rdf-syntax-grammar
mods:
  schema location: http://www.loc.gov/mods/v3 http://www.loc.gov/standards/mods/v3/mods-3-2.xsd
  title info:
    title: Purification and functional characterization of several essential DNA replication proteins from Autographa californica polyhedrosis virus
  name:
    name part: Hang, Xin
    name part: 1969-
    role:
      role term: creator
  type of resource: text
  genre: bibliography
  origin info:
    place:
      place term: xx
    date issued: 1996
    issuance: monographic
  language:
    language term: eng
  physical description:
    form: print
    extent: xi, 128 leaves : illustrations ; 28 cm.
  abstract: The Autographa californica polyhedrosis virus (ACMNPV) replicates in the nuclei of infected cells and encodes several proteins required for viral DNA replication. Transient replication assay of plasmids containing a viral origin have shown that at least six viral genes (dna pol, helicase, lef-1, lef-2, lef-3, and iel) are essential while products of the ie2, pe38, p35, and lej7 genes stimulate the level of DNA replication. To functionally characterize viral replication proteins, we purified single-stranded binding protein (SSB) from AcMNPV-infected insect cells using affinity chromatography. Control experiments using uninfected cell extracts suggested that the SSB was encoded by the virus. SSB bound to single-stranded DNA in solution, and competition binding experiments showed that SSB binds to ssDNA preferentially and that it binds to ssDNA in a non-sequence-specific manner. To determine whether SSB was encoded by the ACMNPV lef-3 gene, we cloned the lef-3 open reading frame under the control of the bacteriophage T7 promoter. Immunochemical analysis indicated that LEF-3 produced in bacteria or in rabbit reticulate lysates specifically reacted with monoclonal antibodies raised against purified SSB and biochemical analysis indicated purified LEF-3 from bacteria overexpression systems bound to single-stranded DNA in solution. In addition, temporal expression of LEF-3/SSB during virus infection was examined by inimunoblot analysis. The viral DNA polymerase was purified to near homogeneity from infected cell nuclear extracts using conventional chromatographic methods. Purified viral DNA polymerase had intrinsic 3'-5' exonuclease activity and was sensitive to several metabolic drugs. Template challenge experiments using singly-primed ssDNA templates indicated that the baculovirus DNA polymerase was highly processive. As initial steps to understand the functions of AcMNPV LEF-I and LEF-2 proteins, the genes were cloned into bacterial expression plasmids and purified to ne…
  note: by Xin Hang.
  note: Vita.
  note: "Major Subject: Biochemistry".
  note: Issued also on microfiche from University Microfilms Inc.
  note: Includes bibliographical references: pages 116-126.
  note: Ph. D. Texas A&M University 1996.
  subject:
    topic: Major biochemistry
  identifier: 97-01642 UMI
  identifier: http://proxy.library.tamu.edu/login
  related item:
    location:
      url: http://proxy.library.tamu.edu/login
  record info:
    record content source: TXA
    record creation date: 970501
    record change date: 20190325093255.0
    record identifier: in00001345678