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Translation initiation region plays an important role in the expression of human thrombopoietin in Escherichia coli
Author(s) -
Jiang Xueyuan,
Li Suqin,
Zhou Aiwu,
Li Faqing,
Xu Xianxiu,
Zhu Dexu
Publication year - 1996
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549600201282
Subject(s) - thrombopoietin , mutant , gene , biology , microbiology and biotechnology , recombinant dna , mutagenesis , escherichia coli , gene expression , translation (biology) , eukaryotic translation , wild type , mutation , site directed mutagenesis , genetics , messenger rna , stem cell , haematopoiesis
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.