目的 克隆布渣叶查耳酮合酶基因（MpCHS）全长cDNA，并对其在不同部位和不同生长阶段叶片中的表达模式进行分析。方法 以布渣叶叶片总RNA逆转录合成cDNA为模板，根据其转录组数据设计特异引物序列，PCR扩增MpCHS基因全长cDNA，经质粒连接、转化、扩培，挑选阳性克隆测序、分析并构建原核表达载体。同时，采用实时荧光定量PCR（RT-qPCR）对MpCHS基因的表达模式进行分析。结果 成功克隆得到MpCHS基因全长cDNA（GenBank：KY472608）。生物信息学分析表明其开放阅读框为1 176 bp，编码含391个氨基酸的蛋白，其相对分子质量为42 700，理论等电点6.11，具有CHS家族蛋白3个保守的功能活性位点（165 C、304 H和337 N）和特征多肽标签序列RLMMYQQGCFAGGTVLR和GVLFGFGPGL。系统进化树分析发现MpCHS与可可、陆地棉等木本植物的亲缘关系比较近。RT-qPCR结果表明，MpCHS基因在不同部位均有表达，在叶片中的表达量随着生长过程逐步降低。结论 首次克隆得到MpCHS基因，并分析了MpCHS基因在布渣叶不同部位和不同生长阶段叶片中的表达模式，为MpCHS基因的原核表达和功能验证奠定了基础，也为进一步解析布渣叶黄酮类生物合成途径提供了参考。

Objective To clone the full-length cDNA of MpCHS and analyze its expression pattern in different parts of Microcos paniculata and different growth periods of M. paniculata leaves. Methods Based on the transcriptome data of M. paniculata, we designed specific primers for MpCHS gene. The full-length cDNA of MpCHS was amplified by PCR and the positive clones were then sequenced, analyzed, and constructed prokaryotic expression vector. The bioinformatics analysis of MpCHS was also performed. Meanwhile, the mRNA expression of MpCHS was detected using real-time quantitative PCR. Results The relative molecular mass was 42 700, and its theoretical isoelectric point was 6.11, with three conserved functional active sites (165 C, 304 H, and 337 N) of the CHS family proteins and the tag sequence of RLMMYQQGCFAGGTVLR and GVLFGFGPGL. Phylogenetic tree analysis showed that MpCHS had close relationship with woody plants such as cocoa and upland cotton. We successfully cloned the full-length cDNA of MpCHS (GenBank:KY472608). It had an ORF of 1 176 bp which encoded a protein of 391 amino acid residues. RT-qPCR results showed that MpCHS was expressed in all parts of M. paniculata and its expression in leaves was gradually decreased along with its development. Conclusion MpCHS is cloned from M. paniculate for the first time, and the gene expression pattern of MpCHS in different parts of M. paniculata and different growth periods of M. paniculata leaves was analyzed. This study facilitates the further purification and functional validation of MpCHS protein and provides reference for further analysis of flavonoids biosynthesis pathway in M. paniculata.

国家自然科学基金项目（31300273）；广东省科技计划项目（2015A030302082）