摘要
【目的】分离半番鸭羽色相关基因,探索半番鸭羽色基因表达调控的分子机制。【方法】以半番鸭白羽皮肤和黑羽皮肤分别作为试验组(tester)和驱动组(driver),利用双链特异性核酸酶(duplexspecificnuclease,DSN)介导的均一化消减杂交方法,构建半番鸭羽色相关基因消减cDNA文库,同时采用实时荧光定量PCR方法对文库质量进行验证。【结果】①从文库中随机选取144个阳性克隆进行测序分析,最终共获得64条有效序列,平均长度为1 031 bp;经BLAST比对发现,21条具有同源序列,且同源性平均为92.8%;43条未找到匹配序列,推测可能为羽色相关新基因;②Geneontology功能分析表明,已知基因参与信号转导、细胞结构、物质转运、细胞凋亡、细胞与机体防御、转录与表达调控等诸多生物学过程,并且与色素形成和转运存在不同程度的相关性。【结论】经实时荧光定量PCR鉴定后,所建文库质量良好,能够有效地富集白羽皮肤特异表达基因。
[Objective] The aim of this study was to isolate plumage color related-genes in mule duck and explore its molecular mechanism. [Method] A normalized subtraetive library of plumage color related-genes of mule duck was constructed. White plumage skin and black plumage skin were adopted respectively as the tester and driver, and a duplex-specific nuclease （DSN） normalized subtractive hybridization method was used to construct the library, and real-time PCR was employed to identify the quality of the library. [Result] Results showed that 64 genes with the average size of 1 031 bp were obtained by selecting and sequencing 144 positive clones randomly. Nucleotide BLAST homological analysis incidated that 21 genes had similarities to known genes which enjoy 92.8% homology, and 43 genes were not matched which presumably may be new genes related to plumage color. Geneontology displayed that these known genes were involved in many biological processes such as signal transduction, cell structure, material transport, apoptosis, cell and organism defense, transcription and expression regulation, and had different correlations with formation and transshipment of pigment. [Conclusion] It was determined that the library had good quality and could enrich the genes of the white plumage via real-time PCR.
出处
《中国农业科学》
CAS
CSCD
北大核心
2012年第22期4688-4696,共9页
Scientia Agricultura Sinica
基金
福建省自然基金项目(2010J01107)
福建省种业创新与产业化工程项目(2011FJZY-9-02)
现代农业产业技术体系建设专项(nycytx-45-06)
