| 摘要: |
| ［摘要］ 目的 构建CCL11、CCL26基因3′非翻译区（3′UTR）真核表达载体。方法 多聚酶链式反应（PCR）扩增获得CCL11、CCL26基因3′UTR目的片段，双酶切目的基因片段及pMIR REPORT真核表达质粒后，将目的基因片段插入pMIR REPORT质粒，再将重组质粒转化感受态DH5α菌株，筛选阳性克隆行双酶切鉴定及测序分析。结果 成功构建CCL11、CCL26基因3′UTR真核表达载体。结论 CCL11、CCL26基因3′UTR真核表达载体成功建立，为进一步研究CCL11、CCL26基因与MicroRNAs的关系提供实验基础。 |
| 关键词: CCL11基因 CCL26基因 3′非翻译区 真核表达载体 pMIR REPOR |
| DOI：10.3969/j.issn.1674-3806.2014.01.02 |
| 分类号:Q 782 |
| 基金项目:广西自然科学基金资助项目（编号:2011GXNSFC018019） |
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| Construction of eukaryotic expression vectors of CCL11 gene 3′UTR and CCL26 gene 3′UTR and it′s significance |
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WEN Zhi-hong, DAI Yan, HE Shuang
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Department of Pediatrics, the People′s Hospital of Guangxi Zhuang Autonomous Region, Nanning 530021, China
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| Abstract: |
| ［Abstract］ Objective To construct the eukaryotic expression vectors of 3′untranslated regions（3′UTR） of CCL11 gene and CCL26 gene.Methods Target fragments in 3′UTR of CCL11 gene and CCL26 gene were amplified from genomic DNA by Polymerase chain reaction(PCR).The target fragments and pMIR REPORT vectors were double cut by restriction enzyme,and then the fragments were inserted into the pMIR REPORT plasmid to construct recombinant plasmid.The recombinant plasmid of pMIR REPORT-CCL11 3′UTR and pMIR REPORT- CCL26 3′UTR were transformed DH5α competent cells,respectively.Results The 3′UTR sequence of CCL11 gene and CCL26 gene were successfully cloned into the pMIR REPORT vector,which verified by restrictive enzyme digestion and DNA sequencing.Conclusion The recombinant plasmid of pMIR REPOR-CCL11 3′UTR and pMIR REPORT- CCL26 3′UTR were successfully constructed,which lay a foundation of the further study of the relation between CCL11 gene,CCL26 gene and microRNAs. |
| Key words: CCL11 gene CCL26 gene 3′UTR Eukaryotic expression vector pMIR REPOR |
