狭叶柴胡β-amyrin合成酶基因的克隆及其蛋白的生物信息学分析  

Cloning and Bioinformatics Analysis ofβ-amyrin Synthase Gene from Bupleurum scorzonerifolium

作　　者：刘艳玲 向凤宁[2] LIU Yan-ling;XIANG Feng-ning(College of Medicine and Nursing,Dezhou University,Shandong Dezhou 253023,China;Key Laboratory of Plant Cell Engineering and Germplasm Innovation,Ministry of Education,School of Life Sciences,Shandong University,Shandong Qingdao 266237,China)

机构地区：[1]德州学院医药与护理学院,山东德州253023 [2]山东大学生命科学院植物细胞工程与种质创新教育部重点实验室,山东青岛266237

出　　处：《西南农业学报》2020年第11期2426-2431,共6页Southwest China Journal of Agricultural Sciences

基　　金：德州学院博士科研基金(2019xgrc25)。

摘　　要：【目的】本文拟获得β-amyrin合成酶基因的全长序列及其蛋白的生物学信息。【方法】以狭叶柴胡为材料,获得了抗肝炎有效成分齐墩果酸合成关键酶—β-amyrin合成酶基因的cDNA序列,并应用生物信息数据库(哪个数据库)和分析软件(交代分析软件的类型)对该基因的保守序列、蛋白质理化性质、跨膜结构域、亚细胞定位、二级结构、三级结构、磷酸化位点做了分析。【结果】β-amyrin合成酶基因的ORF核苷酸长度2289 bp,编码762个氨基酸,蛋白质序列中含有氧化鲨烯环化酶家族所特有的且高度保守的DCTAE、QW和MWCYCR 3个序列;BsAS无跨膜区,为亲水性蛋白,定位于细胞质中;其二级结构主要为α-螺旋和无规卷曲,三级结构预测结果可信。狭叶柴胡BsAS蛋白共有53个磷酸化位点,其中丝氨酸的磷酸化位点最多。【结论】本研究首次在狭叶柴胡中克隆了BsAS全长基因,同时利用生物信息学分析了其蛋白特性及细胞定位,该研究丰富了BsAS基因资源库,同时为齐墩果烷型代谢产物及代谢途径的深入研究奠定基础。【Objective】The full-length sequence ofβ-amyrin synthase gene and its biological information were obtained in this paper.【Method】The cDNA sequence ofβ-amyrin synthase gene,the key enzyme of oleanolic acid synthesis,which was an effective component of anti-hepatitis,was obtained using Bupleurum scorzonerifolium as tested materials,and the gene was conserved using biological information database and analysis software.The conserved sequence,physicochemical properties,transmembrane domain,subcellular localization,secondary structure,tertiary structure,phosphorylation site and phylogeny were analyzed by using bioinformatics database and analysis software.【Result】The ORF nucleotide length ofβ-amyrin synthase gene was 2289 bp,encoding 762 amino acids.The protein sequence contained three highly conserved DCTAE,QW and MWCYCR sequences unique to the oxidosqualene cyclase family;BsAS had no transmembrane region,which was a hydrophilic protein and located in the cytoplasm;Its secondary structure was mainlyα-helix and random coil,and the prediction of tertiary structure was credible.There were 53 phosphorylation sites in BsAS of Bupleurum scorzonerifolium,of which serine hadthe most phosphorylation sites.【Conclusion】The successful acquisition of the full-length BsAS sequence found a basis for the study of the structure and function of BsAS gene in Bupleurum scorzonerifolium and provided new insights into the regulation of the triterpene synthesis pathway in Bupleurum scorzonerifolium.

关 键 词：狭叶柴胡 β-amyrin合成酶 基因克隆 生物信息学 

分 类 号：S567.23[农业科学—中草药栽培]