Türkiye'de yetiştirilen bazı sığır ırklarında sığır lökosit bağlanma yetmezliğinin (BLAD) restriksiyon parçacık uzunluk polimorfizmi (RFLP) ile belirlenmesi
Özet
Detection of Bovine Leukocyte Adhesion Deficiency (BLAD) in Some Turkish Cattle Breeds with Restriction Fragment Length Polymorphism (RFLP) The purpose of this work was to study BLAD allele is present in some cattle breeds in Turkey. It was also intended to establish the molecular genetic test used for this reason in Turkey and to provide infrastructure for the routine use of BLAD screening in the calves which are breeding candidate stock in Holstein breed. The material of the study consisted of the bulls used in the progeny testing program, carried out by The Cattle Breeders' Association of Turkey and the Ministry of Agriculture. The study was done in Ankara University, Faculty of Veterinary Medicine Department of Genetics, Laboratory of Molecular Genetics. It was also concluded that PCR-RFLP method can be successfully used to screen the young breeding cattle whether they are carrying BLAD allele. The blood samples obtained from 120 Holstein, 20 Brown Swiss, 20 Anatolian Black, 20 Turkish Grey, 20 South Anatolian Red and 20 East Anatolian Red. The DNA isolation was made from the blood samples using the method of phenol-chloroform extraction. The isolated DNA materials were multiplied in PCR using the primer developed by Kriegsemann et all. In order to determine the area of mutation in PCR products, the PCR products were digested by Taq I endonuclease enzyme. The resulting fragments were analysed on 2 % agarose gel for the absence of a Taq I restiriction site. It was found that one male and another female two of the Holstein cattle were heterozygote BLAD carriers. There was no homozygote BLAD animals. BLAD allele was not found in the other breeds used in the study. The mutant BLAD allele frequency in the 120 Holstein cattle calculation was 0.0084. This is the first study reporting the presence of BLAD in Holstein cattle breed in Turkey. Key Words: BLAD, Holstein, PCR, restriction enzyme, RFLP.