中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (28): 5246-5250.doi: 10.3969/j.issn.2095-4344.2012.28.024

• 组织构建基础实验 basic experiments in tissue construction • 上一篇    下一篇

降钙素基因相关肽表达载体的构建

刘成龙，吕 海，邱素均，段 扬   

  1. 南方医科大学珠江医院骨科 ，广东省广州市 510282
  • 收稿日期:2012-02-21 修回日期:2012-05-14 出版日期:2012-07-08 发布日期:2012-07-08
  • 作者简介:刘成龙☆，男，1973年生，山西省太原市人，汉族，2003年解放军第一军医大学毕业，博士，副主任医师，主要从事脊柱退行性病变及脊柱脊髓损伤的研究。 liucw878@163.com

Construction of an expression vector of calcitonin gene-related peptide gene

Liu Cheng-long, Lü Hai, Qiu Su-jun, Duan Yang   

  1. Department of Othopedics, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China
  • Received:2012-02-21 Revised:2012-05-14 Online:2012-07-08 Published:2012-07-08
  • About author:Liu Cheng-long☆, Doctor, Associated physician, Department of Othopedics, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, Guangdong Province, China liucw878@163.com

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392

被引次数

2

摘要：

背景：为研究降钙素基因相关肽基因治疗在骨质疏松中的应用，首先要建立高效的降钙素基因相关肽基因载体。
目的：构建降钙素基因相关肽真核表达载体pBaBb-puro-CGRP。
方法：设计引物并通过PCR扩增出降钙素基因相关肽，酶切后用T4 DNA连接酶将其与pBaBb-puro进行连接，经过转化、阳性克隆筛选后，进行酶切和测序鉴定。
结果与结论：经PCR扩增获得了含430 bp的降钙素基因相关肽基因编码序列，经过酶切、连接、转化后，挑选10个菌落进行PCR检测，筛查到8个菌落含有重组质粒，进一步行酶切和测序鉴定，结果与理论预期完全相符。提示实验成功构建了pBaBb-puro-CGRP真核表达载体。

关键词: 降钙素基因相关肽, 载体, 构建, 骨质疏松, 组织构建

Abstract:

BACKGROUND: Efficient calcitonin gene-related peptide (CGRP) gene vector should be established first to study the application of CGRP gene therapy in osteoporosis.
OBJECTIVE: To construct an expression vector of pBaBb-puro-CGRP.
METHODS: Primers were designed. Then, CGRP was obtained by PCR amplification and it was connected to pBaBb-puro after enzyme digestion. After transformation and positive clone screening, enzyme digestion and sequence identification were performed.
RESULTS AND CONLUSION: 430 bp coding sequence of CGRP gene was obtained by PCR amplification. After enzyme digestion, connection and transformation, the clones were obtained. Next, 10 clones were picked up and examined by PCR, and eight clones were found the recombinant plasmid. The result was in accord with expectation through further enzyme digestion and sequencing. These findings suggest that expression vector of pBaBb-puro-CGRP was successfully constructed.

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