| PCR方法在水蛭素基因克隆方面的新应用 |
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| 引用本文： | 翁梁,刘丹,姚为民,盛君.PCR方法在水蛭素基因克隆方面的新应用[J].中国生化药物杂志,2002,23(2):62-63. |
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| 作者姓名： | 翁梁 刘丹 姚为民 盛君 |
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| 作者单位： | 1. 长春中医学院附属医院新药研究中心,吉林长春,130021
2. 长春生物制品研究所,吉林长春130062 |
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| 摘 要： | 目的应用PCR方法获得水蛭素基因。方法在传统PCR方法的基础上进行了改进 ,根据已知天然水蛭素基因序列 ,设计了一对 12 3nt且 3′端具有 15nt互补序列的单链DNA ,通过低温复性使其互补结合后 ,进行延伸反应 ,得到微量双链水蛭素基因。再以该基因作为模板设计一对引物 ,进行常规PCR扩增反应。结果获得水蛭素基因。结论此法对于获得象水蛭素基因这类序列已知、片段不太长 ,且不易得到的基因提供了一种可行方法
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| 关 键 词： | 水蛭素 基因克隆 PCR |
| 文章编号： | 1005-1678(2002)02-0062-02 |
| 修稿时间： | 2001年3月19日 |
The new application of PCR method in cloning of hirudin gene |
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| WENG Liang ,LIU Dan ,YAO Wei min ,SHENG Jun.The new application of PCR method in cloning of hirudin gene[J].Chinese Journal of Biochemical Pharmaceutics,2002,23(2):62-63. |
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| Authors: | WENG Liang LIU Dan YAO Wei min SHENG Jun |
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| Institution: | WENG Liang 1,LIU Dan 2,YAO Wei min 2,SHENG Jun 2 |
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| Abstract: | PurposeThe aim is to obtain recombinant hirudin gene using PCR method.MethodsConstructing two single chain DNA of 123nt,there is 15nt complementary sequence in 3′ end, performing extending reaction after complementary linking by renaturation at low temperature. The minimum double chain of hirudin gene was obtained. In the end, two primers on the template of hirudin gene were constructed by usual PCR.ResultsA large quantity of hirudin gene was established.ConclusionSince some known sequence such as hirudin gene, segment was short and obtaining whole gene segment was difficult, a more feasible method was established. |
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| Keywords: | hirudin gene clone PCR |
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