| 摘要: |
| 采用cDNA末端快速克隆(Rapid amplification of cDNA ends,RACE)技术,以甘蔗未成熟茎cDNA为模板克隆转化酶抑制子,命名为SoInvInh1,GenBank登录号KF575175.SoInvInh1基因的cDNA序列全长为678 bp,3'-UTR长146 bp.开放阅读框长531 bp,编码176个氨基酸,预测分子质量和等电点分别为18.09 ku和8.52.SoInvInh1不含内含子序列.预测该蛋白N端具有1个信号肽和跨膜结构,19—172位氨基酸是PMEI蛋白的保守结构域.不同物种间InvInh蛋白氨基酸序列同源性较低,但都具有4个保守的Cys位点.荧光定量PCR分析表明在甘蔗茎、叶、花序和花序轴中都能检测到SoInvInh1基因表达.在甘蔗生长的不同时期,SoInvInh1在叶中表达无明显规律.而在工艺成熟期和生理成熟前期SoInvInh1整体上表现为幼茎中基因表达量高,而老茎中基因表达量低,表明茎中该基因可能与酸性转化酶活性相关. |
| 关键词: 甘蔗 转化酶抑制子 克隆 基因表达 |
| DOI：10.11841/j.issn.1007-4333.2015.03.06 |
| 投稿时间：2014-10-16 |
| 基金项目:国家"863"计划课题(2013AA102604); 国家自然科学基金项目(31360293); 国家国际合作项目(2013DFA31600); 广西自然科学基金项目(2013NXNSFAA019073); 广西八桂学者和特聘专家专项经费(2013) |
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| Cloning and expression analysis of sugarcane invertase inhibitor (SoInvInh1) gene |
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NIU Jun-qi1, HUANG Jing-li1, ZHANG Kun-kun1, YANG Li-tao1,2, LI Yang-rui1,2
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| (1.Agricultural College/State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530005, China;2.Sugarcane Research Center/Sugarcane Research Institute of Guangxi Academy of Agricultural Sciences/Key Laboratory ofSugarcane Biotechnology and Genetic Improvement (Guangxi) of Ministry of Agriculture/Guangxi Key Laboratory ofSugarcane Genetic Improvement, Chinese Academy of Agricultural Sciences, Nanning 530007, China) |
| Abstract: |
| The invertase inhibitor gene from sugarcane immature internodes was cloned using rapid amplification of cDNA ends (RACE-PCR) technique in the present study,which was named as SoInvInh1 gene with the GenBank accession number KF575175.The full-length cDNA of SoInvInh1 was 678 bp,with a 146 bp 3'-UTR.The open reading frame (ORF) was 531 bp,encoding 176 amino acids with predicted molecular weight of 18.09 ku and isoelectric point of 8.52.There was no intron in SoInvInh1.It had been predicted that there were a signal peptide and a transmembrane structure in the N-end of SoInvInh1 protein,and the region of 19-172 amino acid was a conservative domain of a PMEI protein.Though the homology of InvInh protein amino acid sequence was very low among different species,there were four conservative Cys in all of them.The results of quantitative real-time PCR (qRT-PCR) showed that the expression of SoInvInh1 could be detected in the stalks,leaves,flowers and rachis.However,the gene expression was different with different organs and growth periods.There was no obvious regularity of the gene expression in leaves in different growth periods.The SoInvInh1 showed the highest expression in immature internodes and the lowest in matured internodes at technical maturing stages and early physiological maturing stages,suggesting that the SoInvInh1 gene expression may be correlated with the activity of acid invertase in the stalk of sugarcane. |
| Key words: sugarcane invertase inhibitor gene cloning gene expression |
