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| 家兔C型利钠肽基因的克隆及鉴定 | |
| 引用本文： | 张剑,龚昆梅,肖乐,王昆华,欧阳一鸣,李临海,朱宇,龙亚新. 家兔C型利钠肽基因的克隆及鉴定[J]. 山东医药, 2011, 51(46): 24-25 |
| 作者姓名： | 张剑 龚昆梅 肖乐 王昆华 欧阳一鸣 李临海 朱宇 龙亚新 |
| 作者单位： | 昆明医学院附属昆华医院,昆明,650034 |
| 基金项目： | 国家自然科学基金资助项目(30860276); 云南省科技厅科研基金资助项目(2007C248M);云南省科技厅—昆明医学院应用基础专项基金资助项目(2008GD042) |
| 摘 要： | 目的克隆家兔C型利钠肽基因，为其真核表达载体的构建奠定基础。方法通过RT-PCR从家兔腹主动脉组织中克隆C型利钠肽基因全长编码区，将该DNA片段亚克隆至克隆载体pMD18-T中，用Pst Ⅰ单酶切筛选出负向插入片段，对重组质粒进行鉴定。结果PCR、酶切及测序鉴定证实，家兔C型利钠肽基因全长编码区被准确插入到载体pMD18-T的TA克隆位点中。结论成功克隆家兔c型利钠肽基因，为其真核表达载体的构建及开展糖尿病足C型利钠肽基因治疗奠定了基础。 |
| 关 键 词： | C型利钠肽 基因克隆 糖尿病足 兔 |
Cloning and sequencing of rabbit C-type natriuretic peptide gene |
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| ZHANG Jian , GONG Kun-mei , XIAO Le , WANG Kun-hua , OUYANG Yi-min , LI Ling-hai , ZHU Yu , LONG Ya-xin. Cloning and sequencing of rabbit C-type natriuretic peptide gene[J]. Shandong Medical Journal, 2011, 51(46): 24-25 | |
| Authors: | ZHANG Jian GONG Kun-mei XIAO Le WANG Kun-hua OUYANG Yi-min LI Ling-hai ZHU Yu LONG Ya-xin |
| Affiliation: | ZHANG Jian,GONG Kun-mei,XIAO Le,WANG Kun-hua,OUYANG Yi-min,LI Ling-hai,ZHU Yu,LONG Ya-xin (Kunhua Affiliated Hospital of Kunming Medical College,Kunming 650032,P.R.China) |
| Abstract: | Objective To clone and analyze the sequence of rabbit C-type natriuretic peptide gene and lay the foundation of construct the recombinant eukaryotic vector. Methods The full coding domain sequence of rabbit C-type natriuretic peptide gene was cloned from rabbit abdominal aorta with RT-PCR and was sub-cloned into pMD 18-T vector. After screening of negative pMD 18-T/CNP with restriction enzyme Pst Ⅰ degestion. Then, the reconstructed plasmid was identified with enzyme digestion, PCR and sequencing. Results It was demonstrated that the full coding domain sequence of rabbit C-type natriuretic peptide gene was accurately cloned into the sites of pMD 18-T vector. Conclusions Rabbit C-type natriuretic peptide gene is donen successfully, it lay the groundwork for constructing the recombinant eukaryotic vector and gene therapy for diabetic foot patients with C-type natriuretic peptide. |
| Keywords: | C-type natriuretic peptide gene cloning diabetic foot rabbit |
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