Construct the recombinant plasmid of recombinant human basic fibroblast growth factor
10.3760/cma.j.issn.1008-1372.2012.05.005
- VernacularTitle:人碱性成纤维生长因子重组质粒的构建
- Author:
Lihong FAN
;
Xueqing CHEN
;
Kequan CHEN
;
Xiaoqiang YANG
;
Zhenshu ZHANG
- Publication Type:Journal Article
- Keywords:
Recombinant proteins/genetics;
Fibroblast growth factor 2/genetics;
Plasmids;
Genetic vectors
- From:
Journal of Chinese Physician
2012;14(5):593-595,599
- CountryChina
- Language:Chinese
-
Abstract:
Objective To obtain the sequence of recombinant human basic fibroblast growth factor (rhbFGF) with endonuclease sites of BamHI and Pst Ⅰ by PCR based gene assembly and construct the recombining vector of rhbFGF gene by TA cloning technique.Methods The rhbFGF gene sequence which was designed for lactococcus with endonuclease sites of BamHI and Pst Ⅰ was divided into 22 oligonucleotides by DNASTAR 6.0 (bFGFl-bFGF22).The 22 oligonucleotides were spliced by PCR based gene assembly to get the rhbFGF eDNA with endonuclease sites of BamHI and Pst Ⅰ.The PCR product was inserted into the PMD18-T VECTOR.The recombining vector were converted to the competent E.coli TOP10.The clones generated from LAB were analyzed by miniprep isolation from LAB host.They were identified by the restriction enzyme cuuing and sequencing.Results The rhbFGFcDNA synthesized by PCR based gene assembly with endonuclease sites of BamHI and Pst Ⅰ was verified by 2％ agarose electrophoresis.The recombining vector of rhbFGF gene by TA cloning technique was identified by enzyme digestion and gene sequencing.Conclusions The TA cloning vector of recombining hbFGF with endonuclease sites of BamHI and Pst Ⅰ was constructed successfully.
