摘要
【目的】克隆甘蔗转化酶抑制子(Invertase inhibitor)基因(SoInvInh2)的c DNA全长序列,分析基因表达模式和生物信息学特性,为该基因功能的进一步鉴定提供依据。【方法】基于同源序列克隆InvInh2基因5'序列,采用Tail-PCR技术扩增基因3'序列。采用生物信息学软件对其编码蛋白的理化性质、亚细胞定位和系统进化等进行分析。采用荧光定量PCR(qRT-PCR)分析SoInv Inh2基因在甘蔗不同生育期的组织表达特性。【结果】从甘蔗GT28幼茎中克隆到的一个新的转化酶抑制子基因,命名为SoInv Inh2,GenBank登录号KF575170。该基因的c DNA序列全长为927 bp,开放阅读框长651 bp,编码216个氨基酸。SoInvInh2基因不含内含子序列。SoInvInh2蛋白有4个不对称的α-螺旋和4个保守的Cys位点,属于InvI/PMEI家族成员。在甘蔗叶、茎、花序和花序轴中都能检测到SoInvInh2基因表达。在甘蔗不同生育期,So Inv Inh2基因在不同成熟度的叶和茎中的表达模式无明显规律。【结论】成功克隆到甘蔗SoInvInh2基因,并进行相关生物信息学分析,为该基因的进一步功能研究奠定了基础。
[Objective] The objective of this study is to clone the full-length eDNA sequence of invertase inhibitor gene （Solnslnh2） from sugarcane, and analyze its sequence and expression patterns for further identifying the biological functions of SnInvInh2. [ Method ] The 5 （ partial eDNA sequences of SnInvInh2 gene was cloned from sugarcane immature internodes using homologous sequence, and 3 （ partial eDNA sequences was cloned through Tail-PCR （Thermal asymmetric interlaced PCR）. Bioinformatic method was deployed to analyze the deduced amino acid sequence of SnInvInh2 and the physicochemical properties, subcellular localization of SnInvInh2. Phylogenetic tree was also con- structed. The spatial-specific expression of SnInvInh2 gene were determined by real-time fluorescence quantitative PCR （qRT-PCR） in dif- ferent tissue at dirt）rent growth stage of sugarcane. [ Result] A novel lnvlnh gene was cloned from the sugarcane immature intemodes, which was named as SnInvInh2 gene with the GenBank accession number KF575170. The full-length eDNA of SnInvInh2 was 927 bp, containing a 651 bp open reading frame （ORF） which encodes 216 amino acids. There was no intron in SnInvInh2. The SoInvInh2 protein contains four asymmetric a-helixes and four conservative Cys sites and belongs to the InvI/PMEI（ Invertase and pectin methylesterase inhibitory） fantily. The results of qRT-PCR sbowed that SnInvInh2 could be de- tected in the leaves, intemodes, rachis and inflorescence. At different growtb stages of sugarcane, the expression pattern of SnInvInh2 gene in leaves and steins of different maturity had no obvious regu- larity. [ Conclusion ] The successful isolation and bioinformatics anal- ysis of SnInvInh2 gene will lay tbe foundation for a furtber study.
作者
牛俊奇
黄金容
苗小荣
王道波
杨丽涛
李杨瑞
NIU Jun-qi;HUANG Jin-mng;MIAO Xiao-rong;WANG Dao-bo;YANG Li-tao;LI Yang-mi(College of Biology and Pharmacy,Yulin Normal University,Guangxi Yulin 537000,China;Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology,Guangxi Yulin 537000,China;Agricultural College/State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources,Guangxi University,Guangxi Nanning 530005,China;Sugarcane Research Center of Chinese Academy of Agricultural Sciences/Sugarcane Research Institute of Guangxi Academy of Agricultural Sciences,Guangxi Nanning 530007,China)
出处
《西南农业学报》
CSCD
北大核心
2018年第11期2239-2247,共9页
Southwest China Journal of Agricultural Sciences
基金
国家地区基金项目(31860403、51469003)
广西自然科学基金项目(2017GXNSFBA198023)
玉林师范学院高层次人才科研启动基金项目((
2017006)
玉林师范学院校级科研项目(2018YJKY27)
广西八桂学者和特聘专家专项经费(2013年)
