【摘要】：利用RT-PCR和RACE技術(shù),從珍稀觀賞植物金花茶(Camellia nitidissima)花瓣中克隆得到了一個(gè)番茄紅素ε-環(huán)化酶(lycopeneε-cyclase,LCYE)基因c DNA全長(zhǎng),命名為CnLCYE。堿基序列分析顯示,CnLCYE基因全長(zhǎng)2 149 bp,包含291 bp的5'非翻譯區(qū)(untranslated regions,UTR)、265 bp的3'UTR和1 593 bp編碼530個(gè)氨基酸的開(kāi)放閱讀框。該基因編碼的蛋白質(zhì)含有2個(gè)跨膜結(jié)構(gòu)域,一個(gè)NADB_Rossmann superfamily結(jié)構(gòu)域以及番茄紅素ε-環(huán)化酶結(jié)構(gòu)域(PLN02697)。CnLCYE蛋白二級(jí)結(jié)構(gòu)以無(wú)規(guī)則卷曲為主,其次為α-螺旋,β-折疊所占比例最少。氨基酸序列比對(duì)分析結(jié)果顯示,CnLCYE與茄科、薔薇科等植物L(fēng)CYE蛋白同源性都在70%以上,與普洱茶(Camellia sinensis var.assamica)LCYE同源性最高。根據(jù)該CnLCYE全長(zhǎng)序列設(shè)計(jì)帶有KpnⅠ和Bam HⅠ位點(diǎn)的全長(zhǎng)擴(kuò)增引物擴(kuò)增基因全長(zhǎng),用KpnⅠ和Bam HⅠ雙酶切后與表達(dá)載體p CAMBIA1300連接,成功構(gòu)建了CnLCYE基因的正義表達(dá)載體,為深入研究CnLCYE基因的功能及其對(duì)花色的調(diào)控作用提供了幫助。
[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 偽-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，

本文編號(hào)：2503124