Investigation of Pressure Regulation in an Archaebacterial Enzyme
Final rept. 1 Feb 91-31 Jan 94,
PITTSBURGH UNIV PA
Pagination or Media Count:
Initial studies focused on the purification of the hydrogenase from Methanococcus jannaschii. The hydrogenase was characterized, N-terminal sequenced, and its amino acid content was compared to other hydrogenases. Researchers then constructed a library of genomic DNA from Methanococcus jannaschii. The genomic library, with over 1 million independent representatives, was cloned into bacteriophage lambda. The DNA from the organism has been found to be methylated, preventing classic enzyme treatments for gene library synthesis. A gene library has been synthesized by mechanical shearing, and the fragment size is approximately 8-15 kb. Another driving force for the production of the library was the independent discovery of a remarkably thermostable protease which retains activity at 135 deg C. The protease was partially purified. The partially purified sample was used for the studies to determine which class of protease it may be. Lambda, Protease archaebacterial, Enzyme, Hydrogenase, Bacteriophage, DNA.
- Genetic Engineering and Molecular Biology