S-STEM Scholar Jacye Thompson's Blog

During this week Deinococcus Caeni’s susceptibility to tetracycline was still being tested. It has been confirmed that this bacteria strain does have an efflux pump just as Deinococcus Aquaticus did. However, the concentrations used last week were 0 ug/ml, 10 ug/ml, 50 ug/ml, and 100 ug/ml. We expected growth on at least the 10 ug/ml plate, but that concentration was bactericidal. The concentrations were reduced to 0 ug/ml, 1 ug/ml, 2 ug/ml, and 6 ug/ml.  Materials: Liquid culture of D. Caeni Four flasks of 25 ml TGY with Agar Tetracycline with stock concentrations of 2.5 mg/ml Hot plate Vortex Micropipette and corresponding tips Procedures: Inoculate flask of 10 ml TGY with colony from D. Canei plate Place in shaking incubator for 72 hours Autoclave four flask of 25 ml TGY with 1.5% agar to pour plates Reheat the sterilized agar on a hot plate Let cool to room temperature before adding antibiotic  Remove TGY, add tetracycline, add 250 ul of bacteria and vortex at level six fo...

Currently in the lab Deinococcus Canei is being sequenced. Though the genome has not yet been fully sequenced, it can be said that D. Canei is closely related to Deinococcus Aquaticus. With that, D. Canei susceptibility is tested this week to determine if it has an efflux pump, and if so, identify how useful it is against the antibiotic tetracycline. In the last test with D. Aquaticus the concentrations 0 ug/ml, 10 ug/ml, 100 ug/ml, and 1000 ug/ml. The only plate that had growth was the 0 ug/ml, the only plate that did not contain tetracycline. Since, these concentrations were bactericidal the concentrations used for this experiment were lowered to 0 ug/ml, 10 ug/ml, 50 ug/ml, and 100 ug/ml. Materials:  Liquid culture of D. Canei  Four flasks of 25 ml TGY with agar Tetracycline with stock concentrations of 2.5 mg/ml Hot plate Vortex Micropipette and corresponding tips Procedures: Inoculate a flask of 10 ml TGY with a colony from D. Canei plate Place in shaking incubator for 72...

Deinococcus Aquaticus suseptability to tetracycline was tested by streaking agar plates coated with a soft agar containing increasing levels of tetracycline; 0 ug/ml, 10 ug/ml, 100 ug/ml, and 1000 ug/ml. As states previously, Deinococcus Aquaticus has an efflux pump embedded in its cell membrane. This pump works with the two genes tetA and tetR. The first gene tetA codes from the pump itself, while the second tetR regulates the function of the pump. With this structure in D. Aquaticus’s membrane suggests that it may have a resistance to the antibiotic tetracycline. Tetracycline is a protein that synthesis inhibitor, preventing the aminoacyl-tRNA from bringing to the active site of mRNA. This stops the bacteria from growing and replicating.   Materials:  ·        Solid Agar Plates ·        Soft agar ·        Liquid Culture of D. Aquaticus ·        Hot pla...