| 摘要: |
| 【目的】克隆一个苹果山梨醇转运子基因（命名为ST）的全长cDNA，为进一步研究该基因的功能奠定基础。【方法】利用RT-PCR法从苹果叶片中克隆ST基因，应用相关软件对其进行生物信息学分析；并采用实时荧光定量RT-PCR(Real-time RT-PCR)法，分析ST在苹果不同组织中以及在果实发育过程中的表达情况。【结果】获得了1个包含全长开放阅读框的ST基因，该基因cDNA全长1 767 bp，包含长达1 617 bp的完整开放阅读框，该基因所编码的蛋白与GenBank中已登录的其他植物山梨醇转运子的同源性均在72%以上，其中与苹果山梨醇转运子MdSOT5的同源性较高，为81%。 Real-time RT-PCR结果表明，在不同组织中，ST在韧皮部的相对表达量最高，其次在衰老叶、成熟叶、叶柄中较高，在库器官花、幼叶、果实、果柄、根中表达较低；在果实不同发育时期，ST基因表达量在果实发育早期很低，花后30 d快速升高并达到峰值，在发育中期保持一个较高的表达水平，在果实发育后期相对表达量又降低，之后略呈递增趋势。【结论】从苹果叶片中克隆了ST基因，生物信息学分析可知ST具有糖转运功能；ST在韧皮部的表达量最高，说明其负责长距离运输；ST在果实发育过程中的表达量与山梨醇的含量变化相一致。 |
| 关键词: 苹果 山梨醇转运子 基因克隆 表达分析 |
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| 基金项目:国家“863”高新技术研究与发展计划项目（2008AA10Z157）；陕西省重大科技专项计划项目（2006kz05-G4） |
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| Cloning and expression analysis of a novel member of the sorbitol transporter family from apple（Malus domestica Borkh.） |
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| Abstract: |
| 【Objective】Genes encoding sorbitol transporter (ST) was cloned from apple leaf,which is a member of apple sorbitol transporter family,bioinformatics analysis made and the temporal and spatial expression of ST studied in different tissues and organs of apple,as a basis to further study gene function.【Method】Using RT-PCR,cloned ST from apple leaves,and bioinformatics analysis was performed with related software;Using Real-time RT-PCR,we analyzed the expression of STin different organs of apple and fruit development.【Result】We analyzed and found that ST cDNA contains a full open reading frame.The length of cDNA of ST gene is 1 767 bp,which contains a full open reading frame (1 617 bp).Homology between ST gene and sorbitol transporter of other plants logged in GenBank is over 72%,and the highest homology (81%) with apple sorbitol transporters MdSOT5.Real-time fluorescence quantitative analysis showed that ST has the highest expression in the phloem,high expression in source leaves and lower expression in the sink organs in different tissues.Also in the different developmental stages of fruit,expression of ST is very low in early fruit development,rapidly rises to peak after 30 days of flowering,maintains a high level of expression in the medium term development,and reduces in late fruit development.【Conclusion】We cloned ST from apple leaves,and showed that ST has the function of glucose transporter through bioinformatics analysis.We studied the quantitative expression patterns,and results indicated that the expression of ST is the highest in the phloem,and ST is responsible for long distance transport.In fruit development,the variation of ST is consistent with that of sorbitol. |
| Key words: apple sorbitol transporter gene cloning gene expression |
