Strains and Genes that Facilitate Genetic Manipulation of Hyperthermophiles

Innovative Solutions to Global Needs TECHNOLOGY CASE 1576 USPTO Links Introduction It would be difficult to overestimate the contribution of genetic manipulation to the study of any biological system, and it is an essential tool for the metabolic engineering of biosynthetic and substrate utilization pathways. This is particularly true for the archaea since, in spite of their environmental and industrial importance, coupled with their unique molecular features, much remains to be learned about their biology. The marine hyperthermophilic anaerobe Pyrococcus furiosus is of special interest not only for its ability to grow optimally at 100° C and the implications of this trait for its biology but also for industrial applications of its enzymes, as well as its capacity to produce hydrogen efficiently in a highly efficient manner. The development of genetic systems in the archaea, in general, presents many unique challenges given the extreme growth requirements of many of these organisms. To date, genetic systems of various levels of sophistication have been developed for representatives of all major groups of archaea, including halophiles, methanogens, thermoacidophiles, and hyperthermophiles. One of the most significant barriers to genetic manipulation of archaea, in general, and hyperthermophiles, in particular, is the lack of selectable markers. Antibiotic selection strategies used in mesophilic bacteria are typically ineffective because the molecular machineries of archaea are not affected by the antibiotic.