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A novel SNP marker of ovalbumin gene in association with duck hatchability

机译:卵清蛋白基因的新型SNP标记与鸭孵化率相关

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Our previous transcriptome analysis using a cDNA microarray identified differentially-expressed transcripts in Tsaiya ducks (Anas platyrhynchos); we concluded that the ovalbumin gene might be involved in duck hatchability. In the present study, associations of single nucleotide polymorphism (SNP) genotypes of the duck ovalbumin gene with hatchability were investigated. To confirm the cDNA microarray analysis, real-time polymerase chain reaction (PCR) and Western blot analysis were used to validate ovalbumin gene expression. The messenger RNA and protein expression of the ovalbumin gene were higher (P < 0.05) in the low-hatchability group (1.00 +/- 0.19; 30.36 +/- 3.51 arbitrary units) than in high-hatchability counterparts (0.56 +/- 0.07; 8.53 +/- 2.97 arbitrary units), consistent with the previous cDNA microarray analysis. The PCR products (506 base pairs) of ovalbumin gene amplified by the primer pair of TovaF and TovaR from the genomic DNA templates of 10 ducks were sequenced and a g.385 C>T SNP site in the 506-base pair sequence of the ovalbumin gene identified. Genotyping of SNP of 187 ducks was then carried out by PCR restriction fragment length polymorphism and minisequencing methods. Based on SNP genotypes of the duck ovalbumin gene, there were three types: CC, TT, and CT. Birds with the CC and TT genotypes had higher hatchability (79.59 +/- 3.40, 76.35 +/- 1.77) (P < 0.05) than those with a CT genotype (65.77 +/- 2.07). In conclusion, the ovalbumin gene was an important candidate gene that can be used for marker-assisted selection to increase hatchability in Tsaiya ducks. (C) 2013 Elsevier Inc. All rights reserved.
机译:我们之前使用cDNA微阵列进行的转录组分析确定了Tsaiya鸭(Anas platyrhynchos)中差异表达的转录本;我们得出的结论是卵清蛋白基因可能与鸭的孵化率有关。在本研究中,研究了鸭卵清蛋白基因的单核苷酸多态性(SNP)基因型与孵化率的关联。为了确认cDNA微阵列分析,实时聚合酶链反应(PCR)和蛋白质印迹分析用于验证卵清蛋白基因表达。低孵化率组(1.00 +/- 0.19; 30.36 +/- 3.51任意单位)卵清蛋白基因的信使RNA和蛋白质表达高于高孵化率的对应物(0.56 +/- 0.07)(P <0.05) ; 8.53 +/- 2.97任意单位),与以前的cDNA微阵列分析一致。用TovaF和TovaR引物对从10只鸭子的基因组DNA模板中扩增出卵白蛋白基因的PCR产物(506个碱基对),并在卵白蛋白506个碱基对的序列中建立了一个g.385 C> T SNP位点。基因鉴定。然后通过PCR限制性片段长度多态性和微测序方法对187只鸭子的SNP进行基因分型。根据鸭卵清蛋白基因的SNP基因型,可分为CC,TT和CT三种类型。具有CC和TT基因型的鸟的孵化率(79.59 +/- 3.40,76.35 +/- 1.77)(P <0.05)比具有CT基因型的鸟(65.77 +/- 2.07)高。总之,卵清蛋白基因是重要的候选基因,可用于标记辅助选择以增加Tsaiya鸭的孵化率。 (C)2013 Elsevier Inc.保留所有权利。

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