LOCALIZATION OF FLAGELLIN GENES ON THE PHYSICAL MAP OF METHANOCOCCUS-VOLTAE

Methanococcus voltae DNA, digested individually with the restriction e nzymes ApaI, SacII, BamHI, or EagI, was resolved by pulsed-field gel e lectrophoresis reproducing the previously published digestion patterns . Hybridization of a flagellin gene-specific probe to such gels dried down (unblots) resulted in the identification of one band per enzyme h arboring the flagellin genes. These bands all overlapped, revealing th at an approximately 15-kb BamHI/EagI DNA fragment should harbor the fl agellin genes. Double digestion with BamHI and EagI resulted in the re solution of two bands in the 15-kb region of the gel. Separation of th ese two fragments prior to blotting and probing with a flagellar gene- specific probe revealed that one of these fragments possessed the flag ellar sequences. The presence of an EagI restriction site in flaB3 loc alized the flagellin genes precisely at the junction of EagI fragments Ea2 and Ea5 at approximately the 1800-kb position of the physical map .