Biochemistry and genetics of autotrophy in Methanococcus. Progress report

Technical Report · Tue Sep 01 04:00:00 EDT 1998

DOI:https://doi.org/10.2172/663534· OSTI ID:663534

Whitman, W B

In the last two years of this research, the most exciting results have come from the work on the genetics of methanococci. First, the author demonstrated that the cryptic plasmid from Methanococcus maripaludis C5, pURB500, could be transformed into Methanococcus maripaludis JJ. Strain JJ is the type strain of M. maripaludis and has only about 65% DNA:DNA hybridization to strain C5. Because of the low relatedness of these strains, it was not obvious that pURB500 could be transferred between them. This goal was achieved by first transforming strain C5 with a series of suicide plasmids containing the pac cassette, which possessed the selectable puromycin resistance marker, and different cloned fragments of pURB500. From the puromycin-resistant transformants, a plasmid was isolated that transformed strain JJ. However, when this plasmid was electroporated into E. coli, only rearrangement products were obtained that contained small portions of the original pURB500. These plasmids no longer transformed Methanococcus. While these experiments did not yield a shuttle vector, they demonstrated that pURB500 could replicate in strain JJ.

• Research Organization: Univ. of Georgia, Athens, GA (United States)
• Sponsoring Organization: USDOE Office of Energy Research, Washington, DC (United States)
• DOE Contract Number: FG05-94ER20158
• OSTI ID: 663534
• Report Number(s): DOE/ER/20158--T1; ON: DE98007374
• Country of Publication: United States
• Language: English

Related Subjects

55 BIOLOGY AND MEDICINE
BASIC STUDIES
BIOCHEMISTRY
DNA HYBRIDIZATION
DNA REPLICATION
GENETICS
METHANOGENIC BACTERIA
PLASMIDS
PROGRESS REPORT