title: A set of beta-galactosidase gene fusion cassettes demonstrates usefulness in expressing HIV-1 genes in Escherichia coli.
abstract: Heterologous expression in Escherichia coli is often limited by yield and solubility of the foreign protein in the bacterial cytoplasm. In many cases, overexpression also results in growth inhibition. In order to produce retroviral proteins that are especially difficult to overexpress in E. coli, we designed a set of beta-galactosidase fusion cassettes. Fusions with beta-galactosidase increase significantly both yield and solubility of the foreign proteins, thus making purification much easier. These cassettes allow for N- or C-terminal fusions, and the retroviral proteins can be released from the fusion by automaturation in vivo for the HIV-1 protease or cleavage by thrombine for Tat. More generally, any synthetic sequence coding for a given cleavage site can be introduced 5' or 3' to the lacZ gene through a convenient set of unique restriction sites, making these fusion cassettes highly versatile.
authors.affiliation: Institut National des Sciences Appliquées, CNRS, Toulouse, France.
authors.forename: V
authors.initials: V
authors.lastname: Valverde
authors.forename: H
authors.initials: H
authors.lastname: Duplan
authors.forename: M
authors.initials: M
authors.lastname: Knibiehler
authors.forename: J M
authors.initials: JM
authors.lastname: Masson
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D015756:Genes, tat; D015497:HIV-1 / Q000235:genetics*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D012191:Retroviridae Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D001616:beta-Galactosidase / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D012191:Retroviridae Proteins; D001616:beta-Galactosidase