High‐level expression of enzymatically active bovine leukemia virus proteinase in E. coli

FEBS Letters

Hořejši, Magda¹  Konvalinka, Jan²  Sedláčeka, Juraj¹  Štrop, Petr²  Fábry, Milan¹  Hrušková-Heidingsfeldová, Olga²  Andreánsky, Martin¹  Bláha, Ivo²  Ječmen, Petr¹ 

[1] Czechoslovak Academy of Sciences, Institute of Molecular Genetics, 166 37 Prague 6, Czechoslovakia;Institute of Organic Chemistry and Biochemistry, 166 37 Prague 6, Czechoslovakia

关键词: Retroviral proteinase;    Polyprotein precursor processing;    Recombinant product accumulation;    Synthetic peptide substrate;    Bovine leukemia virus;    E. coli;   

DOI  :  10.1016/0014-5793(91)80032-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

An E. coli plasmid expressing efficiently an artificial precursor of bovine leukemia virus (BLV) proteinase under transcriptional control of the phage T7 promoter was constructed. The expression product accumulates in the induced E. coli cells in the form of insoluble cytoplasmic inclusions. Solubilization of the inclusions and a refolding step yield almost pure and completely self-processed proteinase. Purification to homogeneity was achieved by ion-exchange chromatography and reverse-phase HPLC. On a preparative scale, a high yield of enzymatically active proteinase was obtained. An initial study using a series of synthetic peptide substrates shows a distinct substrate specificity of BLV proteinase.

【 授权许可】

Unknown