| 不结球白菜L-半乳糖酸-1,4-内酯脱氢酶基因(GLDH)cDNA片段克隆与RNAi表达载体构建 |
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| 引用本文： | 高红亮,李英,宋玉萍,马成英,侯喜林.不结球白菜L-半乳糖酸-1,4-内酯脱氢酶基因(GLDH)cDNA片段克隆与RNAi表达载体构建[J].南京农业大学学报,2008,31(4). |
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| 作者姓名： | 高红亮 李英 宋玉萍 马成英 侯喜林 |
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| 作者单位： | 南京农业大学园艺学院/作物遗传与种质创新国家重点实验室,江苏,南京,210095 |
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| 摘 要： | 根据植物中hpRNA(hairpin RNA)原理,选择GLDH基因cDNA序列同源性较低区域设计1对携带双酶切位点的特异引物,以不结球白菜‘苏州青’叶片为材料提取总RNA,利用RT-PCR技术克隆了GLDH基因片段。构建中间克隆载体,经过3次亚克隆将测序正确的GLDH基因片段分别以正向和反向两种形式插入到间隔基因YYT的两端,经鉴定已插入到植物表达载体中,成功地构建了干扰GLDH基因表达的RNAi植物双元表达载体pRNAi-GLDH,并将表达载体转化到农杆菌菌株LBA4404中,为深入研究GLDH基因功能提供有效的工具。
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| 关 键 词： | 不结球白菜 GLDH基因 RNAi植物双元表达载体 |
Cloning of GLDH gene cDNA fragment from non-heading Chinese cabbage and construction of its corresponding RNAi binary vector |
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| GAO Hong-liang,LI Ying,SONG Yu-ping,MA Cheng-ying,HOU Xi-lin.Cloning of GLDH gene cDNA fragment from non-heading Chinese cabbage and construction of its corresponding RNAi binary vector[J].Journal of Nanjing Agricultural University,2008,31(4). |
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| Authors: | GAO Hong-liang LI Ying SONG Yu-ping MA Cheng-ying HOU Xi-lin |
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| Abstract: | According to the principle of hpRNA(hairpin RNA)in the plant,a cDNA fragment encoding L-galactono-1,4-lactone dehydrogenase(GLDH)was cloned from non-heading Chinese cabbage by the method of RT-PCR on the basis of the specific primers with double enzyme sites.GLDH fragment was ligated with YYT interval region at forward and reverse orientations through three sub-cloning via mid-clone vector.The fusion gene was inserted into plant vector to construct the recombinant plasmid pRNAi-GLDH,which was then transformed into Agrobacterium LBA4404.This will provide an effective tool for the further study of GLDH gene function. |
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| Keywords: | non-heading Chinese cabbage(Brassica campestris ssp chinensis Makino) GLDH RNAi binary expression vector |
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