Abstract:
Several breeds of buffalo and cattle in Pakistan show mark differences in the milk yield.
These animals are in a range of high milk producer to low. A proportion of these
differences are explained by the animal genetic make up. In the earlier studies, a number
of DNA polymorphisms have been reported in candidate genes. Among these few of
candidate genes studied in the present study, are bovine growth hormone (bGH), kappa-
casein (k-CN) and beta-lactoglobulin (B-LG) genes using PCR-RFLP methodology. Two
genetic variants (A and B) and three genotypes AA, AB and BB have been identified.
The genotyping of bovine growth hormone (bGH), kappa casein (-CN) and beta-
lactoglobulin (β-LG) alleles is of practical importance since bGH L, -CN A & B and β-
LG B alleles were found to be correlated with commercially valuable parameters.
Genotype information on 366 animals from four breeds including one buffalo and three
cattle breeds of Pakistan regarding polymorphisms located in the candidate genes was
compared to literature reports. The results from frequency analysis revealed minor
deviations from previous reports. Allele frequencies were calculated by gene counting
methodology which shows the high prevalence of bGH genotype LL among cattle, while
few animals had the LV variant and there was no VV variant was found. A Chi-Square
test analysis of studied population of Pakistani cattle for bGH gene indicated that the
studied population was in Hardy Weinberg equilibrium. Among cattle two alleles A and
B in two genotypic forms AA and AB were found for κ-CN gene fragment. The
frequency of allele A was found to be significantly higher in cattle whereas AB genotype
was more frequent in Sahiwal cattle. No BB genotype was detected in any of these cattle.
The Chi-Square test revealed that observed frequencies and those expected under Hary-
Weinberg were not significant at (P< 0.05), suggesting that Cholistani and Red Sindhi
population was in equilibrium for -CN locus, while animals of Sahiwal cattle were not
in Hary-Weinberg equilibrium. Analysis of β-LG gene, genotypes AA and BB were
detected and no AB genotype was found in the tested animals of Nili-Ravi buffaloes
while the frequency of AA genotype was higher. In cattle observed genotypes were AB
and BB and the frequency of β-LG allele B was significantly higher in the analyzed
animals of three cattle breeds. However, the important thing is the homozygosity found in
Nili-Ravi buffaloes for bGH and κ-CN genes and this could be due to the loss of genetic
variability among the studied population. Nucleotide sequencing from Nili-Ravi buffaloes
shows maximum homology with Bubalus bubalis and Bos taurus breeds. On comparison
of amplified gene sequence of Nili-Ravi buffaloes with Bubalus bubalis and Bos taurus
breeds show variation at several sites.
Thus results show that the PCR-RFLP analysis is easy, cost effective method which
permits easy characterization of bGH, κ-CN and β-LG genes even in the absence of their
expression. This will facilitates the selective breeding programs aimed at increasing the
frequency of desirable alleles.