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利用PCR-SSCP检测荷斯坦奶牛瓜氨酸血症
魏玉春1, 刘丑生2, 王新庄3
1.西北农林科技大学 动物医学院;2.全国畜牧总站畜禽牧草种质资源保存利用中心;3.河南农业大学 牧医工程学院
摘要:
[目的]建立荷斯坦奶牛瓜氨酸血症的PCR-SSCP检测方法.[方法]根据GenBank公布的荷斯坦奶牛精氨酸琥珀酸合成酶基因(Ass)第5外显子核苷酸序列(登录号:M2619),设计并合成1对特异性引物,应用PCR-SSCP方法检测北京市周边牛场173头荷斯坦奶牛瓜氨酸血症的隐性基因,并利用PCR-RFLP和DNA 序列测定验证检测结果的准确性.[结果]PCR-SSCP检测的173头奶牛中有1头是瓜氨酸血症携带者,与PCR-RFLP和DNA 序列测定结果一致,携带率为0.58%.[结论]建立了荷斯坦奶牛瓜氨酸血症的PCR-SSCP检测方法,该方法简单快捷、准确性高、成本低廉,适合荷斯坦奶牛的大规模检测.
关键词:  荷斯坦奶牛  瓜氨酸血症  PCR-SSCP检测
DOI:
分类号:
基金项目:农业部畜禽牧草品种资源保护项目(农财发[2007]43号);科技部《畜禽种质资源》资助项目(2007DKA21101)
Detecting Citrullinemia in Holstein Calves using PCR-SSCP analysis
Abstract:
【Objective】The study established a method to detect Citrullinemia by polymerase chain reaction-single strand conformation polymorphism analysis.【Method】A pair of primers were designed and synthesized according to nucleotide sequences of the exon 5 for argininosuccinate synthesize gene in Holstein Calves,published in Genbank and under accession number:M2619.Polymerase chain reaction-single strand conformation polymorphism was used to analyze exon 5 for argininosuccinate synthesize of 173 Holstein Calves in Beijing,and the results were verified by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing.【Result】 One of the 173 Holstein Calves was Citrullinemia carrier,and the carrying rate was 0.58%.This result was consistent with that of the polymerase chain reaction restriction fragment length polymorphism and DNA sequencing.【Conclusion】 A method detecting Citrullinemia by polymerase chain reaction single strand conformation polymorphism analysis was established.This method is not only simple and convenient,but also has a high accuracy and low cost,which is more suitable for large scale Citrullinemia investigation.
Key words:  Holstein Calf  Citrullinemia  PCR-SSCP detection