title: High-level production of active HIV-1 protease in Escherichia coli.
abstract: High levels of active HIV-1 protease (PR) were produced in Escherichia coli, amounting to 8-10% of total cell protein. High production levels were achieved by altering the following parameters: (1) codon preference of the coding region, (2) A+T-richness at the 5' end of the coding region, and (3) promoter. To circumvent the toxicity of HIV-1 PR in E. coli, the gene was expressed as a fusion protein with two different proteolytic autocleavage sequences. In both the cases, the fusion protein could be cleaved in vivo to give an active molecule with the native sequence at the N terminus.
authors: Monsanto Corporate Research, Monsanto Co., St. Louis, MO 63198.
authors: S H
authors: SH
authors: Rangwala
authors: R F
authors: RF
authors: Finn
authors: C E
authors: CE
authors: Smith
authors: S A
authors: SA
authors: Berberich
authors: W J
authors: WJ
authors: Salsgiver
authors: W C
authors: WC
authors: Stallings
authors: G I
authors: GI
authors: Glover
authors: P O
authors: PO
authors: Olins
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005813:Genes, Synthetic; D016333:HIV Protease / Q000096:biosynthesis* / Q000235:genetics; D015497:HIV-1 / Q000201:enzymology*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid
chemical_list: D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins; D016333:HIV Protease