Hassanane Mohamed S, Hassan Amal A M, Ahmed Fatma M, El-Komy Esteftah M, Roushdy Khaled M, Hassan Nagwa A
Cell Biology Department, National Research Centre, Egypt.
Animal Production Department, National Research Centre, Egypt.
J Genet Eng Biotechnol. 2018 Jun;16(1):83-88. doi: 10.1016/j.jgeb.2017.11.002. Epub 2017 Nov 22.
Egyptian chickens, representing 2 breeds and 7 strains, were genotyped using the PCR-RFLP and sequencing techniques for detection of a non-synonymous dimorphism (G/A) in exon 14 of chicken Myxovirus resistance () gene. This dimorphic position is responsible for altering protein's antiviral activity. Polymerase Chain reactions were performed using Egyptian chickens DNA and specific primer set to amplify DNA fragments of 299 or 301 bp, containing the dimorphic position. Amplicons were cut with restriction enzyme 81. Genotype and allele frequencies for the resistant allele A and sensitive allele G were calculated in all the tested chickens. Results of PCR-RFLP were confirmed by sequencing. The three genotypes AA, AG, GG at the target nucleotide position in gene were represented in all the studied Egyptian chicken breeds and strains except Baladi strain which showed only one genotype AA. The average allele frequency of the resistant A allele in the tested birds (0.67) was higher than the sensitive G allele average frequency in the same birds (0.33). Appling PCR-RFLP technique in the breeding program can be used to select chickens carrying the A allele with high frequencies. This will help in improving poultry breeding in Egypt by producing infectious disease-resistant chickens.
对代表2个品种和7个品系的埃及鸡进行基因分型，采用聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）和测序技术，检测鸡抗黏液病毒（Mx）基因第14外显子中的非同义双态性（G/A）。这个双态性位点负责改变蛋白质的抗病毒活性。使用埃及鸡的DNA和特异性引物对进行聚合酶链反应，以扩增包含双态性位点的299或301 bp的DNA片段。扩增产物用限制性内切酶HaeIII切割。计算所有受试鸡中抗性等位基因A和敏感等位基因G的基因型和等位基因频率。PCR-RFLP的结果通过测序得到证实。除了仅显示一种基因型AA的巴莱迪品系外，在所研究的所有埃及鸡品种和品系中，Mx基因目标核苷酸位置的三种基因型AA、AG、GG均有出现。受试鸡中抗性A等位基因的平均等位基因频率（0.67）高于同一批鸡中敏感G等位基因的平均频率（0.33）。在育种计划中应用PCR-RFLP技术可用于选择高频携带A等位基因的鸡。这将有助于通过培育抗传染病的鸡来改善埃及的家禽育种。
