苦荞漆酶基因的克隆与生物信息学分析  

Molecular Cloning and Bioinformatics Analyzing of Laccase in Fagopyrum tataricum

作　　者：杨晓琳 段迎 蔡苏云 贺润丽 尹桂芳[2] 王艳青[2] 卢文洁[2] 孙道旺[2] 王莉花[2] Yang Xiaolin;Duan Ying;Cai Suyun;He Runli;Yin Guifang;Wang Yanqing;Lu Wenjie;Sun Daowang;Wang Lihua(College of Traditional Chinese Medicine and Food Engineering,Shanxi University of Traditional Chinese Medicine,Taiyuan 030619,Shanxi,China;Biotechnology and Germplasm Resources Institute,Yunnan Academy of Agricultural Sciences/Yunnan Provincial Key Laboratory of Agricultural Biotechnology/Key Laboratory of Southwestern Crop Gene Resources and Germplasm Innovation,Ministry of Agriculture and Rural Affairs,Kunming 650221,Yunnan,China)

机构地区：[1]山西中医药大学中药与食品工程学院,山西太原030619 [2]云南省农业科学院生物技术与种质资源研究所/云南省农业生物技术重点实验室/农业农村部西南作物基因资源与种质创制重点实验室,云南昆明650221

出　　处：《作物杂志》2022年第3期73-79,共7页Crops

基　　金：国家自然科学基金地区科学基金项目(31460379);财政部和农业农村部:国家现代农业产业技术体系(CARS-07-C-2);山西省重点研发计划项目(201803D221012-6);山西省自然科学研究面上项目(20210302123231);校级科技创新能力培育计划(2020PY-YC-16)。

摘　　要：利用RT-PCR技术从苦荞中克隆出2条漆酶(laccase)基因,运用生物信息学技术对2个基因序列进行分析,预测结构域、二级和三级蛋白结构,进行蛋白同源比对及进化树分析。结果表明,FtLAC-1基因序列开放阅读框为1695bp,编码564个氨基酸,FtLAC-2基因序列开放阅读框为1707bp,编码568个氨基酸。FtLAC-1和FtLAC-2蛋白预测分子量分别为61.41和62.47kDa,理论等电点分别为9.45和9.41,为亲水性蛋白,2个基因序列相似性较低,氨基酸序列同源性不高。qRT-PCR分析出其在苦荞不同组织器官存在差异性表达。结论为进一步探索FtLAC-1和FtLAC-2在苦荞薄果壳形成中的功能提供理论基础。In this experiment,to study the gene function of laccase protein in the development of tartary buckwheat,two laccase genes from tartary buckwheat were cloned using RT-PCR technology.Bioinformatics techniques were used to predict the domain,secondary and tertiary protein structure,and to perform the protein homology alignment and phylogenetic evolutionary tree analysis.The results showed that the open reading frame of the FtLAC-1 gene sequence was 1695bp,coding for 564 amino acids,and the open reading frame of the FtLAC-2 gene sequence was 1707bp,coding for 568 amino acids.The predicted molecular weights of FtLAC-1 and FtLAC-2 proteins were 61.41 and 62.47kDa and their theoretical isoelectric points were 9.45 and 9.41,respectively.They were hydrophilic proteins.The sequence similarity of the two genes was low and the amino acid sequence homology was not high.qRT-PCR analysis of its differential expression in different tissues and organs of tartary buckwheat.This study provided a theoretical basis for further exploring the functions of the FtLAC-1 and FtLAC-2 genes in thin buckwheat husk formation.

关 键 词：苦荞 漆酶 基因克隆 生物信息学 表达分析 

分 类 号：S517[农业科学—作物学]