广藿香香叶醇-10羟化酶基因克隆及在不同栽培种中的表达分析
发布时间：2018-10-05 06:52
【摘要】：目的:克隆香叶醇-10羟化酶(geraniol 10-hydroxylase,G10H)基因,进行生物信息学分析,并在不同广藿香栽培种中分析不同时期的茎、叶的表达情况。方法:搜索广藿香转录组数据库,获得G10H基因全长序列,并设计全长引物进行PCR验证;利用生物信息学软件对PcG10H1基因进行生物信息学分析,实时定量PCR法对其进行了时空表达分析。结果:将获得的广藿香香叶醇-10羟化酶基因命名为PcG10H1(Gen Bank登录号为KF926077),该基因包含一个完整的长1 533 bp的开放阅读编码框,编码510个氨基酸。分析了PcG10H1基因编码蛋白的理化特性。分析发现其有1个跨膜区,无信号肽;PcG10H基因编码的氨基酸序列与胡黄连G10H基因编码的氨基酸序列最为接近;PcG10H1在老叶和老茎中表达量高,在老叶中表达量最高;从不同栽培种来看,PcG10H1在石牌广藿香和高要广藿香中表达模式相似,在海南广藿香与印尼广藿香中表达相似。结论:成功克隆了广藿香PcG10H1的全长序列且进行了序列分析,并对其在不同栽培种的表达模式进行了探讨,为进一步阐明广藿香萜类代谢途径奠定基础。
[Abstract]:Objective: to clone the gene of geraniol 10-hydroxylase G10H and analyze its bioinformatics, and to analyze the expression of stem and leaf in different stages of cultivation of Pogostemon patchouli. Methods: the full-length sequence of G10H gene was obtained by searching the database of transcription group of Pogostemon patchouli, and the full-length primer was designed for PCR verification, and the bioinformatics analysis of PcG10H1 gene was carried out by bioinformatics software. Real-time quantitative PCR method was used to analyze its temporal and spatial expression. Results: the obtained gene was named PcG10H1 (Gen Bank accession number (KF926077). The gene contained a complete 1 533 bp open reading coding frame encoding 510 amino acids. The physicochemical properties of the protein encoded by PcG10H1 gene were analyzed. It was found that there was a transmembrane region of PcG10H gene, and the amino acid sequence of PcG10H gene was the closest to that of Coptis chinensis G10H gene. The expression of PcG10H1 was higher in the old leaves and stems, and the highest in the old leaves. The expression patterns of PcG10H1 were similar in Shipai patchouli and Gaoyao patchouli, and were similar in Hainan patchouli and Indonesian patchouli. Conclusion: the full-length sequence of patchouli PcG10H1 was cloned and sequenced, and its expression patterns in different cultivated species were discussed, which laid a foundation for further elucidating the metabolic pathway of terpenes of patchouli.
【作者单位】： 广东食品药品职业学院;
【基金】：广东省科技计划项目(2015A040404029) 广东食品药品职业学院自然科学研究项目(2015YZ007) 广东省医学科研基金项目(A2016621)
【分类号】：S567.239
[Abstract]:Objective: to clone the gene of geraniol 10-hydroxylase G10H and analyze its bioinformatics, and to analyze the expression of stem and leaf in different stages of cultivation of Pogostemon patchouli. Methods: the full-length sequence of G10H gene was obtained by searching the database of transcription group of Pogostemon patchouli, and the full-length primer was designed for PCR verification, and the bioinformatics analysis of PcG10H1 gene was carried out by bioinformatics software. Real-time quantitative PCR method was used to analyze its temporal and spatial expression. Results: the obtained gene was named PcG10H1 (Gen Bank accession number (KF926077). The gene contained a complete 1 533 bp open reading coding frame encoding 510 amino acids. The physicochemical properties of the protein encoded by PcG10H1 gene were analyzed. It was found that there was a transmembrane region of PcG10H gene, and the amino acid sequence of PcG10H gene was the closest to that of Coptis chinensis G10H gene. The expression of PcG10H1 was higher in the old leaves and stems, and the highest in the old leaves. The expression patterns of PcG10H1 were similar in Shipai patchouli and Gaoyao patchouli, and were similar in Hainan patchouli and Indonesian patchouli. Conclusion: the full-length sequence of patchouli PcG10H1 was cloned and sequenced, and its expression patterns in different cultivated species were discussed, which laid a foundation for further elucidating the metabolic pathway of terpenes of patchouli.
【作者单位】： 广东食品药品职业学院;
【基金】：广东省科技计划项目(2015A040404029) 广东食品药品职业学院自然科学研究项目(2015YZ007) 广东省医学科研基金项目(A2016621)
【分类号】：S567.239
【参考文献】
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