本文選題：鐵皮石斛 + ATP-binding��； 參考：《中草藥》2017年15期

【摘要】：目的克隆鐵皮石斛Dendrobium officinale ABC(ATP-binding cassette)轉(zhuǎn)運(yùn)蛋白F家族基因并進(jìn)行生物信息學(xué)分析。方法利用RACE從鐵皮石斛葉片cDNA中分離ABC基因,并進(jìn)行編碼蛋白相對(duì)分子質(zhì)量、等電點(diǎn)、結(jié)構(gòu)域、信號(hào)肽、跨膜域及亞細(xì)胞定位等生物信息學(xué)分析;采用DNASTAR和MEGA6進(jìn)行氨基酸多序列比對(duì)和分子進(jìn)化分析;借助實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)基因組織表達(dá)模式。結(jié)果從鐵皮石斛中分離到2個(gè)F家族ABC轉(zhuǎn)運(yùn)蛋白基因DoABCF1和DoABCF2(Gen Bank注冊(cè)號(hào)KU160474和KU160475),全長(zhǎng)為2 104 bp和2 193 bp,各編碼1條由600和659個(gè)氨基酸組成的肽鏈,相對(duì)分子質(zhì)量67 030和74 140,等電點(diǎn)6.20和5.71;DoABCF1和DoABCF2蛋白均包含2個(gè)保守的ABC結(jié)構(gòu)域(分別為74~314、385~600和65~323、392~607)和多個(gè)基元;2個(gè)蛋白不含信號(hào)肽或跨膜域,預(yù)測(cè)均定位在葉綠體。2個(gè)基因與植物F家族ABC轉(zhuǎn)運(yùn)蛋白基因相似性高達(dá)80%以上,與玉米和水稻等單子葉植物F家族ABC轉(zhuǎn)運(yùn)蛋白基因親緣關(guān)系較近。DoABCF1和DoABCF2基因轉(zhuǎn)錄本在石斛3個(gè)器官中差異表達(dá)且均在葉中高度表達(dá),莖和根中相對(duì)表達(dá)量差異不顯著;以莖為校正樣本,前者在根中相對(duì)表達(dá)量為莖中的1.74倍,后者在葉中表達(dá)量為莖中的3.44倍。結(jié)論成功獲得DoABCF1和DoABCF2基因全長(zhǎng)cDNA,二者在鐵皮石斛葉中的高表達(dá)特征暗示其可能在鐵皮石斛生長(zhǎng)發(fā)育過程中起一定作用。
[Abstract]:Objective to clone Dendrobium officinale ABC (ATP-binding cassette) transporter F gene and analyze it by bioinformatics. Methods ABC gene was isolated from the cDNA of Dendrobium candidum leaves by race, and the relative molecular weight, isoelectric point, domain, signal peptide, transmembrane domain and subcellular localization of the encoded protein were analyzed by bioinformatics. DNASTAR and MEGA6 were used for amino acid multi-sequence alignment and molecular evolution analysis, and real-time fluorescence quantitative PCR was used to detect the expression pattern of gene tissue. Results two F family ABC transporter genes, DoABCF1 and DoABCF2 (GenBank registration numbers KU160474 and KU160475), were isolated from Dendrobium candidum with a length of 2 104 BP and 2 193 BP, each encoding a peptide chain consisting of 600 and 659 amino acids. Relative molecular weight of 67 030 and 74 140, isoelectric points 6.20 and 5.71 of DoABCF1 and DoABCF2 proteins both contain two conserved ABC domains (74 / 314385 / 600 and 65 / 323392C / 607, respectively) and multiple motifs, and two proteins do not contain signal peptides or transmembrane domains. The similarity between the two genes and the ABC transporter genes of the F family of plants was more than 80%. The relationship between ABC transporter genes of F family in maize and rice was close to that of DoABCF1 and DoABCF2 gene transcripts in three organs of Dendrobium and were highly expressed in leaves, but there was no significant difference in relative expression between stem and root. The relative expression of the former in the root was 1.74 times that in the stem, and the latter in the leaf was 3.44 times of that in the stem. Conclusion the full length cDNAs of DoABCF1 and DoABCF2 genes were successfully obtained. Their high expression in the leaves of Dendrobium candidum suggests that they may play a role in the growth and development of Dendrobium candidum.
【作者單位】： 陜西中醫(yī)藥大學(xué)藥學(xué)院陜西省中藥基礎(chǔ)與新藥研究重點(diǎn)實(shí)驗(yàn)室;陜西中醫(yī)藥大學(xué)附屬醫(yī)院第一臨床醫(yī)學(xué)院;中國(guó)醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)學(xué)院藥用植物研究所;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31101608) 陜西省自然科學(xué)基金項(xiàng)目(2017JM8030) 陜西省青年科技新星項(xiàng)目(2012KJXX-44)
【分類號(hào)】：Q943.2;S567.239

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本文編號(hào)：2080588