利用CODEHOP和RACE方法,从珍稀观赏植物金花茶(Camellia nitidissima)花瓣中克隆得到了β-胡萝卜素环羟化酶(β-carotene hydroxylase,BCH)基因的cDNA全长,命名为CnBCH。碱基序列分析结果表明,该CnBCH基因全长1 089 bp,包含58 bp的5'非翻译区(untranslated regions,UTR)、105 bp的3'UTR和927 bp编码308个氨基酸的开放阅读框。生物信息学分析表明,该基因编码的蛋白质为不稳定亲水性蛋白,分子量为34.42 k D,无信号肽,含有4个跨膜结构域,一个脂肪酸羟化酶超家族(FA_hydroxylase super family)功能结构域以及BCH功能结构域(PLN02601);CnBCH二级结构以α-螺旋为主,其次为无规则卷曲,β-折叠所占比例最少。氨基酸序列比对分析结果显示,CnBCH与茄科、蔷薇科等植物BCH蛋白同源性都在70%以上,与柿树(Diospyros kaki T.)BCH同源性最高。本研究为进一步了解CnBCH基因的功能及其在金花茶花瓣类胡萝卜素合成中的作用提供了帮助。By the methods of CODEHOP and rapid amplification of cDNA ends（RACE）, full-length of cDNA sequence of β-carotene hydroxylase（BCH） gene was cloned from the petals of golden camellia（Camellia nitidissima） which is a rare and famous species of China, that was named as CnBCH. Nucleotide sequence analysis showed that the length of cDNA sequence of CnBCH is 1 089 bp, which contains 5’-untranslated region of 58 bp,3’-untranslated region of 105 bp, and an opening reading frame（ORF） of 927 bp encoding 308 predicted amino acids. Bioinformatics analyses showed that the molecular of CnBCH is 34.42 k D and no signal peptide, there are 4transmembrane domains, a FA_hydroxylase super family domain and β-carotene hydroxylase domain（PLN02601）in CnBCH. The analysis data of the second structrure of CnBCH indicated that alpha helix is the largest structural element followed by random coil and β-sheet. Sequence alignment of amino acids revealed that CnBCH shared more than 70% homology with BCH from plants of Solanaceae, Rosaceae, and the highest homology with Persimmon Tree（Diospyros kaki T.）. This research provided substantial theoretical help of furtherly studying on the gene function and the role in carotenoids biosynthesis of golden camellia（C. nitidissima） petals.

关 键 词：金花茶 Β-胡萝卜素羟化酶 克隆 

分 类 号：S685.14[农业科学—观赏园艺] Q943.2[农业科学—园艺学]