Starzyk, Marvin J.
M.S. (Master of Science)
Department of Biological Sciences
Thermus aquaticus; Plasmids--Genetics; Bacterial genetics; Thermophilic bacteria
Plasmid DNA from Thermus acmaticus YT-1 was obtained through the use of a modified alkaline lysis procedure followed by separation on, and extraction from, low melting point agarose. The 8.7 kb agarose gel plasmid band previously designated pTAl was subjected to restriction enzyme digestion. Analysis of the resulting restriction fragments revealed this plasmid band actually represents two comigrating plasmids, each with a distinct restriction map. This brings the total number of plasmids reported in this species to five. These plasmids have been redesignated pTAl (8.7 kb), pTA2 (8.7 kb), pTA3 (13.7 kb), pTA4 (15.8 kb), and pTA5 (17.5 kb). Restriction Endonuclease Pst I fragments of pTAl and pTA2 were ligated to the Escherichia coli vector pUC18. Transformation of E. coli DH5a and subsequent selection produced 23 recombinant colonies. Twenty of these strains were screened with respect to plasmid content. One of the recombinant strains which harbored a plasmid containing a 7.3 kb fragment of pTA2 was tested for production of thermostable proteins. No thermostable protein was detected by the methods used to identify such a product.
Cooper, Andrew J., "Physical analysis and partial cloning of two co-migrating plasmids from Thermus aquaticus YT-1" (1993). Graduate Research Theses & Dissertations. 4384.
vi, 91 pages
Northern Illinois University
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