       Document 0305
 DOCN  M9640305
 TI    Requirements for optimal expression of secreted and nonsecreted
       recombinant proteins in vaccinia virus systems.
 DT    9604
 AU    Pfleiderer M; Falkner FG; Dorner F; Immuno AG Biomedical Research
       Center, Orth/Donau, Austria.
 SO    Protein Expr Purif. 1995 Oct;6(5):559-69. Unique Identifier : AIDSLINE
       MED/96106929
 AB    Selection of an optimal promoter is necessary for efficient expression
       of foreign genes with vaccinia virus. Since a variety of powerful
       (homologous) vaccinia virus promoters and foreign (heterologous)
       promoter systems have been described for use in vaccinia, we have
       addressed the question of whether a general rule exists that allows the
       prediction of the optimal promoter/gene combination. We have compared
       the expression properties of four secreted proteins, the human blood
       clotting factor IX (FIX), the human blood glycoprotein Protein S
       (ProtS), the human von Willebrand factor (vWF), and the Hepatitis B
       virus (HBV) middle surface glycoprotein preS2, with proteins that were
       reported not to be secreted, the HBV large surface glycoprotein preS1
       and the murine leukemia virus (MuLV) BM-5 Eco gag protein. In addition,
       we have included in our study an internal control protein, the vaccinia
       virus p11 protein, to monitor possible side effects of the promoter
       system used. Genes encoding the foreign proteins were placed either
       under control of a synthetic vaccinia virus early/late promoter (selP)
       or under control of the bacteriophage T7 promoter (T7/emc system). The
       secreted proteins were more efficiently expressed when fused to the
       homologous promoter. Direct comparison of the two promoters indicated
       that the expression level ranged between 1.4 (ProtS) and 3.9 (FIX)-fold
       higher with the selP than with the T7 promoter. In contrast, the
       cell-associated HBV preS1 was more efficiently expressed under the T7
       promoter and the MuLV BM-5 Eco gag polypeptide was expressed equally
       well from both promoters. These data indicate that a careful prediction
       of optimal promoter/foreign gene combinations for the vaccinia virus
       expression system is possible. The choice of the optimal
       promoter/expression system is based on a simple classification scheme,
       discriminating secreted and nonsecreted proteins.
 DE    von Willebrand Factor/BIOSYNTHESIS/GENETICS  Animal  Bacteriophage T7
       Base Sequence  Cell Line  Comparative Study  Cytoplasm  Factor
       IX/BIOSYNTHESIS/GENETICS  Gene Expression  Gene Products,
       gag/BIOSYNTHESIS/GENETICS  Glycoproteins/BIOSYNTHESIS/GENETICS
       Haplorhini  Hepatitis B Virus/CHEMISTRY  Human  Leukemia Viruses,
       Murine/CHEMISTRY  Molecular Sequence Data  Promoter Regions (Genetics)
       Protein S/BIOSYNTHESIS/GENETICS  Recombinant Fusion
       Proteins/*BIOSYNTHESIS/CLASSIFICATION/GENETICS  Transcription, Genetic
       Vaccinia Virus/*GENETICS  Viral Envelope Proteins/BIOSYNTHESIS/GENETICS
       JOURNAL ARTICLE

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

