杉木纤维素合成酶类似蛋白基因ClCslD1的克隆及其生物信息学分析

摘要

The protein structure of the cellulose synthase-like protein (CSL) was similar to cellulose synthase (CesA), including the conservative sequence D,D,D,QXXRW. One full-length cDNA of the cellulose synthase-like protein D (CslD) gene was cloned by reverse transcriptase (RT)-polymerase chain reaction (PCR) with 5', 3' rapid amplification of cDNA ends (RACE) methods using degenerate primers designed from the homologous sequences of the CesA genes. A multiple comparison sequence analysis was conducted concurrently with bioinformatic methods to analyze the obtained sequence. Results of the sequence analysis showed that this cDNA was 4 150 bp in length and contained a single open reading frame encoding a protein of 1132 amino acids. The multiple comparison sequence analysis showed that the deduced amino acid sequence shared high similarity (over 71%) with the ClCslD genes from Populus tremuloides, Oryza sativa, and Arabidopsis thaliana. This work will help lay an important foundation for further molecular studies with cellulose synthesis of plants.%纤维素合成酶类似蛋白(CSL)作为一种重要的膜蛋白与纤维素合成酶具有相类似的蛋白结构,都含有D,D,D,QXXRW保守区.利用其他物种中的CesA基因的保守序列设计简并引物,采用反转录-聚合酶链式反应(RTPCR)结合cDNA末端快速扩增技术(RACE)成功地从杉木Cunninghamia lanceolata中扩增出一个含有完整阅读框架的cDNA序列,长度为4 150 bp,开放阅读框架为3 396 bp,编码1 132个氨基酸并含有保守的D,D,D,QXXRW天冬氨酸残基序列.应用美国国家生物技术信息中心(NCBI)数据库对获得的基因进行序列分析比对,发现它属于CslD基因家族,命名为ClCslD1基因.多重序列比对结果分析表明,该蛋白与来自颤杨Populus tremuloides,水稻Oryza sativa,拟南芥Arabidopsis thaliana等的同源基因相似性高达71％以上.利用生物软件对其进行生物学分析,为进一步研究其在植物纤化中的功能奠定基础.