Open AccessIdentification of restriction enzyme in the FSHR gene of indonesian local cattle
Author(s) -
Peni Wahyu Prihandini,
A Primasari,
Muhammad Luthfi,
Dicky Pamungkas,
Aprilianna Putri Zahara Nafsina Luvita Sari,
T B Dina,
Dyah Maharani
Publication year - 2021
Publication title -
iop conference series. earth and environmental science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.179
H-Index - 26
eISSN - 1755-1307
pISSN - 1755-1315
DOI - 10.1088/1755-1315/888/1/012024
Subject(s) - restriction enzyme , restriction fragment length polymorphism , restriction site , genotyping , biology , genetics , single nucleotide polymorphism , genbank , allele , snp , microbiology and biotechnology , genotype , gene
The restriction enzyme is important for genotyping using the PCR-RFLP technique. Therefore, this study aims to identify the restriction enzyme mapping in the partial sequence of the follicle-stimulating hormone receptor (FSHR) gene in Indonesian local cattle. A total of 29 samples sized 306 bp, were aligned with Genbank sequence acc no. NC_032660, resulting three polymorphic sites, namely g.193G>C, g.227T>C, and g.275A>C. Furthermore, the restriction mapping analysis using the NEBcutter program V2.0 showed that no enzyme recognized the SNP g.275A>C, while the SNP g.193G>C and g.227T>C were identified by the AluI and MscI enzymes, respectively. The AluI enzyme cuts at two positions (193 bp and 243 bp) in the G allele sample producing three fragments namely 50 bp, 63 bp, and 193 bp, meanwhile, in the C allele, the AluI cuts only in position 243 bp, hence, the fragment products are 63 bp and 243 bp. In contrast, the MscI enzyme was only recognized in the T allele, producing fragments sized 77 bp and 229 bp but failed to identify the restriction site along with the PCR products in the C allele. Based on the results, the SNPs (g.193G>C and g.227T>C) and restriction enzymes ( AluI and MscI ) are applicable for genotyping local Indonesian cattle using the PCR-RFLP technique in future studies.