{"title":"用PCR-RFLP方法分析巴阳鸭胰岛素样生长因子-1基因多态性","authors":"T. Rafian, Y. Yurnalis","doi":"10.31186/jspi.id.18.3.162-168","DOIUrl":null,"url":null,"abstract":"This study aims to determine the polymorphism of the IGF-1 gene in Bayang ducks using the PCR-RFLP method. This study used 120 blood samples of Bayang ducks in Kampung Binuang Dalam, Padang City, West Sumatra Province. Bayang duck blood samples were taken through the brachial vein for ± 1 mL. DNA extraction was taken from the blood using the Genomic DNA Purification Kit (Promega) Protocol. DNA was amplified using a pair of primers F: 5'- CCA GGA ATA TCT TTG GAA GCT GT-3 and R: 5'- TGC TAC GTT ACC AGC CTT GA -3 'which produced a 433 bp fragment of exon 3 IGF-1 gene, and F: 5'- CTG GAG CAG GCA GGA AAA TT - 3 'and R: 5'- TCC AGG GAC AGT GAC TCA AC -3' which produced 801 bp fragments of exon 4 IGF-1 gene. The amplification product was restricted by the DdeI enzyme, which recognized the C↓TNAG slashing site (N = G, A, T, C) for the exon three and MnII regions, which recognized the CAC↓GTG cutting site for the exon four regions. The cutting of the IGF-1 exon three gene product in Bayang ducks using DdeI as a restriction enzyme only recognized the C↓TNAG cutting site. The cutting results visualized with 2% agarose gel showed two genotypes: homozygous (+ / +) of 41 samples and the heterozygote (+/-) genotype of 60 samples. The cutting of the IGF-1 exon four gene product in Shadow ducks using MnII as a restriction enzyme only recognized the CAC↓GTG cutting site. The cutting results visualized with 2% agarose gel showed three types of genotypes, namely four samples homozygous (+/+), 67 samples heterozygous (+/-), and 49 samples homozygous (-/-). Based on the results of the study, it can be concluded that the IGF-1 gene in Bayang ducks is polymorphic, with the frequency of the IGF-1|DdeI genotype being in a hardy-weinberg imbalance, and the IGF-1|MnII genotype in a hardy-weinberg equilibrium.","PeriodicalId":33136,"journal":{"name":"Jurnal Sain Peternakan Indonesia","volume":"28 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2023-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Polymorphism of Insulin-Like Growth Factor-1 (IGF-1) Gene on Bayang Ducks Using PCR-RFLP Method\",\"authors\":\"T. Rafian, Y. Yurnalis\",\"doi\":\"10.31186/jspi.id.18.3.162-168\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"This study aims to determine the polymorphism of the IGF-1 gene in Bayang ducks using the PCR-RFLP method. This study used 120 blood samples of Bayang ducks in Kampung Binuang Dalam, Padang City, West Sumatra Province. Bayang duck blood samples were taken through the brachial vein for ± 1 mL. DNA extraction was taken from the blood using the Genomic DNA Purification Kit (Promega) Protocol. DNA was amplified using a pair of primers F: 5'- CCA GGA ATA TCT TTG GAA GCT GT-3 and R: 5'- TGC TAC GTT ACC AGC CTT GA -3 'which produced a 433 bp fragment of exon 3 IGF-1 gene, and F: 5'- CTG GAG CAG GCA GGA AAA TT - 3 'and R: 5'- TCC AGG GAC AGT GAC TCA AC -3' which produced 801 bp fragments of exon 4 IGF-1 gene. The amplification product was restricted by the DdeI enzyme, which recognized the C↓TNAG slashing site (N = G, A, T, C) for the exon three and MnII regions, which recognized the CAC↓GTG cutting site for the exon four regions. The cutting of the IGF-1 exon three gene product in Bayang ducks using DdeI as a restriction enzyme only recognized the C↓TNAG cutting site. The cutting results visualized with 2% agarose gel showed two genotypes: homozygous (+ / +) of 41 samples and the heterozygote (+/-) genotype of 60 samples. The cutting of the IGF-1 exon four gene product in Shadow ducks using MnII as a restriction enzyme only recognized the CAC↓GTG cutting site. The cutting results visualized with 2% agarose gel showed three types of genotypes, namely four samples homozygous (+/+), 67 samples heterozygous (+/-), and 49 samples homozygous (-/-). Based on the results of the study, it can be concluded that the IGF-1 gene in Bayang ducks is polymorphic, with the frequency of the IGF-1|DdeI genotype being in a hardy-weinberg imbalance, and the IGF-1|MnII genotype in a hardy-weinberg equilibrium.