title: Cloning and high level nonfusion expression of recombinant human basic fibroblast growth factor in Escherichia coli.
abstract: OBJECTIVE
  To obtain high-level expression of nonfusion recombinant human basic fibroblast growth factor (rhbFGF).
  
  
  METHODS
  hbFGF cDNA was prepared from the total RNA of embryonic brain tissue. As a template, the obtained gene was used to clone nonfusion rhbFGF. New primers were employed to alter the translation initiation region (TIR) and reduce the G+C content through nucleotide change. Using pET-3C as vector, the cloned rhbFGF was expressed in BL21 (DE3).
  
  
  RESULTS
  rhbFGF was expressed in E coli up to 30 % of the total cellular protein. Cation exchange and heparin affinity chromatography were employed to purify the target protein from the supernatant of bacteria lysate. The bioactivity of the purified rhbFGF was identical with the standard bFGF.
  
  
  CONCLUSIONS
  Modification of TIR is an effective means to increase nonfusion expression rate of recombinant proteins, such as rhbFGF, in E coli.
authors.affiliation: Bioengineering Institute of Jinan University, Guangzhou 510632, China. ojds@jnu.edu.cn
authors.forename: Xiao-Jia
authors.initials: XJ
authors.lastname: Chen
authors.forename: Fen-Yong
authors.initials: FY
authors.lastname: Sun
authors.forename: Qiu-Ling
authors.initials: QL
authors.lastname: Xie
authors.forename: Mei-De
authors.initials: MD
authors.lastname: Liao
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhi-Ying
authors.initials: ZY
authors.lastname: Li
authors.forename: An
authors.initials: A
authors.lastname: Hong
authors.forename: Jian
authors.initials: J
authors.lastname: Lin
mesh_terms: D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005333:Fetus; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis* / Q000235:genetics; D005815:Genetic Code; D006801:Humans; D008969:Molecular Sequence Data; D011506:Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D011506:Proteins; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2