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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2018: 95.39
For more details click here

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Original Research Articles

PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2017.6(1): 569-578
DOI: http://dx.doi.org/10.20546/ijcmas.2017.601.070


Restriction Sites Annotation on Klebsiella 16s Ribosomal RNA using Selected Sticky End Cutting Endonucleases
Alade Tolulupe* and Arikekpar Ibemologi
Department of Medical Laboratory Science, Faculty of Basic Medical Sciences,College of Health Sciences, Niger Delta University, Wilberforce Island,Amassoma, P.M.B. 071, Bayelsa State, Nigeria 
*Corresponding author
Abstract:

The 16S rRNA gene is composed of highly conserved, specie-specific sequences between different species of bacteria  and the application of restriction endonucleases on the amplified 16S rRNA gene is a novel diagnostic tool  in molecular characterization of bacterial isolates. In a study aimed at characterizing bacteria using some selected endonucleases, a total of 31 isolates collected from the stock culture in the Department of Medical Laboratory Science were re-identified using standard microbiological procedures as Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli and Alcaligenes faecalis. The isolates were subjected to antibiotic susceptibility testing to ascertain their susceptibility pattern using the agar disc diffusion method; followed by DNA extraction, purification, amplification, gel electrophoresis and sequencing.  The Sequences were edited using trace edit, Blasted and Restriction site annotation was carried out using Geneious. Antibiotic susceptibility testing results showed that K. pneumoniae is susceptible to Streptomycin, Ciprofloxacin, Gentamicin, Nalidixic Acid, Pefloxacin, Septrin, Ofloxacin and resistant to Augmentin, Ampicillin, Cephalosporin. Restriction site annotation of the sequences revealed that Xma1 cuts Klebsiella pneumoniae and Pseudomonas spp. at position 499 while EcoR1cuts both Klebsiella pneumoniae and Bacillus spp. at position 561; showing that both Xma1 and EcoR1 cannot be used as restriction markers to differentiate between Klebsiella pneumoniae and Pseudomonas spp. and Bacillus spp. respectively.


Keywords: Klebsiella 16srRNA, restriction site annotation, Sticky ends cutting endonucleases.
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How to cite this article:

Alade Tolulupe and Arikekpar Ibemologi. 2017. Restriction Sites Annotation on Klebsiella 16s Ribosomal Rna using Selected Sticky End Cutting Endonucleases.Int.J.Curr.Microbiol.App.Sci. 6(1): 569-578. doi: http://dx.doi.org/10.20546/ijcmas.2017.601.070