CAJ | 학술논문

运用生物信息学方法，结合RT-PCR和SMART RACE技术从烟草（Nicotiana tabacum）中克隆了4个CPI基因的全长cDNA序列，分别命名为NtCPI1、NtCPI2、NtCPI3和NtCPI4， GenBank登陆号分别为KF057988、KF057989、KF057990和KF057991。基因序列分析表明4个基因分别编码98、98、120和123个氨基酸残基的蛋白质，都具有CPI反应位点的保守基序GG、QXVXQ和A/PW，同时具有植物CPI所特有的LARFAV基序，其中NtCPI3和NtCPI4的N端还包含一段27个氨基酸残基组成的信号肽。实时荧光定量PCR试验表明，4个基因的组织表达谱很广，在根、茎、叶和芽组织中都有表达。研究结果为进一步研究半胱氨酸蛋白酶抑制剂在植物中的生理功能奠定了基础。
운용생물신식학방법，결합RT-PCR화SMART RACE기술종연초（Nicotiana tabacum）중극륭료4개CPI기인적전장cDNA서렬，분별명명위NtCPI1、NtCPI2、NtCPI3화NtCPI4， GenBank등륙호분별위KF057988、KF057989、KF057990화KF057991。기인서렬분석표명4개기인분별편마98、98、120화123개안기산잔기적단백질，도구유CPI반응위점적보수기서GG、QXVXQ화A/PW，동시구유식물CPI소특유적LARFAV기서，기중NtCPI3화NtCPI4적N단환포함일단27개안기산잔기조성적신호태。실시형광정량PCR시험표명，4개기인적조직표체보흔엄，재근、경、협화아조직중도유표체。연구결과위진일보연구반광안산단백매억제제재식물중적생리공능전정료기출。
Full-length cDNAs of fourCPI genes includingNtCPI1、NtCPI2、NtCPI3andNtCPI4were cloned fromNicotiana tabacum L. cv. K326 using RT-PCR and SMART RACE technique. Their sequences were deposited in GenBank with accession number KF057988, KF057989, KF057990 and KF057991. Sequence analysis showed that these four genes were predicted products of 98, 98, 120 and 123 amino acid residues, respectively. In addition to the typical inhibitory motifs, i.e. central signature motif QXVXG, a GG doublet in terminal region, and A/PW residues in C-terminal part. These deduced amino acid sequences contained PhyCys-specific LARFAV-like motif in the N-terminal region, of which a N-terminal signal peptide of 27 residues was found in both NtCPI3 and NtCPI4. Meanwhile, transcripts of these four genes were found in roots, stems, leaves and buds by real-time quantitative PCR, which indicated that they were broadly expressed in tobacco. This study laid foundation for further exploring physiological functions of these cysteine proteinase inhibitor genes in plants.