茶樹(shù)Copine家族基因CsBON3的克隆與表達(dá)分析
發(fā)布時(shí)間:2018-05-04 02:51
本文選題:茶樹(shù) + Copine家族。 參考:《茶葉科學(xué)》2017年06期
【摘要】:Copine蛋白是一類(lèi)包含2個(gè)C2(N端)和1個(gè)vWA(C端)保守域的Ca~(2+)依賴蛋白或磷脂結(jié)合蛋白,在胞內(nèi)信號(hào)轉(zhuǎn)導(dǎo)中發(fā)揮重要作用;谛蛄邢嗨菩苑治,從茶樹(shù)轉(zhuǎn)錄組數(shù)據(jù)庫(kù)中篩選出1條與Copine家族基因高度同源的EST序列。經(jīng)測(cè)序驗(yàn)證該序列包含1 746 bp的完整ORF,編碼581個(gè)氨基酸。同源比對(duì)顯示該基因與擬南芥At BON3序列相似度最高(65%),將其命名為CsBON3(Gen Bank登錄號(hào)為KY435900)。生物信息學(xué)分析顯示,CsBON3蛋白分子量為63.66 k D,理論等電點(diǎn)為5.48;具有Copine家族蛋白特有的保守結(jié)構(gòu)域;屬親水性蛋白,無(wú)信號(hào)肽位點(diǎn),非分泌性蛋白,無(wú)跨膜結(jié)構(gòu)域。表達(dá)分析表明,CsBON3在茶樹(shù)花和根系中表達(dá)量最高,莖干和成熟葉中表達(dá)量最低。低溫(4℃)處理茶樹(shù)1 d后,其表達(dá)被顯著上調(diào);在生長(zhǎng)階段,該基因表達(dá)量高于休眠階段;同時(shí)在接種炭疽菌的茶樹(shù)葉片中,該基因也被快速上調(diào),表明該基因可能與茶樹(shù)低溫、生長(zhǎng)發(fā)育及抗病相關(guān)。
[Abstract]:Copine proteins are a class of Ca~(2 / phospholipid binding proteins that contain two C2N terminal and one vWA(C conserved domain and play an important role in intracellular signal transduction. Based on sequence similarity analysis, a EST sequence with high homology to Copine family genes was screened from tea transcriptional database. The sequence was confirmed to contain 1 746 BP complete ORF encoding 581 amino acids. Homologous alignment showed that the gene had the highest similarity with Arabidopsis thaliana at BON3 sequence, and it was named CsBON3(Gen Bank accession number KY435900. Bioinformatics analysis showed that the molecular weight of CsBON3 protein was 63.66kD, and the theoretical isoelectric point was 5.48. it had the conserved domain of Copine family protein, which belonged to hydrophilic protein, no signal peptide site, non-secretory protein and no transmembrane domain. The expression of CsBON3 was the highest in tea flower and root system, and the lowest in stem and mature leaf. The expression of the gene was significantly up-regulated in tea plants treated at 4 鈩,
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