\",\"PeriodicalId\":33136,\"journal\":{\"name\":\"Jurnal Sain Peternakan Indonesia\",\"volume\":\"28 1\",\"pages\":\"0\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-09-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Jurnal Sain Peternakan Indonesia\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.31186/jspi.id.18.3.162-168\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Jurnal Sain Peternakan Indonesia","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.31186/jspi.id.18.3.162-168","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
本研究旨在利用PCR-RFLP方法测定巴阳鸭IGF-1基因多态性。本研究使用了西苏门答腊省巴东市Kampung Binuang Dalam的120只巴阳鸭的血液样本。巴阳鸭经肱静脉取血±1ml,采用基因组DNA纯化试剂盒(Promega)方案提取血样中的DNA。用一对引物扩增DNA, F: 5′- CCA GGA ATA TCT TTG GAA GCT GT-3和R: 5′- TGC TAC GTT ACC AGC CTT GA -3′产生了433 bp的IGF-1基因外显子片段,F: 5′- CTG GAG CAG GCA GGA AAA TT -3′和R: 5′- TCC AGG GAC AGT GAC TCA AC -3′产生了801 bp的IGF-1基因外显子片段。扩增产物受到DdeI酶的限制,DdeI酶识别外显子3和MnII区的C↓TNAG切割位点(N = G, A, T, C), DdeI酶识别外显子4区的CAC↓GTG切割位点。利用DdeI作为限制性内切酶切割巴阳鸭IGF-1外显子3基因产物,只识别C↓TNAG切割位点。用2%琼脂糖凝胶显像,结果显示41份样品为纯合子(+/ +)型,60份样品为杂合子(+/-)型。使用MnII作为限制性内切酶切割影子鸭IGF-1外显子4基因产物,只识别CAC↓GTG切割位点。2%琼脂糖凝胶可视化切割结果显示3种基因型,即纯合子(+/+)4份,杂合子(+/-)67份,纯合子(-/-)49份。本研究结果表明,巴阳鸭IGF-1基因具有多态性,IGF-1|DdeI基因型频率处于hardy-weinberg失衡,IGF-1|MnII基因型频率处于hardy-weinberg平衡。
Polymorphism of Insulin-Like Growth Factor-1 (IGF-1) Gene on Bayang Ducks Using PCR-RFLP Method
This study aims to determine the polymorphism of the IGF-1 gene in Bayang ducks using the PCR-RFLP method. This study used 120 blood samples of Bayang ducks in Kampung Binuang Dalam, Padang City, West Sumatra Province. Bayang duck blood samples were taken through the brachial vein for ± 1 mL. DNA extraction was taken from the blood using the Genomic DNA Purification Kit (Promega) Protocol. DNA was amplified using a pair of primers F: 5'- CCA GGA ATA TCT TTG GAA GCT GT-3 and R: 5'- TGC TAC GTT ACC AGC CTT GA -3 'which produced a 433 bp fragment of exon 3 IGF-1 gene, and F: 5'- CTG GAG CAG GCA GGA AAA TT - 3 'and R: 5'- TCC AGG GAC AGT GAC TCA AC -3' which produced 801 bp fragments of exon 4 IGF-1 gene. The amplification product was restricted by the DdeI enzyme, which recognized the C↓TNAG slashing site (N = G, A, T, C) for the exon three and MnII regions, which recognized the CAC↓GTG cutting site for the exon four regions. The cutting of the IGF-1 exon three gene product in Bayang ducks using DdeI as a restriction enzyme only recognized the C↓TNAG cutting site. The cutting results visualized with 2% agarose gel showed two genotypes: homozygous (+ / +) of 41 samples and the heterozygote (+/-) genotype of 60 samples. The cutting of the IGF-1 exon four gene product in Shadow ducks using MnII as a restriction enzyme only recognized the CAC↓GTG cutting site. The cutting results visualized with 2% agarose gel showed three types of genotypes, namely four samples homozygous (+/+), 67 samples heterozygous (+/-), and 49 samples homozygous (-/-). Based on the results of the study, it can be concluded that the IGF-1 gene in Bayang ducks is polymorphic, with the frequency of the IGF-1|DdeI genotype being in a hardy-weinberg imbalance, and the IGF-1|MnII genotype in a hardy-weinberg equilibrium.