文献类型：期刊文献

中文题名：山鸡椒1-脱氧木酮糖-5-磷酸还原异构酶DXR基因的克隆和SNP分析

英文题名：Cloning and SNP Analysis of 1-Deoxy-D-xylulose-5-phosphate Reductoisomerases(DXR) in Litsea cubeba

作者：刘英冠[1] 吴庆珂[1] 何关顺[2] 汪阳东[1] 杨素素[1] 陈益存[1] 高暝[1]

第一作者：刘英冠

机构：[1]中国林业科学研究院亚热带林业研究所;[2]广西壮族自治区田林县林业局

年份：2015

卷号：25

期号：1

起止页码：93-100

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；CSCD:【CSCD2015_2016】；

基金：国家自然科学基金项目(31370576)

语种：中文

中文关键词：山鸡椒;DXR;基因克隆;SNPs

外文关键词：Litsea cubeba; DXR; gene cloning; SNPs

分类号：S718.46

摘要：在转录组测序结果分析基础上,以山鸡椒c DNA为模板,克隆得到山鸡椒1-脱氧木酮糖-5-磷酸还原异构酶DXR基因c DNA全长,以山鸡椒基因组DNA为模板,设计引物、扩增拼接后获得山鸡椒DXR基因全长,命名为Lc DXR。序列分析表明,Lc DXR c DNA全长为1 501 bp,5’非编码区长34 bp,3’非编码区长53 bp,开放阅读框长1 413 bp,预测编码含有470个氨基酸残基的蛋白质,等电点为6.62,分子量为51.12 k D。Lc DXR基因全长为12 601bp,其中外显子12个,内含子11个。对来自10个种源的Lc DXR基因编码区单核苷酸变异位点进行分析表明:在c DNA区间内共发现10个SNP(single nucleotide polymorphism)位点,其中有4个单核苷酸变异导致了所编码的氨基酸的改变,为了分析氨基酸突变导致的蛋白质精细结构的变化,利用Swiss-PDB Viewer模拟4个突变位点氨基酸残基的替换。其中江西安远(AY)的突变Lys119Thr引起了氢键的变化,推测可能对酶的活性产生影响。研究结果为深入研究山鸡椒脱氧木酮糖5-磷酸还原异构酶的活性和功能奠定了基础,同时为山鸡椒遗传育种提供理论依据。
Based on the results of transcriptome sequencing, the DXR gene cDNA was isolated from the flower bud of Litsea cubeba by the method of RT-PCR. At the same time, full-length sequence of DXR was first obtained from the genomic DNA by PCR and splice, and named as LcDXR. Sequence analysis showed that the full-length eDNA of LcDXR was 1 501bp, including 5＇ non-coding region 34bp and 3＇ non-coding region 53bp and encoded 470 amino acids. The theoretical molecular weight of LcDXR was 51.12kd and the isoelectrie point was predicted as 6.62. The full-length gene was 12 601bp with 12 exons and 11 introns. Single nucleotide polymorphic （SNP） sites of LcDXR from 10 sources were analyzed, the results showed that there were 10 SNPs within eDNA region, and 4 SNPs lead to amino acids changes. Point mutation was analyzed by Swiss-PDB Viewer to find fine structure changes resulted by amino acids mutations. The Lys119Thr mutation in Anyuan, Jiangxi Province was shown to have H-bonds change which may have an impact on the enzyme activity.