金花茶CnLCYE基因cDNA克隆及表达载体构建
发布时间：2019-06-20 09:44
【摘要】：利用RT-PCR和RACE技术,从珍稀观赏植物金花茶(Camellia nitidissima)花瓣中克隆得到了一个番茄红素ε-环化酶(lycopeneε-cyclase,LCYE)基因c DNA全长,命名为CnLCYE。碱基序列分析显示,CnLCYE基因全长2 149 bp,包含291 bp的5'非翻译区(untranslated regions,UTR)、265 bp的3'UTR和1 593 bp编码530个氨基酸的开放阅读框。该基因编码的蛋白质含有2个跨膜结构域,一个NADB_Rossmann superfamily结构域以及番茄红素ε-环化酶结构域(PLN02697)。CnLCYE蛋白二级结构以无规则卷曲为主,其次为α-螺旋,β-折叠所占比例最少。氨基酸序列比对分析结果显示,CnLCYE与茄科、蔷薇科等植物LCYE蛋白同源性都在70%以上,与普洱茶(Camellia sinensis var.assamica)LCYE同源性最高。根据该CnLCYE全长序列设计带有KpnⅠ和Bam HⅠ位点的全长扩增引物扩增基因全长,用KpnⅠ和Bam HⅠ双酶切后与表达载体p CAMBIA1300连接,成功构建了CnLCYE基因的正义表达载体,为深入研究CnLCYE基因的功能及其对花色的调控作用提供了帮助。
[Abstract]:A lycopene e-cyclase (lycopene-cyclase,LCYE) gene c DNA was cloned from the petals of (Camellia nitidissima), a rare ornamental plant, by RT-PCR and RACE, named CnLCYE.. Base sequence analysis showed that the full length of CnLCYE gene was 2149 bp, which contained CnLCYE 5 'untranslated region (untranslated regions,UTR), 265 bp 3'UTR and 1 593 bp open reading frame encoding 530 amino acids. The protein encoded by this gene contains two transmembrane domains, one NADB_Rossmann superfamily domain and lycopene 蔚-cyclase domain (PLN02697). The secondary structure of CnLCyE protein is mainly irregular curl, followed by 伪-helix, and 尾-folding accounts for the least. The results of amino acid sequence alignment analysis showed that CnLCYE had more than 70% homology with LCYE protein of Solanaceae, Rosaceae and other plants, and had the highest homology with (Camellia sinensis var.assamica) LCYE of Pu'er tea. According to the full-length sequence of CnLCYE, the full-length amplification primers with Kpn 鈪，
本文编号：2503124
[Abstract]:A lycopene e-cyclase (lycopene-cyclase,LCYE) gene c DNA was cloned from the petals of (Camellia nitidissima), a rare ornamental plant, by RT-PCR and RACE, named CnLCYE.. Base sequence analysis showed that the full length of CnLCYE gene was 2149 bp, which contained CnLCYE 5 'untranslated region (untranslated regions,UTR), 265 bp 3'UTR and 1 593 bp open reading frame encoding 530 amino acids. The protein encoded by this gene contains two transmembrane domains, one NADB_Rossmann superfamily domain and lycopene 蔚-cyclase domain (PLN02697). The secondary structure of CnLCyE protein is mainly irregular curl, followed by 伪-helix, and 尾-folding accounts for the least. The results of amino acid sequence alignment analysis showed that CnLCYE had more than 70% homology with LCYE protein of Solanaceae, Rosaceae and other plants, and had the highest homology with (Camellia sinensis var.assamica) LCYE of Pu'er tea. According to the full-length sequence of CnLCYE, the full-length amplification primers with Kpn 鈪，
本文编号：2503124
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