烟草糖基转移酶基因NtGT5b的克隆与序列特征分析  

Cloning and bioinformatic analysis of glucosetransferase gene NtGT5b from Nicotiana tabacum

作　　者：任学良 李立芹[2] 许力[2] 郭玉双 鲁黎明[2] 

机构地区：[1]贵州省烟草科学研究院烟草行业分子遗传重点实验室,贵州贵阳550081 [2]四川农业大学农学院,四川成都611130

出　　处：《河南农业大学学报》2016年第4期453-459,共7页Journal of Henan Agricultural University

基　　金：国家烟草专卖局科技项目(110201401007(JY-07));贵州省烟草科学研究院科技项目(烟草基因遗传转化及表型分析)

摘　　要：为研究烟草糖基转移酶的生物学功能,根据在GenBank上登录的烟草糖基转移酶基因NtGT5b(登录号BAD93690.1)的CDS序列,设计特异性引物,并以烟草栽培品种K 326总RNA逆转录后的c DNA为模板,采用PCR方法,成功获得了烟草糖基转移酶基因NtGT5b的全长CDS序列。生物信息学分析结果表明,该基因CDS全长1 458 bp,所编码的蛋白质包含485个氨基酸。其相对分子质量为54.31 k D,等电点为5.47。在二级结构上,α-螺旋和无规则卷曲为NtGT5b蛋白质的主要结构形式,而β-折叠延伸链与β-转角则相对较少。在三级结构上,该蛋白质主要由α-螺旋与β-折叠构成。NtGT5b蛋白质是一个亲水性蛋白质,含有3个糖基化位点,不含信号肽,包含有糖基转移酶家族的功能保守域PLN02410,属于GTB类型的糖基转移酶超级家族,推测具有糖基转移的功能。For the investigation of tobacco glycosyhransferase, the CDS sequence of NtGTSb （ accession number BAD93690. 1 ）, a tobacco glycosyhransferase, was applied, and specific primers were designed accordingly. For the cloning of NtGTSb, total tobacco RNA was extracted from cultivar K326 leaf, and RT-PCR was performed. Finally, full length CDS sequence of NtGT5b was successfully obtained. Bioinformatics analysis showed that the full-length CDS of NtGTSb was 1 458 bp, and the encoding protein contained 485 amino acids. Its molecular weight was 54.31 kDa, and the isoelectric point is 5.47. In the secondary structure, the alpha helix and random coil were the main structural forms of the protein, while the extended strand is relatively less. In the three-dimensional structure, NtGTSb mainly consisted of the alpha helix and β-extended strand. This protein is a hydrophilic protein, containing three glycosylation sites and does not contain signal peptides, including glycosyhransferase enzyme family function conserved domain PLN02410, belonging to the GTB type of glycosyltransferase superfamily. It is speculated that this protein has the function of glycosyhransferation.

关 键 词：烟草 糖基转移酶 基因 

分 类 号：S572[农业科学—烟草工业]