Evaluation of RAPD-PCR and protein profile analysis to differentiate Vibrio harveyi strains prevalent along the southwest coast of India.
Sixty five isolates of Vibrio harveyi were subjected to random amplified polymorphic DNA (RAPD)-PCR analysis and protein profiling to investigate the genetic variability among V. harveyi prevalent along the coast and also assess the discriminating ability of these two molecular methods. A total of 10 RAPD primers were assayed for their specificity in detecting V. harveyi, of which only two primers: PM3 and CRA25 were highly reproducible and found suitable for use in RAPD-PCR. The genetic diversity among V. harveyi isolates assessed by RAPD-PCR using PM3 primer yielded 35 different RAPD patterns which clustered the isolates into 15 groups at 72% similarity level. Similarly, RAPD-PCR with CRA25 clustered the 38 patterns into 10 groups at 74% similarity. The discriminatory index (D) value calculated for RAPD fingerprints generated with PM3 and CRA25 were 0.90 and 0.85, respectively. On the other hand, molecular typing of V. harveyi using whole cell proteins generated profiles that showed no major difference indicating the technique to be not useful in typing strains of this bacterium. However, a few of the isolates showed the presence of unique band of 28 kDa that needs to be further investigated to understand the role of the protein in disease process if any.
Maiti
Biswajit
B
Department of Fishery Microbiology, College of Fisheries, Karnataka Veterinary Animal and Fisheries Sciences University, Mangalore 575 002, India.
Shekar
Malathi
M
Khushiramani
Rekha
R
Karunasagar
Iddya
I
Karunasagar
Indrani
I
D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods*; D005798:Genes, Bacterial / Q000235:genetics; D006460:Hemolysin Proteins / Q000235:genetics; D007194:India; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*; D012623:Seawater / Q000382:microbiology*; D014733:Vibrio / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D001426:Bacterial Proteins; D006460:Hemolysin Proteins; C073845:hemolysin, Vibrio