Widger, William R.Housammy, Zina2018-02-272018-02-272017-10-12http://hdl.handle.net/10657/2488Dormancy is a mechanism by which bacteria are viable but do not grow, called a Viable But Not Culturable (VBNC) state. Our lab has identified 18 distinct proteins that are up-regulated during dormancy; in order to further study these, we want to generate knock-out mutations in Micrococcus luteus and reintroduce mutated genes into the Micrococcus luteus genome. To this end, my project is to use the pigment operon (genes: crtE, B, I, E2, Yg and X) and use the first gene (crtE) as a neutral site to insert mutated gene sequences for analysis. I will describe here the amplification and cloning methods that I am utilizing to use the crtE gene as a neutral site for the insertion of mutated DNA into the Micrococcus luteus genome. For example, we want to insert green fluorescent protein into the neutral site to use as a biomarker for dormancy and resuscitation. This provides a rapid method for measuring the degree of resuscitation based off the return of the green fluorescence.en-USThe author of this work is the copyright owner. UH Libraries and the Texas Digital Library have their permission to store and provide access to this work. Further transmission, reproduction, or presentation of this work is prohibited except with permission of the author(s).Poster