甘蔗新基因ShHXT4克隆亚细胞定位及表达特征

发布时间：2018-11-18 12:47

【摘要】：以甘蔗品种新台糖22号为材料,利用同源克隆技术从甘蔗中分离出ShHXT4基因,以GADPH基因为内参,并采用实时定量RT-PCR分析该基因在甘蔗组织中的表达,同时构建亚细胞定位载体,通过农杆菌侵染洋葱表皮的方法对ShHXT4编码的蛋白进行亚细胞定位。研究分析发现该基因CDS序列长度为1 632 bp,编码540个氨基酸,蛋白的等电点(p I)为9.16,理论分子量为56.64 k D,命名为ShHXT4,基因编码多肽链中包含信号肽和12次跨膜结构域,且亚细胞定位于质膜,编码一种转运蛋白。洋葱表皮瞬时表达该基因GFP融合载体表明,ShHXT4定位于细胞膜,组织表达分析显示ShHXT4基因在根、茎和叶中均有表达,叶片中的表达量较高,暗示ShHXT4基因可能在甘蔗叶片糖分转运过程中扮演重要角色,研究结果为进一步研究ShHXT4基因的功能及应用提供帮助。
[Abstract]:The ShHXT4 gene was isolated from sugarcane by homologous cloning technique and the GADPH gene was used as internal reference. The expression of the gene in sugarcane tissue was analyzed by real-time quantitative RT-PCR. At the same time, the subcellular localization vector was constructed, and the protein encoded by ShHXT4 was located by Agrobacterium tumefaciens infecting onion epidermis. The CDS sequence length of the gene was 1 632 bp, encoding 540 amino acids, the isoelectric point (p I) of the protein was 9.16, and the theoretical molecular weight was 56.64 KD, which was named ShHXT4,. The gene encoding polypeptide chain contains signal peptides and 12 transmembrane domains, and the subcells are located on the plasma membrane and encode a transporter. The GFP fusion vector for the transient expression of the gene in onion epidermis showed that ShHXT4 was located on the cell membrane. Tissue expression analysis showed that the ShHXT4 gene was expressed in root, stem and leaf, and the expression level in leaf was higher. It is suggested that ShHXT4 gene may play an important role in sugar transport in sugarcane leaves, and the results may be helpful to further study the function and application of ShHXT4 gene.
【作者单位】：中国热带农业科学院热带生物技术研究所中国热带农业科学院甘蔗研究中心农业部热带作物生物学与遗传资源利用重点实验室;
【基金】：海南省自然科学基金(20163124) 中央级公益科研院所基本业务费(ITBB140503) 国家自然科学基金项目(No.31371687)共同资助
【分类号】：S566.1

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