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| 胡萝卜抗冻蛋白基因克隆及植物表达载体构建 | |
| 引用本文： | 尹明安,崔鸿文,樊代明.胡萝卜抗冻蛋白基因克隆及植物表达载体构建[J].西北农林科技大学学报(社会科学版),2001,29(1):6-10. |
| 作者姓名： | 尹明安 崔鸿文 樊代明 |
| 作者单位： | 1. 西北农林科技大学 园艺学院, 2. 第四军医大学 西京医院, 3. 西北农林科技大学 生命科学学院, |
| 基金项目： | 陕西省科技攻关项目(91K12-G19(11)) |
| 摘 要： | 以胡萝卜品种 Autumn King的幼苗为材料 ,用 CTAB法提取其基因组 DNA,以 PCR ( Polym eraseChain Reaction)的方法在体外扩增出胡萝卜抗冻蛋白基因 ( afp) ,以 p UCm - T Vector为载体构建成胡萝卜 afp的克隆载体 p TAF,用 Eco R 消化重组质粒 p TAF使其线性化 ,再用 DNA聚合酶 Klenow大片段补平末端 ,然后用Xba 消化 ,获得一末端粘 ,一末端平的目的片段 ( afp)。植物表达载体 p BI12 1用 X ba 和 Sma 双酶切 ,获得一末端粘 ,一末端平的线性质粒。将目的片段与线性质粒在 T4DNA连接酶的作用下进行定向连接 ,构建成胡萝卜 afp的植物表达载体 p BAF |
| 关 键 词： | 胡萝卜 抗冻蛋白基因 分子生物学 |
| 文章编号： | 1000-2782(2001)01-0006-05 |
| 收稿时间： | 2000/10/31 0:00:00 |
| 修稿时间： | 2000年10月31 |
Cloning of a carrot gene encoding antifreeze protein and construction of its plant expression vector |
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| JIA Zhi kuan,DAI Jun.Cloning of a carrot gene encoding antifreeze protein and construction of its plant expression vector[J].Journal of Northwest Sci-Tech Univ of Agr and,2001,29(1):6-10. | |
| Authors: | JIA Zhi kuan DAI Jun |
| Institution: | 3 (1 College of Horticulture,Northwest Science and Technology University of Agriculture and Forestry, Yangling,Shaanxi 712100,China;2 Xijing Hospital,the Fourth University of Military Medicine,Xi |
| Abstract: | Genomic DNA in the seedlings of carrot cultivar Autumn King was extracted with CTAB method.The carrot antifreeze protein gene (afp) was amplified by PCR(Polymerase Chain Reaction).Cloning vector pTAF of carrot afp was constru cted with pUCm-T Vector.PTAF was digested with EcoRⅠ and became linear.Its ends were filled with DNA Polymerase Ⅰ Klenow fragment.Then it was digested wi th XbaⅠ and a designed fragment (afp) with a cohesive end and a blunt end was released.Plant expression vector pBI121 was digested with XbaⅠ and SmaⅠand a linear plasmid with a cohesive end and a blunt end was obtained.The linear plasmid and the designed fragment (afp) were directively ligated with T4 DNA ligase,and the plant expression vector of carrot afp was constructed. |
| Keywords: | carrot antifreeze protein gene molecular biology |
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