| 甜瓜果实酸性转化酶基因反义表达载体的构建(英文) |
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| 引用本文： | 于喜艳,田红梅,樊继德,王秀峰.甜瓜果实酸性转化酶基因反义表达载体的构建(英文)[J].果树学报,2006,23(6):850-853. |
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| 作者姓名： | 于喜艳 田红梅 樊继德 王秀峰 |
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| 作者单位： | 山东农业大学园艺科学与工程学院,泰安,271018 |
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| 基金项目： | 国家自然科学基金
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山东农业大学校科研和教改项目 |
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| 摘 要： | 应用RNA反义技术抑制甜瓜果实发育过程中酸性转化酶活性,促进蔗糖积累,从而为培育优质品种提供了可行的新方法。将已克隆到pMD18-T载体上的甜瓜酸性转化酶基因用BamHⅠ和HincⅡ双酶切,得到该基因编码区1038bp的cDNA片段,将其定向插入到植物表达载体pROK2的BamHⅠ/SmaⅠ克隆位点,构建了甜瓜酸性转化酶cDNA反义表达载体(Anti-MAI1)。采用冻融法将其转入根癌农杆菌LBA4404,得到了完整的Ti质粒表达载体系统。利用叶盘法转化烟草,经PCR和PCR-Southern杂交检测,证明此基因已整合入烟草的核基因组中。
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| 关 键 词： | 甜瓜 酸性转化酶基因 反义载体 烟草 转化 |
| 文章编号： | 1009-9980（2006）06-850-04 |
| 收稿时间： | 2006-03-13 |
| 修稿时间： | 2006-07-21 |
Construction and study of a vector for antisense expression of acid invertase gene from melon |
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| YU Xi-yan,TIAN Hong-mei,FAN Ji-de,WANG Xiu-feng.Construction and study of a vector for antisense expression of acid invertase gene from melon[J].Journal of Fruit Science,2006,23(6):850-853. |
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| Authors: | YU Xi-yan TIAN Hong-mei FAN Ji-de WANG Xiu-feng |
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| Abstract: | Improvement of fruit quality is an important object in melon breeding. Application of antisense RNA strategy to reduce acid invertase activity provides a new feasible method for breeding melon cultivars with high sucrose accumulation. In a previous study we cloned a cDNA fragment of acid invertase from melon fruit into pMD18-T. For constructing antisense melon acid invertase gene vector (Anti-MAI1), the gene was digested with BamH Ⅰ and Hinc Ⅱ to release a fragment with 1038 bp coding region,which was then inserted between the cloning sites of BamH Ⅰ and Sma Ⅰ of pROK2. Anti-MAI1 was transferred into tobacco mediated by Agrobacterium tumefacions LBA4404. PCR and PCR-Southern blot were performed,and the results showed that MAI1 had been transformed into the genomic DNA of tobacco. |
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| Keywords: | Melon Acid invertase gene Antisense expression vector Tobacco Transformation |
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