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首页> 外文期刊>Journal of Molecular Genetics >Polymorphism of β-Lactoglobulin Gene in Iranian Sheep Breeds Using PCR-RFLP
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Polymorphism of β-Lactoglobulin Gene in Iranian Sheep Breeds Using PCR-RFLP

机译：PCR-RFLP技术检测伊朗绵羊品种β-乳球蛋白基因多态性

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β-lactoglobulin is the major milk whey protein in ruminants and its coding gene located on ovine chromosome 3. This protein produces in mammary glands during pregnancy and lactation stages. Studies have shown that the protein is polymorphic in many breeds of sheep. This is the result of single base pair substitution in β-lactoglobulin gene that also rises to RsaI restriction Fragment Length Polymorphism (RFLP). Blood samples were supplied from 142 animals of Iranian sheep breeds (Ghezel, Afshary, Moghani, Makoii and Arkharmerino). Genomic DNA was extracted from 100 μL blood sample according to Boom method modified by Shaikhayev. Gel monitoring and spectrophotometeric methods were used for determination of DNA quality and quantity. BLg 5 and 3 primers amplified a 452 bp fragment from exon II of ovine β-lactoglobulin gene. Products of amplification were recognized by electrophoresis on 1% agarose gel stained with ethidium bromide. RsaI enzyme was used for restriction of PCR products. Digestion products were separated by electrophoresis on 8% non-denaturant polyacrylamide gel and visualized after staining with ethidium bromide on UV gel documentation. PopGen 32 software (ver. 1.31) was used to estimating the frequency of allele, genotype and Hardy-Weinberg equilibrium. Frequency of A-allele in Ghezel, Afshary, Moghani, Makoii and Arkharmerino sheep was 56, 34, 36, 53 and 48%, respectively. The populations were in Hardy-Weinberg equilibrium except of Afshary breed.
机译：β-乳球蛋白是反刍动物中的主要乳清蛋白，其编码基因位于绵羊3号染色体上。该蛋白在妊娠和哺乳期的乳腺中产生。研究表明，这种蛋白质在许多绵羊品种中都是多态的。这是β-乳球蛋白基因中单碱基对取代的结果，该基因也上升为RsaI限制性片段长度多态性（RFLP）。从142个伊朗绵羊品种（Ghezel，Afshary，Moghani，Makoii和Arkharmerino）的动物中采集血液样本。根据Shaikhayev修改的Boom方法，从100μL血液样本中提取基因组DNA。凝胶监测和分光光度法用于测定DNA的质量和数量。 BLg 5和3引物从绵羊β-乳球蛋白基因外显子II扩增了452 bp片段。通过在溴化乙锭染色的1％琼脂糖凝胶上电泳识别扩增产物。 RsaI酶用于限制PCR产物。消化产物通过在8％非变性聚丙烯酰胺凝胶上的电泳进行分离，并在UV凝胶文献中用溴化乙锭染色后可视化。使用PopGen 32软件（1.31版）估计等位基因的频率，基因型和Hardy-Weinberg平衡。 Ghezel，Afshary，Moghani，Makoii和Arkharmerino绵羊的A等位基因频率分别为56％，34％，36％，53％和48％。除Afshary品种外，其他种群处于Hardy-Weinberg平衡状态。

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