Janthinobacterium lividum

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Janthinobacterium lividum is an aerobic, gram-negative, soil-dwelling bacterium . It is a dark purple color and is often found on the skin of amphibians especially frogs (Rebollar).

References:

Rebollar, E. A., Simonetti, S. J., Shoemaker, W. R., & Harris, R. N. (2016, April 4). Direct and Indirect Horizontal Transmission of the Antifungal Probiotic Bacterium Janthinobacterium lividum on Green Frog (Lithobates clamitans) Tadpoles. Retrieved April 5, 2017

Date collected: February 8, 2017

Methods for isolation and identification:

  • Water from the top of Buffalo Creek was collected (Figure 1)
  • The sample was placed onto an augar plate and kept at room temperature
  • Results were taken at 24 hours and 48 hours
  • A purple colony was chosen for 16s RNA sequencing by PCR amplification (Figure 2)
  • The enzyme MSP1 was used to help identify the genus and species.

800px-Map_of_Virginia_highlighting_Prince_Edward_County.svgFigure 1. Collection Site

IMG_1101

Figure 2. Colony collected

 

 

 

 

 

 

 

Results

 

A 1550 bp product was amplified by the PCR (Figure 3).

Slide1

NCBI BLAST analysis revealed 99% match of the 16s rRNA gene of Janthinobacterium lividum (Figure 4).

Janthinobacterium lividum strain DSM 1522 16S ribosomal RNA gene, partial sequence

NR_026365.114691
Alignment statistics for match #1
Score Expect Identities Gaps Strand Frame
1777 bits(962) 0.0() 964/965(99%) 0/965(0%) Plus/Plus
Query  26   TCGAACGGCAGCACGGAGCTTGCTCTGGTGGCGAGTGGCGAACGGGTGAGTAATATATCG  85
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  34   TCGAACGGCAGCACGGAGCTTGCTCTGGTGGCGAGTGGCGAACGGGTGAGTAATATATCG  93

Query  86   GAACGTACCCTGGAGTGGGGGATAACGTAGCGAAAGTTACGCTAATACCGCATACGATCT  145
            ||||||||||| ||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  94   GAACGTACCCTAGAGTGGGGGATAACGTAGCGAAAGTTACGCTAATACCGCATACGATCT  153

Query  146  AAGGATGAAAGTGGGGGATCGCAAGACCTCATGCTCGTGGAGCGGCCGATATCTGATTAG  205
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  154  AAGGATGAAAGTGGGGGATCGCAAGACCTCATGCTCGTGGAGCGGCCGATATCTGATTAG  213

Query  206  CTAGTTGGTAGGGTAAAAGCCTACCAAGGCATCGATCAGTAGCTGGTCTGAGAGGACGAC  265
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  214  CTAGTTGGTAGGGTAAAAGCCTACCAAGGCATCGATCAGTAGCTGGTCTGAGAGGACGAC  273

Query  266  CAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTT  325
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  274  CAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATTT  333

Query  326  TGGACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGAGTGAAGAAGGCCTTCGGGTT  385
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  334  TGGACAATGGGCGAAAGCCTGATCCAGCAATGCCGCGTGAGTGAAGAAGGCCTTCGGGTT  393

Query  386  GTAAAGCTCTTTTGTCAGGGAAGAAACGGTGAGAGCTAATATCTCTTGCTAATGACGGTA  445
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  394  GTAAAGCTCTTTTGTCAGGGAAGAAACGGTGAGAGCTAATATCTCTTGCTAATGACGGTA  453

Query  446  CCTGAAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAA  505
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  454  CCTGAAGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAA  513

Query  506  GCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTTTGTAAGTCTGATGTGA  565
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  514  GCGTTAATCGGAATTACTGGGCGTAAAGCGTGCGCAGGCGGTTTTGTAAGTCTGATGTGA  573

Query  566  AATCCCCGGGCTCAACCTGGGAATTGCATTGGAGACTGCAAGGCTAGAATCTGGCAGAGG  625
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  574  AATCCCCGGGCTCAACCTGGGAATTGCATTGGAGACTGCAAGGCTAGAATCTGGCAGAGG  633

Query  626  GGGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGATATGTGGAGGAACACCGATGGCG  685
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  634  GGGGTAGAATTCCACGTGTAGCAGTGAAATGCGTAGATATGTGGAGGAACACCGATGGCG  693

Query  686  AAGGCAGCCCCCTGGGTCAAGATTGACGCTCATGCACGAAAGCGTGGGGAGCAAACAGGA  745
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  694  AAGGCAGCCCCCTGGGTCAAGATTGACGCTCATGCACGAAAGCGTGGGGAGCAAACAGGA  753

Query  746  TTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCTACTAGTTGTCGGGTCTTAATTGA  805
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  754  TTAGATACCCTGGTAGTCCACGCCCTAAACGATGTCTACTAGTTGTCGGGTCTTAATTGA  813

Query  806  CTTGGTAACGCAGCTAACGCGTGAAGTAGACCGCCTGGGGAGTACGGTCGCAAGATTAAA  865
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  814  CTTGGTAACGCAGCTAACGCGTGAAGTAGACCGCCTGGGGAGTACGGTCGCAAGATTAAA  873

Query  866  ACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCA  925
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  874  ACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGATGATGTGGATTAATTCGATGCA  933

Query  926  ACGCGAAAAACCTTACCTACCCTTGACATGGCTGGAATCCCCGAGAGATTGGGGAGTGCT  985
            ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Sbjct  934  ACGCGAAAAACCTTACCTACCCTTGACATGGCTGGAATCCCCGAGAGATTGGGGAGTGCT  993

Query  986  CGAAA  990
            |||||
Sbjct  994  CGAAA  998
Figure 4. NCBI BLAST

Contributed By:

Taylor Johnson and Sarah Kamen, BIOL 250 Spring 2017