Identifikasi keragaman gen FSH sub-unit beta, gen FSH Reseptor dan gen GH pada sapi bali jantan sebagai penanda kualitas sperma

Abstract

The use of markers linked to genes of interest in breeding programmes is more intensive, thus shiffing from phenotype based towards genotype-based selection, have been obvious for many decades. However, realization of this potential has been limited by the lack of markers. With the advent of DNA-based genetic markers in the late 1970s, the situation changed and researchers could, for the first time, begin to identify large numbers of markers dispersed throughout the genetic materials of any species of interest and use the markers to detect associations with traits of interest, thus allowing marker assited selection (MAS) finally to become a reality. The study was conducted to identify polymorphism of Follicle-Stimulating Hormone Sub-beta Unit Gene (FSH beta-sub unit), Follicle- Stimulating Hormone Receptor (FSH receptor) and Growth Hormon (GH) gene had been explored for Sperm Abnormalities Marker in Bali Cattle. A total of 195 samples for association studied from several AI centre of Indonesia and 301 samples for population studied from cattle development area were collected. The objective of this study was to found genetic marker sperm abnormalities in Bali Cattle using DNA polymorphism tools. This study devided in four parts, the first part was study using PCR-RFLP to identify the polymorphism FSH beta-sub unit, the second study using PCR-SSCP, the third study using PCR-RFLP to identify the polymorphism FSH receptor gene, and the fourth study using the PCR-RFLP to identify GH gene. The result in show that Bali cattle was monomorphic that different from other breeds (FH, Brahman, Simmental and Limousin) that were polymorphic. The highest observed heterozygosity was found in Limousin (0.318) and the highest expected heterozygosity was found in Simmental (0.420). Chisquare value was significantly in FH, Brahman and Simmental. The highest incidence of sperm abnormalities were found in Simmental, Limousin and Brahman, while the lowest was found in FH and Bali. In the secondly study, amplification FSH beta-sub unit gene using PCR-SSCP produced allele A and B, also AA, AB and BB genotypes, but in Bali cattle only allele B and BB genotype were found. The highest of observed and expected herezygosity were found in Simmental (0.372), (0.420) respectively. Chi-square values were significant in Brahman, FH and Limousin. The amplification of FSH receptor in the thirdly study using PCR-RFLP with restriction enzym AluI found polymorphism for all breeds. The highest observed heterozygosity was found in Brahman (0.356) and highest expected heterozygosity was found in Simmental (0.420). The chi-square values were significant in all breeds. PCR RFLP with restriction enzym MspI were used to amplify GH gene in fourth study. Results in gene GH|MspI analysis show that only B allele and BB genotype were found in Bali cattle, these indicate that the gene is monomorphic. The diversity of allele frequencies and genotype GH gene indicate that the frequency of allele A was the highest in Simmental (0.750) and lowest in Bali cattle (0.000), allele B was the highest in Bali cattle ii (1,000) and lowest in Simmental (0.250). The highest observed heterozygosity value was found in Simmental (0.500) and the lowest in Bali cattle (0.000), Chisquare value was significant only in FH. Result in association sperm abnormalities indicate lower abnormality in AB genotype when compared to BB and AA genotypes.