摘要
【目的】克隆鉴定杧果CHS基因,分析其功能,明确杧果不同组织器官中CHS基因表达情况。【方法】以海南种植的6个杧果种的叶片总DNA和c DNA为模板,进行同源性PCR扩增,然后克隆到p MD18-T载体上,进行测序;通过生物学软件预测分析CHS基因编码的蛋白质;运用半定量PCR分析CHS基因表达情况。【结果】测序得到6个杧果种CHS基因c DNA全长为1 173 bp,CHS基因含有2个外显子和1个内含子。基于c DNA序列的NJ系统发育树显示,其与普通杧果(Mangifera indica)CHS1基因同源性最高。CHS1基因半定量表达显示杧果不同器官均表达CHS1基因,但在叶片和花中的表达量较高。【结论】克隆了杧果查尔酮合成酶1(CHS1)基因,半定量分析了杧果不同器官中CHS1基因的表达差异,为下一步研究提高CHS1基因的表达量奠定了基础。
[Objective]The aims of this study was to clone the chalcone synthase （CHS） gene and analyze its function, and reveal its expression in different tissues of mango. [Methods ] Total DNA and RNA from Mango were used as template to conduct PCR amplification and a target fragment in the amplification was cloned to vector pMD18-T easily and then sequenced. The structure of the protein （CHS） was analyzed by bioinformatics. The expression analysis was finished by Semi-quantitative PCR. [Results] Sequence analysis showed that the eDNA of CHS was 1 173 bp in length and contained two exons and one intron. The phylogenetic analyze based on the eDNA sequences showed that CHS shares high homology with the CHS1 from Mangifera indica. Semi-quantitative analysis showed that the CHS1 gene can be detected in all Mango tissues, but the gene expression levels in leaves and flowers were higher than those in other tissues. [Conclusion] In this study, the CHS1 gene was cloned and analyzed for the expression in different tissues, which has laid the foundation for increasing the expression of CHS1.
出处
《果树学报》
CAS
CSCD
北大核心
2015年第6期1077-1084,I0003,共9页
Journal of Fruit Science
基金
公益性行业(农业)科研专项项目(201203092-2)
物种资源保护项目(12RZZY-07)
国家自然科学基金(31460455)
