甘蔗核苷二磷酸激酶(NDPK1)基因克隆及表达分析  

Molecular Cloning of Sugarcane NDPK1 Gene and Its Expression Analysis

作　　者：梁潘霞[1,2,3,4] 李杨瑞[1,2,4] 杨丽涛[1,4] 

机构地区：[1]广西大学亚热带农业生物资源保护与利用国家重点实验室,广西南宁530004 [2]广西作物遗传改良生物技术重点开放实验室,广西南宁530007 [3]广西农业科学院资源与环境研究所,广西南宁530007 [4]中国农业科学院甘蔗研究中心/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,广西南宁530007

出　　处：《热带作物学报》2012年第12期2199-2205,共7页Chinese Journal of Tropical Crops

基　　金：国家科技支撑计划项目(No.2007BAD30B00);科技部国际合作项目(No.2008DFA30600、2009DFA30820);广西自然科学基金团队项目(No.2011GXNSFF018002);广西科技攻关项目(桂科能No.0815011);广西农业科学院团队项目(桂农科No.2011YT01);广西农业科学院基本科研业务专项项目(No.201019)

摘　　要：采用RT-PCR技术克隆了甘蔗核苷二磷酸激酶基因全长cDNA,并用荧光定量PCR分析该基因的表达情况。结果表明:克隆得到的cDNA片段长度为686 bp,包括1个450 bp的开放阅读框,编码149个氨基酸,GenBank登录号为JQ712580。甘蔗与高粱NDPK基因cDNA序列的同源性为96%,氨基酸序列同源性为98%;该基因推导的蛋白分子量大小为16.83 ku,等电点为6.58,疏水性分值在-2.089～2.033;蛋白二级结构中α-螺旋占44.3%,随机卷曲占24.83%,延伸链占20.13%,β-转角只占10.74%。实时荧光定量分析结果表明,随着甘蔗干旱胁迫时间的延长,NDPK1基因表达均呈先升高后降低的趋势,30 h达最大;在处理后的18、30、40 h,PEG与PEG+Si处理的NDPK1基因的表达量有显著差异。In this study, the full length of NDPK1 cDNA was obtained by RT-PCR and its expression was analyzed by quantitative real-time PCR. The results showed thatNDPK1 was consisted of 686 bp with an open reading frame of 450 bp, encoding a polypeptide of 149 amino acids, and its GenBank accession number is JQ712580. Sequences alignment showed that the homology of cDNA sequence between sugarcane and sorghum bicolor NDPK1 gene was 96%, while the homology of amino acid between them was 98%. The deduced protein molecular weight was 16.83 Ku withPI 6.58 and hydrophobic from-2.089 to 2.033. The molecular structure prediction results suggested that the protein contained 44.3% α-helix, 24.83% loop, 20.13% extended strand and 60.3% β-meander. The results of quantitative real-time PCR analysis showed that the mRNA of NDPK1 was increased initially and then increased with time of PEG stress, and NDPK1 had the most abundant expression at 30 h of the treatment; Significant differences were observed between the treatments of PEG and PEG＋Si at 18, 30, 40 h of the treatment.

关 键 词：甘蔗 核苷二磷酸激酶 克隆 生物信息学分析 表达 

分 类 号：S566.1[农业科学—作物学]