Open AccessA Nhe I macrorestriction map of the Neisseria meningitidis B1940 genome
Author(s) -
Bautsch Wilfried
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06029.x
Subject(s) - restriction enzyme , genome , biology , genetics , pulsed field gel electrophoresis , neisseria meningitidis , restriction map , genomic dna , neisseria , genome size , dna , microbiology and biotechnology , gene , plasmid , bacteria , genotype
A macrorestriction map of the Neisseria meningitidis strain B1940 genome was constructed by two‐dimensional pulsed‐field gel electrophoresis (2D‐PFGE) techniques. Digestion of the genomic DNA with the restriction endonuclease NHe I revealed 15 fragments between 10 kb and 450 kb. The sum of the fragments and resolution of the linearized chromosome yielded a total genome size of about 2.3 Mbp. By overlapping methylation with the Alu I‐methylase six Nhe I recognition sites could be blocked. Fragments were ordered by partial/complete 2D‐PFGE of genomic DNA with and without prior Alu I methylation, respectively. All nine Alu I‐methylase/ Nhe I and 14 Nhe I restriction sites could be mapped on a single circular chromosome. This map will serve as a useful tool for further genetic analysis of meningococci and exemplifies the power of non‐radioactive 2D‐PFGE techniques to construct large physical genome maps with a single restriction enzyme.