PCR-RFLP genotyping protocol for chicken Mx gene G/A polymorphism associated with the S631N mutation

Sironi, Laura; Ramelli, P.; Williams, J. L.; Mariani, Paola

doi:10.4238/vol9-2gmr811

Cited by 14 publications

(15 citation statements)

References 13 publications

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“…The PCR-RFLP method is a widely used method for genotyping of gene (Hao et al, 2007;Jauk et al, 2003;Koeken et al, 2002;Sironi et al, 2010;Wang et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Genotyping of flavin-containing mono-oxygenase 3 (FMO3) gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mismatch amplification mutation assay (MAMA-PCR) in chickens

Hamid¹,

Wang²,

Zhao

2012

Afr. J. Biotechnol.

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Fishy smelling off-flavour of chicken meat is caused by a mutation (FMO3 c.985A>T) in exon 7 of flavincontaining mono-oxygenase 3 (FMO3) gene. Comparative analysis of the causative mutation in FMO3 gene in different chickens using two different methods was reported. Polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) method was based on general primers, whereas another employed method was the mismatch amplification mutation assay (MAMA) primers. Seventy (22.88%) of the 306 samples were genotyped using PCR-RFLP, 194 (63.40%) of the 306 samples were genotyped using MAMA-PCR and 42 (13.72%) of the 306 samples were genotyped by both of PCR-RFLP and MAMA-PCR and genotyping data were validated by DNA sequencing. The results show that the genotyping accuracy for PCR-RFLP was 90.48% in AA, 85.42% in AT and 95.35% in TT and 100% in AA, 98% in AT and 96% in TT for MAMA-PCR. This study suggests that the MAMA-PCR is more effective, accurate, easy, fast, efficient and reliable than PCR-RFLP.

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“…The PCR-RFLP method is a widely used method for genotyping of gene (Hao et al, 2007;Jauk et al, 2003;Koeken et al, 2002;Sironi et al, 2010;Wang et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

Genotyping of flavin-containing mono-oxygenase 3 (FMO3) gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mismatch amplification mutation assay (MAMA-PCR) in chickens

Hamid¹,

Wang²,

Zhao

2012

Afr. J. Biotechnol.

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“…The DNA fragment was amplified with 25 μL of PCR solution consisting of 12.5 µL of Mastermix PCR (MyTaq HS Red Bioline), 1.25 μL of forwarding primer (5ʹ-GCACTGTCACCTCTTAATAGA-3ʹ), and 1.25 μL of reverse primer (5ʹ-GTATTGGTAGGCTTTGTTGA-3ʹ), 5 μL of DNA samples, and 5 μL of ddH2O. PCR amplification was performed using a thermal cycler PCR machine with a predenaturation condition at 95 °C for 3 min, denaturation at 95 °C for 15 s, annealing at 60 °C for 15 s, extension at 72 °C for 25 s, and post extension at 72 °C for 5 min (Sironi et al 2010). The PCR products were detected through electrophoresis on 2% agarose gel in a submarine electrophoresis system (Mupid-EXU) device.…”

Section: Dna Isolation and Sequencingmentioning

confidence: 99%

Expression of Mx exon-13 SNPs in Kampong-Laying Type (Kamper) chicken crossbreeds of female Lohmann brown-classic and male Pelung

et al. 2020

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Abstract. Afifah D, Lesmana I, Poerwanto SH, Joko T, Mahardhika IWS, Daryono BS. 2020. Expression of Mx exon-13 SNPs in Kampong-Laying Type (Kamper) chicken crossbreeds of female Lohmann brown-classic and male Pelung. Biodiversitas 21: 1483-1487. The Gama Ayam Research Team has implemented marker-assisted selection (MAS) in selective breeding to provide a faster, more accurate, and more reliable selection of chicken. Mx gene expression has a vital role in chicken disease resistance. This research aimed to investigate the expression of Mx exon-13 single nucleotide polymorphisms (SNPs) in the population of female Layer Lohmann Brown-Classic, male Pelung, and its progenies Kampong-Laying Type (Kamper) chicken. The G1892A mutation in Mx exon-13 resulted in a change in the amino acid 631 of Mx. The substitution of serine to asparagine favored the ability of chickens to acquire immunity against viral diseases, including avian influenza. Asparagine (A allele) at position 631 is specific to Mx+ (resistant), whereas serine (G allele) is specific to Mx− (susceptible). DNA was amplified using the forward primer 5ʹ-GCACTGTCACCTCTTAATAGA-3ʹ and the reverse primer 5ʹ-GTATTGGTAGGCTTTGTTGA-3ʹ and then sequenced using the Sanger sequencing method. Four SNPs were obtained through Mx sequence alignment. They consisted of four substitutions (A20734T, C20737T, A20766G, and A20893G) with one haplotype. Mx exon-13 SNPs were detected in Pelung and Kamper. Therefore, Kamper chicken inherited the disease resistance gene of Pelung and could be a strong candidate for parental generation in the further selective breeding program.

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“…Primer yang digunakan untuk mengamplifikasi gen Mx berdasarkan Sironi et al (2010) adalah primer forward (5'-GCA CTG TCA CCT CTT AAT AGA-3') dan primer reverse (5'GTA TTG GTA GGC TTT GTT GA-3'). Primer tersebut mampu mengamplifikasi gen Mx sepanjang 299 pb yaitu dari basa ke 20 670-20 968 (exon 13).…”

Section: Amplifikasi Pcrunclassified

Keragaman Gen Myxovirus (Mx) pada Ayam Kampung Terseleksi Polymorphisms of Myxovirus Gene in Selected Kampung Chicken

Permatasari

Sumantri

Darwati

et al. 2015

JIPTHP

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The Mx gene is wellknown as marker genetic for chicken resistance of virus infection. The aim of this study was to analyze the mx gene polymorphisms in selected kampung chicken. A total of 25 heads of kampung chicken, which had been selected for ND virus resistant, were identified by PCR-RFLP method.The Mx gene used Hpy8I restriction enzyme where digested at GTN|NAC nucleotides. There were two alleles, A and G with three genotypes AA, AG dan GG. The result showed that 69.2% of kampung chicken were resistant toward the virus infection. This population was in Hardy-Weinberg equilibrium (x 2 < x 2 0.05

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PCR-RFLP genotyping protocol for chicken Mx gene G/A polymorphism associated with the S631N mutation

Cited by 14 publications

References 13 publications

Genotyping of flavin-containing mono-oxygenase 3 (FMO3) gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mismatch amplification mutation assay (MAMA-PCR) in chickens

Genotyping of flavin-containing mono-oxygenase 3 (FMO3) gene by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and mismatch amplification mutation assay (MAMA-PCR) in chickens

Expression of Mx exon-13 SNPs in Kampong-Laying Type (Kamper) chicken crossbreeds of female Lohmann brown-classic and male Pelung

Keragaman Gen Myxovirus (Mx) pada Ayam Kampung Terseleksi Polymorphisms of Myxovirus Gene in Selected Kampung Chicken

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