University of Khartoum

Molecular Characterization of Sudanese Desert Sheep (Ovis aries L, 1758) and Sudanese Nubian Goats (Capra hircus L, 1758) from Gezira and Sinnar States, Sudan

Molecular Characterization of Sudanese Desert Sheep (Ovis aries L, 1758) and Sudanese Nubian Goats (Capra hircus L, 1758) from Gezira and Sinnar States, Sudan

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Title: Molecular Characterization of Sudanese Desert Sheep (Ovis aries L, 1758) and Sudanese Nubian Goats (Capra hircus L, 1758) from Gezira and Sinnar States, Sudan
Author: Mustafa, Saadia Ahmed El Tigani Abd ElGadir
Abstract: This study is designed to investigate the genetic and molecular characterization of the Sudanese Desert sheep (Ovis aries) and the Sudanese Nubian goats (Capra hircus). This research is carried out in two adjacent states, Gezira and Sinnar. Blood samples from sheep (Ovis aries) and goats (Capra hircus) were collected from different sites in Sinnar and Gezira States. A total number of three hundred (300) specimens of sheep (150) and goats (150) from Sinnar State and Gezira State were used for this study. Meat samples were collected from muscle tissue. Milk was extracted for casein and whey protein analysis. Meat and milk proteins were separated using Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS PAGE). Blood was collected from the jugular vein of sheep and goats in 5 ml vacutainer tubes containing ethylene diamine tetera-acetic acid (EDTA). Amplification of the genomic DNA from sheep and goats was performed by PCR using cytochrome b universal primer, for both species. Two specific primers for Leptin (LEP) and Myostatin (MSTN) genes were used.The resulting PCR products were sequenced and the sheep and goat sequences were analyzed using the GenBank nucleotide database. Pairwise distances, phylogenetic trees and protein modeling were done. The translated protein was subjected to Blast P and in silicio tools were used for mutation analysis and prediction of the 3-D structure of wild and mutant type proteins. The results of protein profile banding patterns for milk proteins (casein, whey proteins) showed no variation in milk protein among the members of sheep and goat breeds of the two locations. There was no difference in the meat protein banding pattern as well (P – value ˃ 0, 05). Also, the resulting dendrograms (tree) of the sheep and goat breeds from both Sinnar and Elvi Butana resulted in close relationships of the populations. PCR amplification gave fragments of 357 bp of cytochrome b, 471 bp for leptin and two fragments for myostatin gene with sizes 450 bp and 516 bp. Alignment of cyt b, LEP and MSTN gene sequences with sequences from GenBank database showed high similarity between sheep breeds from Sudan breeds with those from GenBank collection. The same resulted with goat breeds. Nineteen cytochrome b nucleotide sequences of sheep were single nucleotide polymorphisms (SNPs). They were detected in all samples from both locations and were similar to GenBank sheep species. Five cytochrome b SNPs of sheep were novel mutations three of which were represented in all sheep samples while two were unique to two samples from each Sinnar and El Butana. two SNPs were silent or synonymous, the other were transitions and transversions affecting the protein by changing arginine to tryptophan, substitution of glycine with glutamic acid and substitution of leucine with glutamine. However, in goats cytochrome b there were two novel SNPs and synonymous mutations in cyt b of goat (Capra hircus ) in this study , one is unique to Sinnar goat breeds while the other is detected in both Sinnar and El Butana breeds. However, none of the two SNPs is matching GenBank sequences. The phylogenetic tree and analysis of genetic distances of cyt b of Sudanese desert sheep and Sudanese Nubian goats indicated the close relationships between Sinnar and El Butana breeds. Results for leptin gene of sheep from Sinnar revealed two substitutions of arginine to glutamine and similar to Iran goat breeds. MSTN gene of goats resulted in two novel synonymous mutations from Sinnar samples. An interesting result is the presence of five tandem nucleotides (TTTTA) found in all samples of Sinnar goats similar to China breeds. Deletion of thesevii nucleotides was observed in El Butana goats matching reference types in the GenBank. Results of phylogenetic tree and genetic distances of leptin and MSTN genes indicted that all Sudanese sheep and goat breeds are closely related with some similarity to world breeds.
URI: http://khartoumspace.uofk.edu/123456789/27125


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