| 微绿球藻DNA质粒文库的构建 |
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| 引用本文： | 赵大显,周志刚. 微绿球藻DNA质粒文库的构建[J]. 海洋科学, 2006, 30(5): 87-91 |
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| 作者姓名： | 赵大显 周志刚 |
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| 作者单位： | 上海水产大学,农业部水产种质资源与养殖生态重点开放实验室,生命科学与技术学院,上海,200090;上海水产大学,农业部水产种质资源与养殖生态重点开放实验室,生命科学与技术学院,上海,200090 |
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| 基金项目： | 国家转基因植物研究与产业化开发专项(JY03-B-20)，上海市属高校自然科学研究资助项目(01H04) |
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| 摘 要： | 微绿球藻(Nannochloropsis oculata)DNA被提取纯化并经超声波处理后,将所得大小在1.6~3 kb之间的DNA片段用T4 DNA聚合酶补平,再与SmaI酶切消化并经去磷酸化处理的质粒载体pUC18连接,转化至DH10B大肠杆菌(Escherichia coli)的感受态细胞中。建立的DNA质粒文库容量含2×104个克隆,其中重组子占90%。随机挑选白色菌落并培养,抽提的质粒经XbaI和SacI双酶切鉴定,显示重组的质粒中均含有大小不等的DNA插入片段。
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| 关 键 词： | 微绿球藻(Nannochloropsis oculata) 超声处理 质粒载体pUC18 DNA文库 |
| 文章编号： | 1000-3096（2006）05-0087-05 |
| 收稿时间： | 2004-06-30 |
| 修稿时间： | 2005-02-20 |
DNA library of Nannochloropsis oculata constructed with plasmid vector pUC18 |
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| ZHAO Da-xian,ZHOU Zhi-gang. DNA library of Nannochloropsis oculata constructed with plasmid vector pUC18[J]. Marine Sciences, 2006, 30(5): 87-91 |
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| Authors: | ZHAO Da-xian ZHOU Zhi-gang |
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| Abstract: | The genomic DNA isolated and purified from Nannochloropsis oculata was sheared after a treatment with an ultrasonic processor. The treated DNA was blunt-ended with T4 DNA polymerase and then was collected from 1.6 kb to 3 kb in size. The target blunt-ended DNA was cloned into plasmid vector pUC18 which was previously digested with Sma I and dephosphorylated with calf alkaline phosphatase. The recombinant plasmid was transformed into Escherichia coli DH10B competent cells and the genomic library was constructed. There were about 2×104 clones in the genomic library of N. oculata, and the percentage of recombinants was about 90%. It was confirmed that all randomly selected plasmids of white colonies contained target DNA inserts from 1.6 kb to 3 kb in size after restriction mapping with Xba I and Sac I endonucleases. |
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| Keywords: | Nannochloropsis oculata ultrasonication plasmid vector pUC18 genomic library |
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