title: Translation initiation region plays an important role in the expression of human thrombopoietin in Escherichia coli.
abstract: A mutant human thrombopoietin (TPO) gene with a modified translation initiation region (TIR) sequence was created by site-specific mutagenesis based on the PCR technique. This mutant TPO gene encoded the same amino acid sequence as wild-type TPO gene. The wild-type TPO gene was expressed in E. coli with very low efficiency. The mutant TPO gene could reach an expression level of up to 10% of total cellular proteins in E. coli, which was much higher than the wild-type gene. The recombinant protein was mainly in the form of inclusion body which could acquire in vitro activities of human thrombopoietin after refolding.
authors.affiliation: Department of Biochemistry, Nanjing University, P. R. China.
authors.forename: X
authors.initials: X
authors.lastname: Jiang
authors.forename: S
authors.initials: S
authors.lastname: Li
authors.forename: A
authors.initials: A
authors.lastname: Zhou
authors.forename: F
authors.initials: F
authors.lastname: Li
authors.forename: X
authors.initials: X
authors.lastname: Xu
authors.forename: D
authors.initials: D
authors.lastname: Zhu
mesh_terms: D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013926:Thrombopoietin / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D013926:Thrombopoietin