Genetics Department, Faculty of Agriculture, Fayoum University, Fayoum, 63514, Egypt.
Agricultural Research Center, Animal Production Research Institute, Giza, 12618, Egypt.
J Assist Reprod Genet. 2017 Dec;34(12):1683-1690. doi: 10.1007/s10815-017-1007-2. Epub 2017 Jul 31.
This study explores polymorphisms in the growth differentiation factor 9 (GDF9) gene (exon 1) with respect to fertility in Egyptian sheep.
Blood samples were collected, and genomic DNA was extracted from 24 Saidi and 13 Ossimi ewes. A 710 bp portion of the GDF9 gene, was amplified using specific primers, and the sequence was analyzed to clarify the phylogenetic relationship of Egyptian breed sheep. In addition, the PCR-RFLP method using Pst1 or Msp1 restriction enzymes was used to mask polymorphisms of partial exon 1 of GDF9 gene to establish molecular markers for twinning.
The lambing rate percentage and litter size showed significant difference between ewes, which produce single and twin lamb for each breed individually, whereas the coefficient of variation of the Saidi breed is greater than that of the Ossimi breed. The results suggested that the GDF9 gene shared a similarity in sequence compared to six accession numbers of Ovis aries found in GenBank. Molecular phylogenetic analyses were performed based on nucleotide sequences in order to examine the position of the Egyptian breeds among many other sheep breeds. The results indicate that accession number AF078545 of O. aries is closely related with Saidi and Ossimi ewes that produce single or twin lamb using the unweighted pair group method with arithmetic mean (UPGMA) analysis. Results showed that Msp1 enzyme digestion revealed polymorphic restriction pattern consisting of one band with 710 bp for ewes producing single lamb and two bands with 710 and 600 bp for ewes producing twin lamb in Saidi sheep breed.
Sequence analysis and diversity of polymorphisms in the GDF9 gene (exon 1) have a novel base substitution (A-T) for detection of Fec mutations that serve as a molecular marker for twinning.
本研究探讨了生长分化因子 9（GDF9）基因（exon 1）多态性与埃及绵羊繁殖力的关系。
采集 24 只赛迪羊和 13 只奥西姆羊的血液样本，提取基因组 DNA。使用特异性引物扩增 GDF9 基因的 710 bp 部分，分析序列以阐明埃及绵羊品种的系统发育关系。此外，使用 Pst1 或 Msp1 限制酶的 PCR-RFLP 方法来掩盖 GDF9 基因部分外显子 1 的多态性，以建立与双胎相关的分子标记。
两种绵羊的产羔率和产羔数存在显著差异，每个品种的母羊分别产单胎或双胎，而赛迪品种的变异系数大于奥西姆品种。结果表明，GDF9 基因与 GenBank 中发现的 6 个绵羊 Ovis aries 序列具有相似性。基于核苷酸序列进行分子系统发育分析，以检验埃及绵羊品种在许多其他绵羊品种中的位置。结果表明，绵羊 O. aries 的 AF078545 登录号与产单胎或双胎的赛迪和奥西姆母羊密切相关，使用非加权配对组平均法（UPGMA）分析。结果表明，Msp1 酶消化显示出多态性限制图谱，710 bp 为单胎产羔母羊，710 和 600 bp 为双胎产羔母羊。
GDF9 基因（exon 1）序列分析和多态性多样性具有新的碱基替换（A-T），可用于检测 Fec 突变，作为双胎的分子标记。
