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Single Nucleotide Polymorphisms And Haplotype Analyses In Tilapia Fish Inferred From MtDNA D-loop And Cyt-b Regions | 97619
ISSN: 2155-952X
Journal of Biotechnology & Biomaterials
Open Access
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Objective: The research was aimed at analysing single nucleotide polymorphisms and haplotypes on D-loop and Cyt-b
regions of the mitochondrial DNA of tilapia fish.
Methods: Fifteen and thirteen tilapia fish were obtained from two populations, South-South (Domita farm)
and South West (Odeda farm). DNA extraction from fish tissue was done using Quick-gDNATM mini prep kit
after which PCR amplification was carried out. Sequencing of the two mtDNA regions were done using forward
primer 5???- GGATTYTAACCCYTRCCCC- 3??? and reverse 3???-AGTAAGTCAGGACCAAGCC-5??? for D-loop and
5???-GGATTTTAACCCTTACCCC-3??? and 3???-AGTAAAGTCAGGACCAAGCC-5??? for Cyt-b region. Statistical analyses
were carried out on the aligned sequenced data using MEGA version 6.06, DnaSP 5.1, Codon code aligner 6.06 as well
as NETWORK 4.6.1.1.
Results: mtDNA polymorphism was highest in the D-loop of South-South (SS) population with 176 polymorphic
sites, while South-West (SW) population had 162 polymorphic sites translating to 176, 162 and 144 SNPs with nonsynonymous
substitutions higher than synonymous substitutions. Haplotype diversities (Hd) were 1.00�0.024 and
1.00�0.030 while nucleotide diversities were 0.168�0.086 and 0.161�0.084 for D-loop of SS and SW populations,
respectively. For Cyt b region, haplotype and nucleotide diversities were 0.91�0.003 and 0.051�0.016. Positive selection
was more on mtDNA D-loop of tilapia sampled from SS than those from the SW as well as Cyt-b region of tilapia fish
from SS. 28 haplotypes were identified among the tilapia from SS and SW with no shared haplotypes while 9 haplotypes
were identified from the Cyt-b region with haplotypes 4, 5, 6 and 7 shared between species. Median joining network
analysis revealed population-based clustering pattern. Demographic expansion was not significant using Tajima???s D and
Fu???s F statistics.
Conclusion: Higher SNPs were revealed in mtDNA D-loop when compared with mtDNA Cyt-b region of tilapia fish.