Premium
The terminal protein of the linear DNA plasmid pGKL2 shares an N‐terminal domain of the plasmid‐encoded DNA polymerase
Author(s) -
Takeda Masaharu,
Hiraishi Hiroyuki,
Takesako Toshikazu,
Tanase Sumio,
Gunge Norio
Publication year - 1996
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(19960315)12:3<241::aid-yea907>3.0.co;2-j
Subject(s) - biology , plasmid , kluyveromyces lactis , microbiology and biotechnology , dna , kluyveromyces , biochemistry , yeast , saccharomyces cerevisiae
The 36K protein attached at the 5′ end of the linear DNA plasmid pGKL2 from the yeast Kluyveromyces lactis was first purified and characterized. The terminal protein was purified from cells (1 kg wet weight) by ammonium sulphate precipitation and two rounds of centrifugation to equilibrium in CsCl gradients. The pGKL2 was present only in the post‐microsomal supernatant. Approximately 10 mg of the purified pGKL2 was recovered and digested with DNase I. The terminal protein (final ca. 0·8 mg) was homogeneous by electrophoresis and we determined the N‐terminal amino acid sequence up to ten residues, showing that it existed in the cryptic N‐terminal domain of pGKL2‐ORF2 (DNA polymerase) sequence.