新疆主栽大蒜品种的蒜酶基因序列分析  

Sequence Analysis of Alliinase Genes for Varieties Garlic in Xinjiang

作　　者：刘旭新[1] 吐尔逊娜依·迪丽夏提 石强[1] 韩志敏[1] Liu Xuxin;Dilxat Tursunay;Shi Qiang;Han Zhiming(Xinjiang Agricultural Vocational Technological College,Changji,831100)

机构地区：[1]新疆农业职业技术学院,昌吉831100

出　　处：《分子植物育种》2021年第13期4278-4285,共8页Molecular Plant Breeding

基　　金：新疆农业职业技术学院科研基金(XJNZYKJ201716)资助。

摘　　要：本研究克隆得到并利用生物信息学分析4个新疆大蒜品种的蒜酶基因cDNA的序列,比较了它们之间序列差异性及其在不同葱蒜类植物中的进化关系。以新疆主栽的4个大蒜品种为实验材料,利用试剂盒利用试剂盒提取总RNA后,经RT-PCR克隆获得900 bp的蒜酶DNA片段。利用生物信息学工具,在比对分析新疆大蒜品种间蒜酶序列差异和其他地区的葱蒜类植物中发现4个品种的大蒜蒜酶cDNA具有明显的单核苷酸差异(SNP),且蛋白质的氨基酸序列同源性很高;计算实验获得的蒜酶cDNA序列得到蒜酶蛋白氨基酸序列及结构,发现新疆4种大蒜蒜酶蛋白的氨基酸序列差异较小,经计算分析同源二聚体结构后,蒜酶蛋白形成的同缘双亚基结构在体内相对稳定,说明蒜酶在蛋白的一级结构具有种间保守性;计算分析得出蒜酶蛋白序列N端有信号肽结构,其可能成为胞内转运的主要识别位点。新疆主栽大蒜蒜酶cDNA序列的研究及SNP特征为鉴定大蒜品种提供参考依据,为后续体外表达蒜酶蛋白奠定理论基础。This study is analyzing c DNA sequence of alliinase for 4 varieties of garlic grown in Xinjiang,and using bio-informatics tools to study the differences of alliinase sequences and phylogenetic relationship with other garlics.The 4 garlic varieties mainly grown in Xinjiang are experimental materials.Total RNA was extracted with the kit,and 900 bp allicase DNA fragments were cloned by RT-PCR.Using bioinformatics tools,there found significant differences(SNP)in Xinjiang 4 garlic varieties and significant evolutionary differences,we compared sequences of alliinase from Xinjiang garlic with others of different region originated garlic and onion,the homology of amino acid sequence of proteins was very high.The structer of predicted proteins of alliinase according to sequence of c DNA was calculated and analyzed,the sequences of alliinase protein for 4 garlics present very small difference in sequence.The alliinase homo-dimer protein model was built by using cDNA sequences obtained,it was a stable homo-dimer protein in vivo,shows that alliinase has interspecies conserved in the primary structure of the protein.The N-terminal of alliinase protein sequence has signal peptide structure,which may be the main recognition site of intracellular transport.The SNP of 4 cDNA sequences of alliinase was identified,which give us a method for identification of garlic varieties,this work also give us preparation for expressing functional protein of alliinase in vitro.

关 键 词：大蒜 蒜酶 RT-PCR 序列比对 SNP 

分 类 号：S633.4[农业科学—蔬菜学]