【摘要】：目的:克隆香葉醇-10羥化酶(geraniol 10-hydroxylase,G10H)基因,進(jìn)行生物信息學(xué)分析,并在不同廣藿香栽培種中分析不同時(shí)期的莖、葉的表達(dá)情況。方法:搜索廣藿香轉(zhuǎn)錄組數(shù)據(jù)庫(kù),獲得G10H基因全長(zhǎng)序列,并設(shè)計(jì)全長(zhǎng)引物進(jìn)行PCR驗(yàn)證;利用生物信息學(xué)軟件對(duì)PcG10H1基因進(jìn)行生物信息學(xué)分析,實(shí)時(shí)定量PCR法對(duì)其進(jìn)行了時(shí)空表達(dá)分析。結(jié)果:將獲得的廣藿香香葉醇-10羥化酶基因命名為PcG10H1(Gen Bank登錄號(hào)為KF926077),該基因包含一個(gè)完整的長(zhǎng)1 533 bp的開(kāi)放閱讀編碼框,編碼510個(gè)氨基酸。分析了PcG10H1基因編碼蛋白的理化特性。分析發(fā)現(xiàn)其有1個(gè)跨膜區(qū),無(wú)信號(hào)肽;PcG10H基因編碼的氨基酸序列與胡黃連G10H基因編碼的氨基酸序列最為接近;PcG10H1在老葉和老莖中表達(dá)量高,在老葉中表達(dá)量最高;從不同栽培種來(lái)看,PcG10H1在石牌廣藿香和高要廣藿香中表達(dá)模式相似,在海南廣藿香與印尼廣藿香中表達(dá)相似。結(jié)論:成功克隆了廣藿香PcG10H1的全長(zhǎng)序列且進(jìn)行了序列分析,并對(duì)其在不同栽培種的表達(dá)模式進(jìn)行了探討,為進(jìn)一步闡明廣藿香萜類(lèi)代謝途徑奠定基礎(chǔ)。
[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.
【作者單位】： 廣東食品藥品職業(yè)學(xué)院;
【基金】：廣東省科技計(jì)劃項(xiàng)目(2015A040404029) 廣東食品藥品職業(yè)學(xué)院自然科學(xué)研究項(xiàng)目(2015YZ007) 廣東省醫(yī)學(xué)科研基金項(xiàng)目(A2016621)
【分類(lèi)號(hào)】：S567.239

【參考文獻(xiàn)】

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