Open AccessReverse transcriptase in archaebacteria
Author(s) -
BENMAHREZ Kamel,
SOROKINE Iréne,
NAKAYAMA Masachi,
KOHIYAMA Masamichi
Publication year - 1991
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1991.tb15689.x
Subject(s) - klenow fragment , dna polymerase , reverse transcriptase , dna polymerase i , primase , primer (cosmetics) , rnase h , microbiology and biotechnology , biochemistry , biology , polymerase , dna , oligonucleotide , enzyme , dna clamp , exonuclease , chemistry , polymerase chain reaction , organic chemistry , gene
A primase – reverse‐transcriptase of Halobacterium halobium was purified by column chromatography on DEAE‐cellulose, hydroxyapatite and carboxymethyl‐cellulose, followed by sedimentation on a glycerol gradient. The enzyme is a multifunctional enzyme containing reverse transcriptase, DNA polymerase and RNase H activities and does not require a preformed primer to initiate DNA synthesis. Using a single‐stranded DNA as template, this enzyme synthesizes oligonucleotides (8 – 12 bases) that can be used as primer by Escherichia coli DNA nucleotidyltransferase I (DNA polymerase I, Klenow fragment). Two polypeptides of 67 and 57 kDa were found after 14750‐fold purification of the enzyme.