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Genetic Diversity Analysis Of Sahiwal And Dhanni Cattle Breeds By Cytochrome B Gene And Microsatellite Markers

By: Zahoor Ahmed | Prof.Dr.Masroor Elahi Babar.
Contributor(s): Mr. Tanveer Hussain | Prof.Dr.Muham.
Material type: materialTypeLabelBookPublisher: 2010Subject(s): Institute of Biochemistry & BiotechnologyDDC classification: 1289,T Dissertation note: Pakistan has various dairy breeds of cattle but the genetic data of different cattle breeds including Sahiwal and Dhanni is lacking which need to be established for their genetic identification. Blood samples of unrelated true representative of breeds (Sahiwal and Dhanni) were collected from their respective home tracts and different Government livestock farms. DNA extracted with the standard protocol (Inorganic Method) in Molecular Biology and Genomic Laboratory, Institute of Biochemistry and Biotechnology (IBBT), University of Veterinary and Animal Sciences, Lahore. Nine fluorescent dye labeled microsatellite markers having high polymorphism information content (PIC) values were used and genotyping was done. These results were analyzed statistically by softwares "POPGENE 1.31 and POWER STAT" 2.1. Allele frequency, heterozygosity, homozygosity, polymorphism information content (PIC), power of discrimination, power of exclusion, F-Statistics and Gene Flow for all Loci, population's dendogram, Nei's genetic identity and genetic distance/ diversity were calculated. The average observed heterozygosity was 0.5845 and 0.5911 in Dhanni and Sahiwal respectively. The mean observed homozygosity was 0.4155 and 0.4089 in Dhanni and Sahiwal respectively. The average PIC (Polymorphic Information Content) values of nine loci showed by Dhanni and Sahiwal cattle are 0.61 and 0.77 respectively. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between Dhanni and Sahiwal cattle breeds. For further confirmation of the breeds amplification of the mitochondrial Cyt b gene was done with especially designed primers which were designed by using Primer3 software. Sequencing of PCR fragments was done. Analysis of the sequences was performed by multiple sequence alignment with the help of Blast 2sequence and BioEdit soft wares. Identified SNPs were analyzed and haplotypes were formed. Phylogenetic tree was constructed by MEGA 4.1. The use of genetic markers provided the information on population genetic structures of the indigenous cattle breeds even if they lack detailed pedigree recording data. The study on the genetic diversity showed the differentiation of breeds and individual breeds have unique combinations of genes as a result of phylogenetic tree. This work will provide the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in Pakistan in future according to FAO global Farm Animal Genetic resource data.
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Pakistan has various dairy breeds of cattle but the genetic data of different cattle breeds including Sahiwal and Dhanni is lacking which need to be established for their genetic identification. Blood samples of unrelated true representative of breeds (Sahiwal and Dhanni) were collected from their respective home tracts and different Government livestock farms. DNA extracted with the standard protocol (Inorganic Method) in Molecular Biology and Genomic Laboratory, Institute of Biochemistry and Biotechnology (IBBT), University of Veterinary and Animal Sciences, Lahore. Nine fluorescent dye labeled microsatellite markers having high polymorphism information content (PIC) values were used and genotyping was done. These results were analyzed statistically by softwares "POPGENE 1.31 and POWER STAT" 2.1. Allele frequency, heterozygosity, homozygosity, polymorphism information content (PIC), power of discrimination, power of exclusion, F-Statistics and Gene Flow for all Loci, population's dendogram, Nei's genetic identity and genetic distance/ diversity were calculated. The average observed heterozygosity was 0.5845 and 0.5911 in Dhanni and Sahiwal respectively. The mean observed homozygosity was 0.4155 and 0.4089 in Dhanni and Sahiwal respectively. The average PIC (Polymorphic Information Content) values of nine loci showed by Dhanni and Sahiwal cattle are 0.61 and 0.77 respectively. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between Dhanni and Sahiwal cattle breeds. For further confirmation of the breeds amplification


of the mitochondrial Cyt b gene was done with especially designed primers which were designed by using Primer3 software. Sequencing of PCR fragments was done. Analysis of the sequences was performed by multiple sequence alignment with the help of Blast 2sequence and BioEdit soft wares. Identified SNPs were analyzed and haplotypes were formed. Phylogenetic tree was constructed by MEGA 4.1.
The use of genetic markers provided the information on population genetic structures of the indigenous cattle breeds even if they lack detailed pedigree recording data. The study on the genetic diversity showed the differentiation of breeds and individual breeds have unique combinations of genes as a result of phylogenetic tree. This work will provide the genetic data which will be helpful in breed identification and making effective breeding policies and conservational activities in Pakistan in future according to FAO global Farm Animal Genetic resource data.

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