Pseudomonas koreensis are coccus bacteria with Gram-negative cells that were isolated from Korean agricultural soils . These bacteria cells are circular with whitish-yellowish coloring on Luria-Bertani (LB) agar.
- Kwon S, Kim J, Park I, Yoon S, Park D, Lim C, Go S. 2003.Pseudomonas koreensis sp. nov., Pseudomonas umsongensis sp. nov. and Pseudomonas jinjuensis sp. nov., novel species from farm soils in Korea. International Journal of Systematic and Evolutionary Microbiology.53.21-27
Data Collected: February 2,2017
Methods for isolation and identification:
- Five grams of wet and dry soil were collected from Buffalo Creek, located in Lancer Park (Figure 1). Each sample was placed into a sterile tube and 25mL of water was added. Ten milliliters was pipetted from each tube into a second sterile tube, and ten milliliters was pipetted out again and placed into a third sterile tube. The samples were spread across a sterile agar plate, and incubated at 37 degrees Celsius for 16 hours.
- A mucus yellow colored colony (Figure 2) was singled out for the 16S rRNA gene sequencing by PCR amplification.
- The PCR product was digested and sequenced in order to classify the species of the bacteria.
Figure 1: Site of Collection
Figure 2. Colony selected for identification
Results of PCR product and MSP1 digestion
MSP1 Digestion (Figure 3): The amplification of our PCR Product was about 950 bp. When MSPI digest was added, a band at 500 bp appeared.
NCBI BLAST analysis of colony 7-W
Sequence Analysis (Figure 4): The PCR products produced 908 “good reads”. The chromatogram sequence is available for comparison as a pdf (7W_PREMIX_KG2505_5 (1)). From the product, a NCBI BLAST analysis was conducted and generated a sequence with 908 identical base pairs out of 910, with a match of 99%. These reads helped find what bacteria it was by identifying the genus and species.
Contributed by: Megan A. Curry, Debbie N. Pitt, & Brittany N. Barton, BIOL 250 Spring 2017, Group 7