| 刘倩倩，李彩霞，赵 静，谷从璟，张晓东.滇龙胆香叶醇合酶基因的克隆与表达分析[J].广东农业科学,2017,44(9):48-55 |
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滇龙胆香叶醇合酶基因的克隆与表达分析 |
| Cloning and expression analysis of geraniol synthase gene in Gentiana rigescens |
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| DOI：10.16768/j.issn.1004-874X.2017.09.008 |
| 中文关键词: 滇龙胆 香叶醇合酶 基因克隆 表达分析 |
| 英文关键词: Gentiana rigescens geraniol synthase gene cloning expression analysis |
| 基金项目:云南省大学生创新项目（201511390005，201511390006）；云南省教育厅重点项目（2015Z171） |
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| 中文摘要: |
| 对滇龙胆香叶醇合酶基因GrGES 进行克隆和表达分析，为研究其在香叶醇和龙胆苦苷生物合
成中的作用奠定基础。根据滇龙胆转录组GrGES 序列，采用RT-PCR 技术从滇龙胆叶片克隆该基因，获得
GrGES 序列（GenBank 登录号为KJ917168）。序列分析结果表明，GrGES 基因长1 767 bp，编码588 氨基
酸；GrGES 蛋白相对分子质量为67.84 ku，理论pI 为5.56。该蛋白定位于叶绿体，包含叶绿体转运肽，为亲
水不稳定蛋白，主要由α- 螺旋（62.41%）和环（39.57%）构成。该蛋白具有其他GES 蛋白的活性位点、
萜类合酶N 端结构域（IPR001906，81～273）和萜类合酶结构域（IPR005630，263～588）。GrGES 蛋白与
长春花CrGES 蛋白亲缘关系最近。原核表达结果表明，GrGES 基因在大肠杆菌中表达的重组蛋白相对分子
质量约为78.22 ku，与预期的大小一致。组织特异性表达分析结果表明，GrGES 基因主要在叶中表达。 |
| 英文摘要: |
| GrGES（ geraniol synthase） gene was cloned and its expression characters were examined in Gentiana
rigescens，which would lay foundations for studies of function of this gene in geraniol and gentiopicroside biosynthesis.
GrGES was cloned from leaves of G. rigescens by RT-PCR according to the GrGES sequence of transcriptome of G.
rigescens. As a result，GrGES with its GenBank accession number KJ917168 was obtained. Sequence analysis
showed that GrGES gene had a length of 1 767 bp coding for 588 amino acids. Its relative molecular weight was
67.84 ku with the theoretical isoelectric point of 5.56. GrGES may localize in chloroplast，and it was a hydropholic
unstable protein composed of mainly α-helix（ 62.41%） and loop（ 39.57%）. The active sites in other GESs，
N-terminal domain of terpene synthase（ IPR001906，81-273），domain of terpene synthas（ IPR005630，
263-588） were all existed in GrGES. GrGES protein was close to CrGES in Catharanthus roseus. The results of
prokaryotic expression of GrGES gene in E. coli indicated that the recombinant protein was approximately 78.22
ku，which was consistent with the presumptive size. The tissue-specific expression results showed that GrGES gene
was mainly expressed in leaf. |
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