Bacillus marisflavi colony 16-1

Bacillus marisflavi is a gram-positive motile bacteria that was previously isolated by Yoon and colleagues from the sea water of a tidal flat of the Yellow Sea in Korea.

References:

  1. Yoon J, Kim I, Kang K, Oh T, Park Y. Int J Syst Evol Microbiol 53(5):1297-1303 doi:10.1099/ijs.0.02365-0

Date collected: December 12, 2016

Methods for isolation and identification:

  • Five milliliters of surface water was collected from the surface of a retention pond in Lancer Park (Figure 1). A sterile swab was mixed in the water and spread across a sterile LB agar plate. The plate was incubated at 37 degrees Celsius for 16 hours.
  • A light yellow colony (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.

 Screen Shot 2017-03-20 at 12.44.54 PMFigure 1. Site of collection

Colony 1 B marisflavi

Figure 2. Colony selected for identification

 

Results:

  • MspI digestion (Figure 3): A 1,500 bp product was amplified by PCR. Upon digestion with MspI, four bands at approximately 650 bp, 400 bp, 225 bp, and 175 bp were identified.

Bacillus marisflavi colony 16-1

Figure 3. Results of PCR amplification and MspI digestion

  • Sequence analysis (Figure 4): The sequenced PCR product generated 857 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 (rDNA_seq_1_PREMIX_KG2496_1). NCBI BLAST analysis revealed 99% identity with bases 35-891 of the 16s rRNA gene of Bacillus marisflavii (Figure 4). Screen Shot 2017-02-15 at 10.41.35 AMFigure 4. NCBI BLAST analysis of colony 16-1

 

 

Contributed by: Erin K. Shanle, BIOL 250 Spring 2017, Group 16