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Parentage Analysis And Breed Characterization Of Dogs By Microsatellite Markers

by Muhammad Sajid Tahir | 5/31/2010 | Mr.Tanveer Hussain | Mrs.Saeeda.

Material type: book Book; Format: print Publisher: 1990Dissertation note: Pakistan has vast population of dogs belonging to different breeds. Most of the dogs have no pedigree record which is a great threat to conservation of different breeds. No study on DNA fingerprinting of dogs has been conducted in Pakistan. DNA fingerprinting of dogs is necessary to overcome the problems like forensic cases, sale & purchase, individual identity in case of fertilization by more than one male and ownership disputes. Microsatellite markers have been proved as an efficient and powerful tool for parentage testing and breed characterization of dogs. In this study, a panel of microsatellite markers, having high polymorphism information content (PlC) values, was developed. Blood samples were taken from cephalic vein of two breeds of dogs (German shepherd and Labrador retriever). DNA was extracted by Inorganic method. Primers of microsatellite markers were optimized for successful amplification conditions in the Bio-Rad thermocycler. Multiplex PCR was performed, for amplification of these microsatellite markers on 46 samples belonging to 20 families. Genotyping analysis was performed for the PCR products of microsatellite markers on non denaturing polyacrylamide gel. These results were analyzed statistically software "POPGENE 3.3 and POWER STAT". Allele frequency, heterozygosity, homozygosity, polymorphism information content (PlC), power of discrimination and power of exclusion of all microsatellite markers were calculated. Average power of discrimination among non parents, average hetrozygosity, average observed homozygosity and average polymorphism information content (PlC) value for all alleles was 0.809, 0.6345, 0.2913 and 0.724 respectively. Moreover combined power of exclusion reached a significant value of 0.9998. Almost all of the microsatellite markers showed significant variations in both German shepherd and Labrador retriever breeds. Microsatellite "REN41D2Ob" showed maximum variation i.e. 17 alleles and microsatellite"REN49F22b" showed the least variation among all microsatellite markers i.e. 4 alleles. Genotyping results of microsatellite markers were clearly different for two different breeds showing a distinct genetic distance between German shepherd and Labrador retriever breeds. Results of this study lead to development of a panel of microsatellite markers which can be used for parentage analysis and breed characterization of dogs. This was a preliminary study on dogs in Pakistan. This facility can be provided on commercial basis to pet owners and kennel clubs. Moreover this study can become the basis for further research investigations in canines in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1090,T] (1).

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Horses Parentage Analysis And Breed Characterization By Microsafellipe Markers

by Javed Iqbal | Prof.Dr.Masroor Elahi Babar | Dr.Ahmad Ali | Mrs.Saeeda Kalsoom.

Material type: book Book; Format: print Publisher: 2002Dissertation note: Horses (Equus caballus) have been considered one of the most significant domesticated animals in human use, including sports, urban and rural transportation and military logistics. Paternity confirmation and breed identification is the basic pre-requisite for rearing specific breeds and maintaining the pedigree records of horses. DNA finger printing has been successfully used for paternity and breed confirmation. Mainly DNA finger printing is done through microsettalite markers for paternity analysis and breed characterization. The aim of this study was to develop and apply a panel of microsettalite markers for paternity analysis and selected horse breeds characterization. Three horse breeds Percheron, Pak Arab and Thoroughbred were selected for this purpose. Sampling of 20 families (Foals, Mare and Stallion) of three selected horse breeds was conducted from Remount Depot Mona. The blood samples was collected in sterilized Falcon tubes each containing lOOjiL EDTA (0.2 mM). DNA was extracted from all blood samples by using inorganic method. The microsatellite markers were selected from ISAG (International Society of Animal Genetics), Indian and TKY recommended panels. The primers were designed for these microsatellite markers using primer3 free ware. All Microsatellite markers used were direpeats. Conditions for successful amplification by PCR (Polymerase Chain Reaction) were optimized and microsatellite markers were grouped into 6 multiplexes (tn and diplex). PCR products of optimized multiplexes were visualized on U V illuminator after gel electrophoresis. DNA amplification was done through PCR by using optimized multiplexes of microsatellite markers for all the samples. Polyacrylarnide Gel Electrophoresis (PAGE) was used for genotyping of amplified DNA samples. Allele sizes of all amplicons were calculated by relative flow method. Alleles for all microsatcilite markers were analyzed statistically by "POPGEN 32 and POWER STAT" software. The investigation revealed average PlC value 0.97, average observed heterozygosity 0.923 1, average observed homozygosity 0.0769, combined power of exclusion (P.E) 0.9999 and combined polymorphic loci percentage for 13 microsettalite markers was 100%. Perchron breed showed genetic identity with Pak Arab and Thoroughbred up to 0.3470 and 0.6157 respectively. Pak Arab exhibited genetic identity with Thoroughbred horse up to 0.6616 where as Perchron breed showed genetic distance with Pak Arab and Thoroughbred up to 1.0585 and 0.4850 respectively. Pak Arab exhibited genetic distances with Thoroughbred up to 0.413 1. Results of analysis were used to describe the new microsettalite markers assay for parentage confirmation and breed characterization of selected breeds (Perchron, Pak Arab and Thoroughbred) horses. 'Ihis panel of microsetallite marker was useful and reliable tool for individual identification and parentage analysis in horses. This microsettalite marker panel can be used on commercial basis in Pakistan. Availability: Items available for loan: UVAS Library [Call number: 1092,T] (1).

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Characterization Of Dgati Gene For K232A Polymorphism In Pakistani Cattle Breeds

by Rashid Hussain | Prof.Dr.Masroor Elahi Babar | Dr.Aftab Anjum | Mrs.Saeeda Kalsoom.

Material type: book Book; Format: print Publisher: 2011Dissertation note: Milk is a balanced diet because it contains all the essential nutrients like carbohydrates, fats, proteins, vitamins, minerals and enzymes required for health. In these milk nutrients fat is second most important component of milk. Primary component of milk fat is triglycerides (triacylglycerols or TAG), a typical storage form of lipids. DGAT1 gene has important role in milk fat percentage. This gene is located on the centromeric end of the bovine chromosome 14. It was recently studied that nonconservative dinucleotide (AA?GC) substitution in exon 8 of DGAT1 gene, change lysine to alanine at position 232 (K232A mutation) of the encoding protein. Animals that have K (Lysine) at position 232 in amino acid sequence of DGAT 1 will have high fat percentage and low milk yield as compared to animals that have A (Alanine) at this position. The objectives of this study were to identify K232A (Lysine232?Alanine) polymorphism in DGAT1 gene in Pakistani cattle breeds. To find relationship between milk production and K232A polymorphism. Blood samples were collected from different Govt. livestock farms/experimental stations. DNA was extracted by organic method. Specific primers were used for the amplification of exon8 of DGAT1 gene. After DNA amplification by Polymerase Chain Reaction, restriction digestion was done by using CfrI enzyme. Fourty animals belonging to Sahiwal and Dhanni breeds were genotyped for K232A polymorphism by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) technique. Sequencing was carried out to confirm the results of restriction digestion. From the data analyzed it was observed that all the animals of Sahiwal and Dhanni breeds had K allele and no A allele was identified. It has been shown that a missense mutation (Lys232 ? Ala) in the bovine DGAT1 gene in Pakistan cattle breeds is not common. Availability: Items available for loan: UVAS Library [Call number: 1283,T] (1).

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