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技术说明:通过聚合酶链反应-单链构象多态性同时鉴定山羊CSN1S2 A、B、C和E等位基因

Technical note: simultaneous identification of CSN1S2 A, B, C, and E alleles in goats by polymerase chain reaction-single strand conformation polymorphism.

作者信息

Chessa S, Rignanese D, Chiatti F, Radeghieri A, Gigliotti C, Caroli A

机构信息

Dipartimento di Scienze e Tecnologie Veterinarie per Sicurezza Alimentare, Università degli Studi di Milano, Via Trentacoste 2, Milano 20134, Italy.

出版信息

J Dairy Sci. 2008 Mar;91(3):1214-7. doi: 10.3168/jds.2007-0653.

Abstract

Most variability in goat caseins originates from the high number of genetic polymorphisms often affecting the specific protein expression, with strong effects on milk composition traits and technological properties. At least 7 alleles have been found in the goat alpha(S2)-CN gene (CSN1S2). Five of them (CSN1S2A, CSN1S2B, CSN1S2C, CSN1S2E, and CSN1S2F) are widespread in most breeds, whereas the other 2 (CSN1S2D and CSN1S2*0) are rarer alleles. Four different PCR-RFLP tests are needed to detect all of these variants at the DNA level. The objective of this study was to develop and validate a rapid method for typing 4 of the 5 most-common goat CSN1S2 alleles by means of PCR-single strand conformation polymorphism (SSCP). The method was validated by analyzing 37 goat samples at the protein and DNA level, respectively, by milk isoelectrofocusing and PCR-RFLP methods already described. The genotypes obtained using the PCR-SSCP approach were in full agreement with those obtained by the validation analyses. The newly developed PCR-SSCP approach provides an accurate and inexpensive assay highly suitable for genotyping goat CSN1S2.

摘要

山羊酪蛋白的大多数变异源于大量通常影响特定蛋白质表达的基因多态性,这对牛奶成分特征和工艺特性有很大影响。在山羊α(S2)-CN基因(CSN1S2)中已发现至少7个等位基因。其中5个(CSN1S2A、CSN1S2B、CSN1S2C、CSN1S2E和CSN1S2F)在大多数品种中广泛存在,而另外2个(CSN1S2D和CSN1S2*0)是较罕见的等位基因。需要四种不同的PCR-RFLP测试来在DNA水平检测所有这些变体。本研究的目的是开发并验证一种通过PCR-单链构象多态性(SSCP)对5个最常见的山羊CSN1S2等位基因中的4个进行分型的快速方法。通过分别使用已描述的牛奶等电聚焦和PCR-RFLP方法在蛋白质和DNA水平分析37个山羊样本,对该方法进行了验证。使用PCR-SSCP方法获得的基因型与通过验证分析获得的基因型完全一致。新开发的PCR-SSCP方法提供了一种准确且廉价的检测方法,非常适合对山羊CSN1S2进行基因分型。

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