Document 0049
 DOCN  M95B0049
 TI    Stringent chemical and thermal regulation of recombinant gene expression
       by vaccinia virus vectors in mammalian cells.
 DT    9511
 AU    Ward GA; Stover CK; Moss B; Fuerst TR; MedImmune, Inc., Gaithersburg, MD
       20878, USA.
 SO    Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):6773-7. Unique Identifier :
       AIDSLINE MED/95350151
 AB    We developed a stringently regulated expression system for mammalian
       cells that uses (i) the RNA polymerase, phi 10 promoter, and T phi
       transcriptional terminator of bacteriophage T7; (ii) the lac repressor,
       lac operator, rho-independent transcriptional terminators and the gpt
       gene of Escherichia coli; (iii) the RNA translational enhancer of
       encephalomyocarditis virus; and (iv) the genetic background of vaccinia
       virus. In cells infected with the recombinant vaccinia virus, reporter
       beta-galactosidase synthesis was not detected in the absence of inducer.
       An induction of at least 10,000- to 20,000-fold occurred upon addition
       of isopropyl beta-D-thiogalactopyranoside or by temperature elevation
       from 30 to 37 degrees C using a temperature-sensitive lac repressor.
       Regulated synthesis of the secreted and highly glycosylated human
       immunodeficiency virus 1 envelope protein gp120 was also demonstrated.
       Yields of both proteins were approximately 2 mg per 10(8) cells in 24
       hr. Plasmid transfer vectors for cloning and expression of complete or
       incomplete open reading frames in recombinant vaccinia viruses are
       described.
 DE    beta-Galactosidase/BIOSYNTHESIS/GENETICS  Animal  Base Sequence  Cells,
       Cultured  Cloning, Molecular/*METHODS  Enzyme Induction  *Gene
       Expression Regulation  Genetic Vectors/*GENETICS  Heat  HIV Envelope
       Protein gp120/BIOSYNTHESIS/GENETICS  Isopropyl
       Thiogalactoside/PHARMACOLOGY  Molecular Sequence Data  Recombinant
       Proteins/BIOSYNTHESIS  Regulatory Sequences, Nucleic Acid/GENETICS
       Support, U.S. Gov't, P.H.S.  Vaccinia Virus/*GENETICS  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).