摘要
以铁观音茶树叶片为材料,利用逆转录PCR及RACE法,克隆了茶树几丁质酶基因Cs Chi(Gen Bank登录号为KR078345)。Cs Chi基因的c DNA全长为1 192 bp,包含972 bp的开放阅读框(ORF),编码323个氨基酸。生物信息学分析结果表明,Cs Chi蛋白的分子量为34.33 ku;理论等电点p I为8.44;原子组成为C1 519H2 285N413O464S18,总原子数为4 699;蛋白质结构分析显示该蛋白有6个蛋白的跨膜区域,属于跨膜蛋白;存在于细胞外;没有卷曲螺旋结构存在;Cs Chi基因编码的蛋白属于糖苷水解酶19家族,含有保守的Cht BD1结构域,与溶菌酶的保守结构域类似,可能兼具几丁质酶活性和溶菌酶活性,q PCR定量分析结果显示在不同干旱胁迫处理下茶树的Cs Chi基因的表达量,与对照组相比有所增加。推测Cs Chi基因在茶树干旱等逆境胁迫中起重要作用。
The chitinase gene CsChi （GenBank accession number： KR078345） was cloned by the RT-PCR method combined with RACE technique from the leaves of tea plant. The full length cDNA of CsChi was 1 192 bp containing an open reading frame （ORF） of 972 bp encoding 323 amino acids. Bioinformatics predicted that the molecular mass of CsChi protein was 34.33 ku. The theoretical pI was 8.44. The atomic composition was C1519H2285N413O464S18 and the total number of atoms was 4 699. The protein structure analysis showed that the protein had 6 transmembrane regions, which belonging to transmembrane protein. The protein existed the outside cell and had no coiled-coil structure. The deduced protein belonged to glycoside hydrolase family 19 and had a conserved ChtBD1 domains which had a high identity with the lysozyme, and it may be a bifunctional enzyme of chitinase and lysozyme. Analysis by qPCR showed that the transcript of CsChi under different drought stresses was up-regulated higher than the control group. We speculate that CsChi gene plays an important role in adversity stresses such as drought stress.
出处
《热带作物学报》
CSCD
北大核心
2017年第5期894-902,共9页
Chinese Journal of Tropical Crops
基金
福建省自然科学基金(No.2013J01085)
福建省重大科技专项(No.2015NZ0002-1)
福建省教育厅计划项目(No.JA15143)
关键词
茶树
几丁质酶
基因克隆
表达分析
Camellia sinensis
chitinase
gene cloning
expression analysis
