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拟南芥O-岩藻糖基转移酶AtPOFUT1的鉴定及生理功能初探
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引用本文:梁 蓉,谢尚强,赵小明,贾晓晨,尹 恒.拟南芥O-岩藻糖基转移酶AtPOFUT1的鉴定及生理功能初探[J].西北农业学报,2021,(1):116~123
DOI:10.7606/j.issn.1004-1389.2021.01.014
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作者单位
梁 蓉,谢尚强,赵小明,贾晓晨,尹 恒 (1.大连市糖类农用制剂工程研究中心,辽宁省碳水化合物研究重点实验室,中国科学院大连化学物理研究所辽宁大连 116023
2.中国科学院大学,北京 100049) 
基金项目:国家自然科学基金(31672077);大连化物所创新基金(DICPI201905)。
中文摘要:O-岩藻糖基转移酶(protein O-fucosyltransferase,POFUT)是催化蛋白质O-岩藻糖基化的关键酶,被证实与动物的生理病理过程密切相关。但在植物中,POFUT的鉴定和功能研究尚处于起步阶段。首先通过同源序列的BLAST和生物信息学分析,推测在拟南芥基因组中AT3G05320编码的蛋白具有潜在的POFUT活性,并将其命名为AtPOFUT1。利用毕赤酵母真核表达体系实现AtPOFUT1蛋白的异源表达,初步检测明确该重组蛋白具有POFUT活性。进一步利用公共基因芯片数据对AtPOFUT1表达模式进行分析,发现AtPOFUT1在植物的生长发育和抗病抗逆等过程发挥重要作用。对AtPOFUT1缺失突变体的表型统计分析,发现突变体表现出种子数量减少且发育不良等性状,明确AtPOFUT1在拟南芥生长发育中的重要功能。上述研究初步揭示AtPOFUT1的催化活性及部分生理功能,可为植物中POFUT的鉴定及生物学功能研究工作提供一定参考。
中文关键词:O-岩藻糖基化  POFUT  拟南芥  异源表达  生理功能
 
Identification and Physiological Function Investigation of Arabidopsis O-fucosyltransferase AtPOFUT1
Abstract:O-fucosyltransferase (POFUT) is a key enzyme that catalyzes protein O-fucosylation, it isproved that it had close relation with the physiological and pathological processes of animals, however, the identification and function investigation of POFUT in plants are still in its infancy. In this study, by using BLAST and bioinformatics analysis, AT3G05320 in the Arabidopsis genome was selected and was named AtPOFUT1.Heterologous expression of AtPOFUT1 protein was successfully achieved by using the Pichia pastoris expression system, and preliminary detection had confirmed that the recombinant protein had POFUT activity. Further analysis of AtPOFUT1 expression patterns using public gene chip data revealed that AtPOFUT1 played an important role in plant growth and development, disease resistance and stress tolerance. The statistical analysis of the phenotypes of Atpofut1 mutant plants revealed that the mutants exhibited traits such as reduction of seed number and dysplasia, and the important functions of AtPOFUT1 in growth and development was clarified. The studies above revealed the catalytic activity and some physiological functions of AtPOFUT1, which could provide references for identification of POFUT in plnats and research of its biological functions.
keywords:O-fucosylation  POFUT  Arabidopsis thaliana  Heterologous expression  Physiological function
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