| 基于Xcm I酶切的pUC19-T载体的构建 |
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| 引用本文： | 孙程龙,李业伟,王颖,宫婷,扈荣良. 基于Xcm I酶切的pUC19-T载体的构建[J]. 安徽农业科学, 2011, 39(17): 10182-10184 |
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| 作者姓名： | 孙程龙 李业伟 王颖 宫婷 扈荣良 |
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| 作者单位： | 吉林大学畜牧兽医学院,吉林,长春,130062;军事医学科学院军事兽医研究所,吉林,长春,130122;军事医学科学院军事兽医研究所,吉林,长春,130122;军事医学科学院军事兽医研究所,吉林,长春,130122;吉林农业大学动物科技学院,吉林,长春,130122 |
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| 摘 要： | [目的]构建以pUC19质粒为基础可利用Xcm I内切酶制备的T栽体。[方法]化学合成2条含有双XcmI酶切位点的互补寡聚核苷酸链，经过变性、复性后克隆入pUCl9质粒的HindIII和BamHI位点之间，通过XcmI酶切后得到一个线性化的带有3’末端突出一个T碱基的T栽体。[结果]经TA克隆验证，制备的pUC19-HB-T载体对PCR产物的克隆率达到95％以上。[结论]构建的pUC19-HB-T载体可用于PCR产物的克隆、测序及后续分子生物学操作。
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| 关 键 词： | T载体 Xcm I限制性内切酶 TA克隆 |
Construction of A pUC19-T Vector Based on Xcm I |
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| SUN Cheng-long et al. Construction of A pUC19-T Vector Based on Xcm I[J]. Journal of Anhui Agricultural Sciences, 2011, 39(17): 10182-10184 |
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| Authors: | SUN Cheng-long et al |
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| Affiliation: | SUN Cheng-long et al (College of Animal Science and Veterinary Medicine,Jilin University,Changchun,Jilin 130062) |
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| Abstract: | [Objective] The paper aimed to construct a new T vector based on plasmid pUC19 digested with Xcm I. [Method] Two complementary oligonucleotide chains containing two Xcm I sites were synthesized. After denaturation and renaturation, the adaptor was cloned into plasmid pUC19 between the Hind III and BamH I sites. The new plasmid, pUC19-HB-T vector, was digested with Xcm I to derive a T-vector with 3′ end overhanging a T base. [Result] The constructed pUC19-HB-T vector was efficient in cloning PCR products, with an efficiency of 95% at least. [Conclusion] A new Xcm I-based pUC19-HB-T vector was constructed, which could be applied to cloning of PCR products and other microbiology operations. |
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| Keywords: | T vector Xcm I TA cloning |
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