Exploration of Polymorphism Analysis of CD18 Gene with PCR-RFLP Technique in Friesian Holstein Crossbreed Dairy Cows in Batu, East Java, Indonesia
Exploration of Polymorphism Analysis of CD18 Gene with PCR-RFLP Technique in Friesian Holstein Crossbreed Dairy Cows in Batu, East Java, Indonesia
Tiara Widyaputri*, Aldryan Cristianto Pratama, Dian Vidiastuti, Yudit Oktanella
ABSTRACT
This research aims to identify the presence of BLAD carrier alleles and determine whether the polymorphism that appears in the CD18 gene by the PCR-RFLP method is related to the BLAD carrier allele in Friesian Holstein crossbreed dairy cows. A total of 60 samples of cow’s whole blood were collected and extracted and DNA isolated. DNA amplification was carried out using primer Forward (BLAD_F) 5’-TCAACGTGACCTTCCGGAGG-3’ and Reverse (BLAD_R) 5’-CCCAGATTCTTGACGTTGAC-3’. Amplification of the CD18 gene segment on chromosome 1 exon 2 with a target length of 106bp has been successfully carried out with a success rate of 100%. RFLP analysis of NCOI cutting restriction enzymes shows that there are two bands at 66bp and 40bp. This shows that the normal allele frequency is one and the mutant allele is 0 with a heterozygous genotype frequency of 0% in the population. According to the result, the polymorphism that appears in the CD18 gene by PCR-RFLP method in all samples shows the substitution mutation type G base to C transversion and is not related to the BLAD mutation.
Keywords | Dairy cows, CD18 gene, Restriction enzyme, PCR-RFLP, Friesian Holstein crossbreed
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April 2022
Vol. 10, Iss. 4, Pages 712-954
