Chinese Journal of Tissue Engineering Research ›› 2012, Vol. 16 ›› Issue (28): 5246-5250.doi: 10.3969/j.issn.2095-4344.2012.28.024

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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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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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