铁皮石斛RCA2基因克隆与生物信息学分析

发布时间：2019-01-19 12:00

【摘要】：核酮糖1,5-二磷酸羧化酶/加氧酶是光合碳同化作用的关键酶,它对调节铁皮石斛Rubisco活性和光合速率具有直接的作用,对其进行基因等方面的研究,会对改变植物光合速率打下良好基础。该研究以一年生铁皮石斛叶片为材料,采用RT-PCR和RACE技术成功克隆了Rubisco活化酶(RCA)基因,并对其进行生物信息学分析。结果表明:RCA基因全长1 730 bp,命名为RCA2(Gen Bank登录号KT205842),其中5'-UTR 81 bp、3'-UTR 326 bp,开放阅读框1 323 bp,编码440个氨基酸,分子质量为48.53 k Da,等电点为6.19,包含P-loop NTPase超家族基因结构。氨基酸多重序列比对发现RCA2核苷酸序列与蝴蝶兰的相似性高达87%。RCA2编码蛋白为亲水性蛋白;亚细胞定位于叶绿体基质;蛋白质二级结构分析,α螺旋占30.68%,延伸链占25.45%,不规则折叠占43.86%。RCA2编码蛋白质的功能预测发现,RCA2在中间代谢起到了一个非常重要的角色。该研究发现对铁皮石斛光合作用关键酶Rubisco的分析可为其光合作用特性的发掘提供理论基础,并为提高温室大棚栽培效率提供理论依据。
[Abstract]:Ribonose 5-diphosphate carboxylase / oxygenase is the key enzyme of photosynthetic carbon assimilation. It plays a direct role in regulating Rubisco activity and photosynthetic rate of Dendrobium candidum. It will lay a good foundation for changing the photosynthetic rate of plants. In this study, the Rubisco activase (RCA) gene was cloned from the annual leaves of Dendrobium candidum by RT-PCR and RACE techniques, and its bioinformatics analysis was carried out. The results showed that the whole length of RCA gene was 1 730 bp, named RCA2 (Gen Bank accession number KT205842), in which 5'-UTR 81 bp,3'-UTR 326 bp, open reading frame 1 323 bp, encodes 440 amino acids with molecular weight of 48.53 k Da,. The isoelectric point is 6.19, which contains the gene structure of P-loop NTPase superfamily. Amino acid multiple sequence alignment showed that the similarity between RCA2 nucleotide sequence and Phalaenopsis was as high as that of 87%.RCA2 coding protein was hydrophilic, and subcell was located in chloroplast matrix. In protein secondary structure analysis, 伪 helix accounted for 30.68%, extension chain 25.45%, irregular folding accounted for the function prediction of protein encoded by 43.86%.RCA2, and RCA2 played a very important role in intermediate metabolism. It was found that the analysis of Rubisco, a key enzyme in photosynthesis of Dendrobium candidum, could provide a theoretical basis for exploring its photosynthesis characteristics and improving the efficiency of greenhouse cultivation.
【作者单位】：郑州师范学院生物工程研究所;河南农业大学生命科学学院;
【基金】：河南省教育厅重点科研项目(14B180036) 郑州市科技发展计划项目(20150448) 郑州市重大科技专项(141PZDZX038)~~
【分类号】：S567.239

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