Work

Molecular cloning and characterization of the gene encoding the DNA methyltransferase, M.CviBIII, fromChlorellavirus NC-1A

Year: 1987

Type: article

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.CviBIII 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.

Source: Nucleic Acids Research

DOI: 10.1093/nar/15.23.9807

Authors: Kenneth E. Narva, Douglas L. Wendell, Merri P. Skrdla, James L. Van Etten

Institution: University of Nebraska–Lincoln

Language: English

Topic: Bacteriophages and microbial interactions

Subfield: Ecology

Field: Environmental Science

Domain: Physical Sciences

Open Access status: bronze

Funders: U.S. Department of Energy, University of California, Davis, National Institute of General Medical Sciences

Awards: G2339259819, G969889393