S-STEM Scholar Jacye Thompson's Blog

There are over 100 species in the Deinococcus genus that have been identified . Interest in this genus has grown as they are polyextremophiles , including the phenotype of radio resistance, and have potential bioremediation genes. These genes are capable of the degradation of complex waste mixtures, org anic solvents, heavy metals and radioisotopes. For example, the Deinococcus radiodurans is the most characterized and has attracted interest for degrading radioisotope. Deinococcous geothermalis is another species that has been under the scope for removal of radioactive waste in highe r temperatures. However, there is a limited amount of research that has been conducted on other species . As of now only a hand full of species have been successfully transformed, a sta ndard laboratory technique used to integrate exogenous DNA .   Deinococcus radio durans was the first to be transformed . However, the transformation protocol requires a microgram quantity of the speci...

  This week concludes the Deincoccus Caeni’s susceptibility to the tetracycline test. The concentrations used during the last experiment were 0 ug/ml, 1 ug/ml, 2 ug/ml, and 6 ug/ml. These concentrations were still too high to see any growth on the plates with antibiotics so the concentrations used were 0 ug/ml, 0.25 ug/ml, 0.5 ug/ml, and 0.75 ug/ml.  Materials: Liquid culture of D. Caeni Four flasks of 25 ml TGY with Agar Tetracycline with a stock concentration of 2.5 mg/ml Hot plate Vortex Micropipette and corresponding tips Procedures: Inoculate a flask of 20 ml TGY with a colony from the D. Canei plate Place in a shaking incubator for 72 hours Autoclave four flaks of 25 ml TGY with agar to pour plates Reheat the sterilized agar on a hot plate Let cool to room temperature before adding the antibiotic  Remove TGY, add tetracycline, add bacteria then vortex for six seconds on level six, pour in labeled petri plates 0 ug/ml: remove 0 ul TGY, add 250 ul of bacteria 0.25 ug/...

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...

There are a vast number of bacteria species in the environment, the species in focus is Deinococcous.     The interest in Dienococcous has come about because of its desirable characteristics. The Dienococcous species are the most radiation-resistant micro-organisms that have been discovered and have genes that encode cell cleaning systems that eliminate organic and inorganic cell toxins (Gerber, 2015). This demonstrates it capacity to persist under multiple stressors and the efficiency to repair DNA damage. There is still much to be learned about the species of Deinococcous, thus this project will be testing Deinococcous Aquaticus’s susceptibility to the antibiotic tetracycline.   Deinococcous Aquaticus is a gram-negative bacterium, so it appears pink under a microscope after a gram stain (BacDive, 2023). Gram-negative describes the structure of the cell wall. It appears pink because the peptidoglycan cannot retain the crystal violet, the first dye added, because it is un...