基于聚合酶链反应的伊朗荷斯坦牛群体中牛淋巴细胞抗原DRB3.2的限制性片段长度多态性基因分型

PCR based RFLP genotyping of bovine lymphocyte antigen DRB3.2 in Iranian Holstein population.

作者信息

Pashmi Morteza, Qanbari Saber, Ghorashi Seyed Ali, Salehi Abdolreza

机构信息

Animal Genetics Division, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.

出版信息

Pak J Biol Sci. 2007 Feb 1;10(3):383-7. doi: 10.3923/pjbs.2007.383.387.

DOI:10.3923/pjbs.2007.383.387

PMID:19069505

Abstract

Major Histocompatibility Complex (MHC) class II locus DRB3 was investigated by PCR based restriction fragment length polymorphism (PCR-RFLP) assay. A total of 262 Holstein cows participating in the national recording system were sampled from 10 herds. A two-step polymerase chain reaction was carried out in order to amplify a 284 base-pair fragment of exon 2 of the target gene. Second PCR products were treated with three restriction endonucleas enzymes RsaI, BstYI and HaeIII. Digested fragments were analyzed by polyacrylamid gel electrophoresis. Twenty-eight BoLA-DRB3 alleles were identified. Identified alleles are: BoLA-DRB3.2 *3, *6, *7, *8, *9, *10, *11, *12, *13, *14, *15, *16,20, *21, *22, *23, *24, *25, *26, *27, *28, *32, *36, *37, *40, *51, *iaa and ibb. The BoLA-DRB3.240 allele that was observed in this study has not been reported previously. The calculated frequencies were as follows: 2.29, 1.34, 0.19, 14.5, 0.38, 3.05, 12.21, 1.34, 2.29, 1.34, 2.48, 9.16, 0.95, 0.77, 6.68, 9.16, 17.94, 1.15, 0.57, 1.15, 0.95, 0.57, 0.38, 1.91, 0.38, 5.73, 0.19 and 0.95% respectively. The six most frequently observed alleles (BoLA-DRB3.2 *8, *11, *16, *22, *23 and *24) accounted for 69.65% of the alleles in these 10 herds. The results of this study confirm the allelic distribution of six most frequent alleles in Holstein population's worldwide.

摘要

采用基于聚合酶链反应的限制性片段长度多态性（PCR-RFLP）分析方法，对主要组织相容性复合体（MHC）II类基因座DRB3进行了研究。从10个牛群中选取了262头参与国家记录系统的荷斯坦奶牛作为样本。进行了两步聚合酶链反应，以扩增目标基因外显子2的一个284碱基对的片段。第二步PCR产物用三种限制性内切酶RsaI、BstYI和HaeIII进行处理。消化后的片段通过聚丙烯酰胺凝胶电泳进行分析。共鉴定出28个BoLA-DRB3等位基因。鉴定出的等位基因有：BoLA-DRB3.2 *3、*6、*7、*8、*9、*10、*11、*12、*13、*14、*15、*16、20、*21、*22、*23、*24、*25、*26、*27、*28、*32、*36、*37、*40、51、iaa和ibb。本研究中观察到的BoLA-DRB3.240等位基因此前未见报道。计算得到的频率如下：分别为2.29%、1.34%、0.19%、14.5%、0.38%、3.05%、12.21%、1.34%、2.29%、1.34%、2.48%、9.16%、0.95%、0.77%、6.68%、9.16%、17.94%、1.15%、0.57%、1.15%、0.95%、0.57%、0.38%、1.91%、0.38%、5.73%、0.19%和0.95%。观察到的六个最常见等位基因（BoLA-DRB3.2 *8、*11、*16、*22、23和24）占这10个牛群中所有等位基因的69.65%。本研究结果证实了全球荷斯坦牛群体中六个最常见等位基因的等位基因分布情况。