Determination of genome size and a preliminary physical map of an extreme alkaliphile, Micrococcus sp. Y-1, by pulsed-field gel electrophoresis.
Large restriction fragments of genomic DNA from Micrococcus sp. Y-1 were separated by pulsed-field gel electrophoresis (PFGE). Since Micrococcus sp. Y-1 has a G+C content of approximately 70%, restriction fragments were obtained by digesting chromosomal DNA with endonucleases which recognize A+T-rich sequences. Five enzymes, SspI, SpeI, XbaI, HpaI and EcoRI, were used for generation of distinctly separated fragments in the size range 100-500 kb. No site for DraI was detected. In contrast, sites for 8-base-recognizing enzymes, but not for NotI and SfiI, were frequent. The genome size of Micrococcus sp. Y-1 was determined from restriction fragments separated by PFGE, and was estimated to be approximately 4061 kb. Partial digestion experiments revealed the order of the six SspI fragments on the chromosome.
Park
J H
JH
Genome Program, Genetic Engineering Research Institute, KIST, Taejon, Korea.
Song
J C
JC
Kim
M H
MH
Lee
D S
DS
Kim
C H
CH
D000468:Alkalies / Q000378:metabolism; D001483:Base Sequence; D002874:Chromosome Mapping; D004269:DNA, Bacterial / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D016680:Genome, Bacterial; D008837:Micrococcus / Q000235:genetics* / Q000378:metabolism; D015183:Restriction Mapping
D000468:Alkalies; D004269:DNA, Bacterial