Advanced

Molecular cloning and characterization of rat liver catechol-O-methyltransferase

Salminen, Marjo; Lundström, Kenneth; Tilgmann, Carola LU ; Savolainen, Raija; Kalkkinen, Nisse and Ulmanen, Ismo (1990) In Gene 93(2). p.241-247
Abstract

The coding sequence of rat liver catechol-O-methyl-transferase (COMT; EC 2.1.1.6) was determined from rat cDNA and genomic libraries were screened with DNA probes and specific antiserum. The open reading frame consisted of 663 nucleotides coding for a 221-amino acid (aa) polypeptide with a deduced Mr of 24747. No obvious hydrophobic signal sequence, membrane-spanning domains, or potential N-glycosylation sites were found in this sequence. The identity of the clone and the accuracy of the sequence was verified by direct aa sequencing of the tryptic peptides derived from the purified rat liver enzyme. Primer extension analysis showed that the transcription start point of the rat liver COMT mRNA was 450 bp upstream from the... (More)

The coding sequence of rat liver catechol-O-methyl-transferase (COMT; EC 2.1.1.6) was determined from rat cDNA and genomic libraries were screened with DNA probes and specific antiserum. The open reading frame consisted of 663 nucleotides coding for a 221-amino acid (aa) polypeptide with a deduced Mr of 24747. No obvious hydrophobic signal sequence, membrane-spanning domains, or potential N-glycosylation sites were found in this sequence. The identity of the clone and the accuracy of the sequence was verified by direct aa sequencing of the tryptic peptides derived from the purified rat liver enzyme. Primer extension analysis showed that the transcription start point of the rat liver COMT mRNA was 450 bp upstream from the translation start codon. A putative polyadenylation signal (ATTAAA) was found in the 3′-noncoding region. The predicted size of the COMT transcript was 1.8-2.0 kb, which could be confirmed from Northern hybridization analyses of the isolated rat liver mRNA. One polypeptide of 25 kDa, could be immunoprecipitated with anti-COMT antibody from in vitro translation of rat liver mRNA. Employing the DNA blot analysis only one COMT-encoding gene was found in the rat genome. © 1990.

(Less)
Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
keywords
genomic library, Methylating enzyme, nucleotide sequencing, recombinant DNA, RNA analysis, λ gt11 expression screening
in
Gene
volume
93
issue
2
pages
7 pages
publisher
Elsevier
external identifiers
  • Scopus:0025187695
ISSN
0378-1119
DOI
10.1016/0378-1119(90)90231-F
language
English
LU publication?
no
id
25536200-212b-4b69-b338-db4c9809a23c
date added to LUP
2016-04-11 13:26:13
date last changed
2016-07-27 09:35:17
@misc{25536200-212b-4b69-b338-db4c9809a23c,
  abstract     = {<p>The coding sequence of rat liver catechol-O-methyl-transferase (COMT; EC 2.1.1.6) was determined from rat cDNA and genomic libraries were screened with DNA probes and specific antiserum. The open reading frame consisted of 663 nucleotides coding for a 221-amino acid (aa) polypeptide with a deduced M<sub>r</sub> of 24747. No obvious hydrophobic signal sequence, membrane-spanning domains, or potential N-glycosylation sites were found in this sequence. The identity of the clone and the accuracy of the sequence was verified by direct aa sequencing of the tryptic peptides derived from the purified rat liver enzyme. Primer extension analysis showed that the transcription start point of the rat liver COMT mRNA was 450 bp upstream from the translation start codon. A putative polyadenylation signal (ATTAAA) was found in the 3′-noncoding region. The predicted size of the COMT transcript was 1.8-2.0 kb, which could be confirmed from Northern hybridization analyses of the isolated rat liver mRNA. One polypeptide of 25 kDa, could be immunoprecipitated with anti-COMT antibody from in vitro translation of rat liver mRNA. Employing the DNA blot analysis only one COMT-encoding gene was found in the rat genome. © 1990.</p>},
  author       = {Salminen, Marjo and Lundström, Kenneth and Tilgmann, Carola and Savolainen, Raija and Kalkkinen, Nisse and Ulmanen, Ismo},
  issn         = {0378-1119},
  keyword      = {genomic library,Methylating enzyme,nucleotide sequencing,recombinant DNA,RNA analysis,λ gt11 expression screening},
  language     = {eng},
  number       = {2},
  pages        = {241--247},
  publisher    = {ARRAY(0xb21da38)},
  series       = {Gene},
  title        = {Molecular cloning and characterization of rat liver catechol-O-methyltransferase},
  url          = {http://dx.doi.org/10.1016/0378-1119(90)90231-F},
  volume       = {93},
  year         = {1990},
}