茶树叶绿体靶向铁氧化还原蛋白nadp +氧化还原酶基因的克隆与表达分析Huangjinya

Xiuxiu Zhao, Yangen Fan, Yueyue Tian, Hanyue Wang, Lixia Zhang, Min Li
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引用次数: 0

摘要

叶绿体靶铁氧化还原酶是一种位于高等植物光合作用电子传递链末端的亲水黄素蛋白。它是连接光合作用电子传输和叶绿体氧化还原代谢的枢纽。为了探究LFNR基因与‘黄金牙’‘Shuchazao’(CK)光控黄变的关系,我们分别以‘黄金牙’‘Shuchazao’(CK)的一芽两叶为材料克隆了CsLFNR基因,得到了相似度为100%的序列,命名为CsLFNR1.1 (GenBank登录号:MT311318)。通过生物信息学分析,我们知道其cDNA基因长度为1 095 bp,编码364个氨基酸,蛋白质分子量为40.623KDa,理论等电点为8.86。cslfnr1.1是一种碱性蛋白,无跨膜结构和信号肽,具有一个叶绿体转运肽(cTP),二级结构包含23.35%的α螺旋、5.49%的β角、28.85%的延伸链和42.31%的不规则卷曲,位于叶绿体中。通过Nucleotide BLAST、Protein BLAST和DNAMAN软件,我们发现CsLFNR1.1及其翻译的氨基酸序列与NCBI‘Shuchazao’基因CsLFNR1 (XM_028233617.1)和Protein (XP_028089418.1)有4个碱基和3个氨基酸的差异,相似度分别达到99.63%和99.18%。CsLFNR1.1和CsLFNR1在理化性质和二级结构上差异不大,三级结构预测模板一致。此外,它们具有相同的保守结构域和活性位点。我们以不同遮荫的‘黄金崖’叶片为材料进行qRT-PCR,结果表明CsLFNR1.1基因的表达对光强有响应,其表达量随着光强的增加而增加。本研究为进一步探索CsLFNR1.1基因在‘黄金牙’新芽新叶光调控过程中的作用提供了理论基础和科学依据。
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Cloning and Expression Analysis of Chloroplast-targeted Ferredoxin-NADP+ Oxidoreductase Gene of Tea Plant (Camellia sinensis) cv.Huangjinya
The chloroplast-targeted ferredoxin-NADP+ oxidoreductase (LFNR) is a hydrophilic flavin protein located at the end of the photosynthesis electron delivery chain in higher plants. It’s a hub for linking photosynthesis electronic transportation with chloroplast redox metabolism. In order to explore the relationship between LFNR gene and yellowing of ‘Huangjinya’controlled by light, we took separately one bud two leaves of ‘Huangjinya’ ‘Shuchazao’(CK) as the material to clone CsLFNR gene and then we obtained the sequence with a similarity of 100%, named CsLFNR1.1 (GenBank accession no. MT311318). Throw bioinformatics analysis, we know its cDNA gene length is 1 095 bp, coding 364 amino acids, The protein molecular weight is 40.623KDa, the theoretical isoelectric point is 8.86.CsLFNR1.1 is a alkaline protein without transmembrane structure and signal peptide, it has a chloroplast transport peptide (cTP), the secondary structure contains 23.35% alpha helix, 5.49% beta-corner, 28.85% extension chain and 42.31% irregular curling, It locates in the chloroplast. Through the Nucleotide BLAST, Protein BLAST and DNAMAN software, we found that CsLFNR1.1 and its translated amino acid sequences had 4 bases and 3 amino acid differences with NCBI ‘Shuchazao’ Gene CsLFNR1 (XM_028233617.1) and Protein (XP_028089418.1), the similarity reached 99.63% and 99.18%. CsLFNR1.1 and CsLFNR1 have small differences in the physical and chemical properties and secondary structure, the tertiary structure prediction template of them is consistent. In addition, they have the same conserved domain and active site. We took different shade ‘Huangjinya’ blades as the material for qRT-PCR, the result showed that the expression of CsLFNR1.1 gene response to the light intensity, its expression increased with the increase of light intensity. This study provides a theoretical basis and scientific basis for further exploring the role of CsLFNR1.1 gene in the light regulation process of the new shoots and leaves of ‘Huangjinya’.
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