Open AccessExpression in Escherichia coli of a fusion protein product containing a region of the adenovirus DNA polymerase.
Author(s) -
David Rekosh,
Jeff Lindenbaum,
James M. Brewster,
Lawrence M. Mertz,
Jerard Hurwitz,
Laura Prestine
Publication year - 1985
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.82.8.2354
Subject(s) - microbiology and biotechnology , polymerase , fusion protein , dna polymerase , biology , dna polymerase i , escherichia coli , dna polymerase ii , adenovirus genome , cloning (programming) , adenoviridae , dna , plasmid , polymerase chain reaction , virology , recombinant dna , gene , genetics , reverse transcriptase , computer science , programming language
The bulk of an open reading frame extending from map coordinates 23.3 to 14.2 in region E2b of the adenoviral genome has been cloned and expressed from a chimeric plasmid in Escherichia coli. The cloning strategy used created a fusion protein of 124,000 daltons, which contained greater than 98% adenovirus-encoded sequences. Antiserum raised against this protein reacted with the authentic 140,000-dalton adenovirus DNA polymerase. Another serum raised against a synthetic hexapeptide whose sequence corresponded to the predicted carboxyl terminus of adenovirus-encoded DNA polymerase also reacted with the fusion protein and authentic adenovirus DNA polymerase. These results demonstrate that the cloned region of DNA encodes the adenovirus DNA polymerase.