HIGH-LEVEL EXPRESSION OF ENZYMATICALLY ACTIVE BOVINE LEUKEMIA-VIRUS PROTEINASE IN ESCHERICHIA-COLI

被引：16

作者：

ANDREANSKY, M ^{[1
]}

HRUSKOVAHEIDINGSFELDOVA, O ^{[1
]}

SEDLACEK, J ^{[1
]}

KONVALINKA, J ^{[1
]}

BLAHA, I ^{[1
]}

JECMEN, P ^{[1
]}

HOREJSI, M ^{[1
]}

STROP, P ^{[1
]}

FABRY, M ^{[1
]}

机构：

[1] CZECHOSLOVAK ACAD SCI, INST ORGAN CHEM & BIOCHEM, CS-16637 PRAGUE 6, CZECHOSLOVAKIA

来源：

FEBS LETTERS | 1991年 / 287卷 / 1-2期

关键词：

RETROVIRAL PROTEINASE; POLYPROTEIN PRECURSOR PROCESSING; RECOMBINANT PRODUCT ACCUMULATION; SYNTHETIC PEPTIDE SUBSTRATE; BOVINE LEUKEMIA VIRUS; ESCHERICHIA-COLI;

D O I：

10.1016/0014-5793(91)80032-X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

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 a 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.

引用

页码：129 / 132

页数：4

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