Molecular cloning and characterization of the gene encoding the DNA methyltransferase, M.CviBiii, from Chlorella virus NC-1A

Kenneth E. Narva, Douglas L. Wendell, Merri P. Skrdla, James L. van Etten

Research output: Contribution to journalArticle

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Abstract

The gene encoding the DNA methyltransferase, M.CviBIII, from Chlorella virus NC-1A was cloned and expressed in E. coli plasmid pUC8.Plasmid (pNC-1A.14.8) encoded M.CviBIII methylates adenine in TCGA sequences both in vivo in E.coli and in vitro.Transposon Tn5 mutagenesis localized the M.CviBIII functional domain to a 1.5 kbp region of pNC-1A.14.8 and also indicated that a virus promoter directs transcription of the gene in E. coli. The 2.1 kbp insert containing the M.C.ViBIII gene was sequenced and a single open reading frame of 1131 bp was identified within the domain determined by Tn5 mutagenesis. When the M.CviBIII gene was fused in-frame with the 19 amino-terminal codons of lacZ a 45 kD polypeptide was identified in maxicells as predicted by the DNA sequence. The M.CviBIII gene was not essential for virus replication since a virus M.CviBIII deletion mutant also replicated in Chlorella.

Original languageEnglish (US)
Pages (from-to)9807-9823
Number of pages17
JournalNucleic acids research
Volume15
Issue number23
DOIs
StatePublished - Dec 10 1987

Fingerprint

Chlorella
Gene encoding
Cloning
Methyltransferases
Molecular Cloning
Viruses
Virus
Encoding
DNA
Genes
Escherichia coli
Gene
Mutagenesis
Escherichia Coli
Plasmids
DNA sequences
Polypeptides
Adenine
Transcription
Virus Replication

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Molecular cloning and characterization of the gene encoding the DNA methyltransferase, M.CviBiii, from Chlorella virus NC-1A. / Narva, Kenneth E.; Wendell, Douglas L.; Skrdla, Merri P.; van Etten, James L.

In: Nucleic acids research, Vol. 15, No. 23, 10.12.1987, p. 9807-9823.

Research output: Contribution to journalArticle

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