目的 以藏红花悬浮细胞为研究对象，克隆藏红花酸糖基转移酶（crocetin glycosyltransferase）UGTCs4基因，并对其进行生物信息学和表达分析。方法 采用同源克隆法和5’RACE（rapid amplification of cDNA ends）技术克隆UGTCs4基因的全长cDNA序列。运用生物信息学方法对该基因进行分析，预测其编码蛋白的结构与功能，并通过半定量RT-PCR（semi-quantitative reverse transcription and polymerase Chainreaction）方法检测不同诱导物条件下的基因表达情况。结果 UGTCs4基因的cDNA全长为1380 bp，编码459个氨基酸。分子进化树分析，该糖基转移酶基因与已知的藏红花酸糖基转移酶基因UGTCs2具有相同的糖基化藏红花酸的功能。RT-PCR的结果显示，UGTCs4基因能够响应吲哚乙酸（IAA）、脱落酸（ABA）、赤霉素（GA）、过氧化氢（H₂O₂）、茉莉酸甲酯（MeJA）多种处理，这些处理均能促使其进行转录。结论 首次从藏红花悬浮细胞中分离并报道了UGTCs4的cDNA克隆，并证实UGTCs4对不同诱导子的响应情况，为进一步研究藏红花素的生物合成及表达调控研究奠定了基础。

Objective With the aim to obtain crocetin glycosyltransferase UGTCs4 in cultured saffron suspension cell, and carry out its bioinformatics and expression mode analysis.Methods A homologous cloning strategy and 5' RACE methods were adopted, the full-length cDNA sequence of a crocetin glycosyltransferase, designated UGTCs4 (GeneBank number:KX398932), was obtained. The characteristics of physiochemical properties, structure and function of the deduced UGTCs4 protein were determined using a series of bioinformatics tools. Semi-quantitative PCR was used for gene expression analysis. Results The results showed that the full length cDNA of UGTCs4 was 1 380 bp in length and encoding a 459 amino acid; UGTCs4 had high identities (83.2%) with UGTCs2 protein from saffron; UTGTCs4 had the same evolutionary tree as UGTCs2. UGTCs4 transcripts were constitutively expressed in the leaves, stems, and roots. UGTCs4 gene could respond to multiple treatments of indoleacetic acid (IAA), abscisic acid (ABA), gibberellin (GA), hydrogen peroxide (H₂O₂), and methyl jasmonate (MJA), which promoted its transcription. Conclusion cDNA of crocetin glycosyltransferase was cloned from suspension cells for the first time and the response of UGTCs4 to different inducers was confirmed. Molecular characterization of UGTCs4 will be useful for further functional determination of the gene involving in the crocin biosynthesis and expression regulation.

R282.12

国家自然科学基金项目（C020409）；新疆自治区高技术项目（201211107）