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[摘要]
目的 对杯萼海桑Sonneratia alba萜类化合物生物合成途径的关键酶法呢基焦磷酸合成酶(Farnesyl pyrophosphate synthase,FPS)基因的编码cDNA序列进行克隆,为研究杯萼海桑萜类化合物生物合成与基因调控奠定基础。方法 结合杯萼海桑根的转录组注释,根据编码区序列设计引物,通过PCR方法克隆杯萼海桑FPS(SaFPS)基因的编码区cDNA。结果 PCR扩增了一个长1 029 bp的基因片段,该片段编码由342个氨基酸组成的SaFPS。同源性比对结果显示基因编码蛋白与甘草的FPS具有氨基酸一致性达86%。其具有异戊烯基转移酶的2个典型保守功能域。进化树分析结果显示,与甘草Glycyrrhiza uralensis、苜蓿Medicago truncatula 、羽扇豆Lupinus albu 具有较近的亲缘关系。实时荧光定量PCR结果显示SaFPS基因在花中表达量较高,果实、茎、叶中表达量相对较低。结论 首次从红树林植物杯萼海桑中克隆FPS基因获得其编码区序列,SaFPS基因具有组织表达特异性。本研究为分析基因表达特性及其在生物合成中的功能奠定基础。
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[Abstract]
Objective To clone the full length cDNA encoding Farnesyl pyrophosphate synthase (FPS) which played an important role in terpenoid biosynthesis pathway in Sonneratia alba and to provide the basis for the further studies on biosynthesis and gene regulation of terpenoid. Methods According to the annotation of root transcriptome in S. alba, primers were designed and cDNA of S. alba Farnesyl pyrophosphate synthase (SaFPS) gene was cloned from S. alba by PCR. Results The complete coding sequence of SaFPS gene was 1 029 bp and encoded a protein of 342 amino acids. The deduced SaFPS amino acid sequence exhibited 86% identity to the FPS of Glycyrrhiza uralensis. The predicted SaFPS shared two conserved functional domains. Phylogenetic analysis on the amino acid sequence of SaFPS with those of other plants showed that SaFPS was closely related to G. uralensis, Medicago truncatula, and Lupinus albus. It showed that SaFPS was highly expressed in flowers while lowly expressed in fruits, stems, and leaves by real-time PCR analysis. Conclusion The SaFPS gene is cloned from a mangrove tree S. alba for the first time and the expression of SaFPS is tissue specific. This research lays a foundation for studying the gene expression pattern and regulating the functions of SaFPS in terpenoid biosynthesis.
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