Pseudomonas brenneri strain CFML97-391 16s ribosomal. Pseudomonas brenneri strain is a gram-negative bacterium designated strain H3-R18T that was isolated from seashore sand collected from Homi Cape, Pohang City, Korea. Cells were Gram-negative, aerobic, non-motile, cream-coloured, mesophilic and slightly halotolerant (Weon, 2006).
Baida N, Yazourh A, Singer E, Izard D. Pseudomonas brenneri sp. nov., a new species isolated from natural mineral waters. Research in microbiology. [Internet]. 2001; [cited 2017 April 2]; 5: 493-502: Available from http://www.sciencedirect.com/science/article/pii/S0923250801012232?via%3Dihub
Weon H-Y. Pseudomonas pohangensis sp. nov., isolated from seashore sand in Korea. International Journal Of Systematic And Evolutionary Microbiology. 2006 [cited 2017 Apr];56(9): 2153–2156: Available from https://www.ncbi.nlm.nih.gov/pubmed/16957113
Date Collected: February 8, 2017
Methods for isolation and identification:
- Fifty milliliters of surface water was collected using conical tube from the surface of a certain location on Buffalo Creek (Figure 1). A sterile swab was mixed in the sample and spread across a sterile nutrient agar plate. The plate was incubated at 23 degrees Celsius for 48-hours and monitored for change.
- A white colony from each sample (Figure 2) was selected for 16S rRNA gene sequencing by PCR amplification.
- The PCR product was digested with MspI and sequenced to identify the genus and species of the bacteria.
Figure 1. Site of collection (on Buffalo Creek)
Figure 2. Colony selected for identification
- MspI digestion (Figure 3): A 1500 bp product was amplified by PCR. Upon digestion with MspI, two bands were observed at approximately 500 bp and 100bp.
Figure 3. Results of PCR amplification and MspI digestion
- Sequence analysis (Figure 4): The sequenced PCR product generated 1000 bases of high-quality reads that were used to identify the genus and species of the colony. The chromatogram of the sequence is available as a pdf (PDF of Water Sample). NCBI BLAST analysis revealed 99% identity with bases 21-1019 of the 16s rRNA gene of Pseudomonas brenneri.
Figure 4. NCBI BLAST analysis of the water bacterial colony
Contributed by: Christina Gonzalez and Meagan St. John, BIOL 250 Spring 2017