本文已被：浏览 1408次 下载 513次
投稿时间：2019-03-21
投稿时间：2019-03-21
中文摘要: 多酚氧化酶是普遍存在于植物、动物、细菌及真菌中的一类由核基因编码的具有氧化酚类底物成醌类作用的金属铜蛋白酶，也是褐变过程中的关键酶类。本研究通过对马铃薯基因组数据库检索，发现马铃薯多酚氧化酶包含StuPPO1 到StuPPO9 中9 个基因成员，其中StuPPO5-9 为预测基因。同时StuPPO9 是生物和非生物因素诱导表达的基因，然而其序列和功能未被验证。本试验以“大西洋”马铃薯叶片为试材，分别提取总RNA 和gDNA，对该基因进行分离鉴定，同时对其进行过表达载体构建和转基因烟草转化，为后续基因功能验证提供技术支持。试验结果表明，StuPPO9 基因大小为1 884 bp，与XM_006347021 一致性为98.4%，该基因含有一个内含子结构，与其他成员不同，它位于2 号染色体上。该基因启动子可能被生物和非生物因素诱导激活。利用农杆菌介导的基因过表达技术，将该基因转入烟草中，通过聚合酶链式反应（polymerase chain reaction，PCR）检测获得了阳性株系。
Abstract:Polyphenol oxidase is a kind of metal copper protease which is ubiquitously present in plants，animals，bacteria and fungi and which is encoded by a nuclear gene and has a oxidized phenolic substrate to form a quinone.It is also a key enzyme in the browning process.In this study，it searched the potato genome database and found that potato polyphenol oxidase contains nine gene members from StuPPO1 to StuPPO9，which StuPPO5-9 was a predictor gene.At the same time，StuPPO9 was a gene induced by biotic and abiotic factors，however its sequence and function had not been verified.In this experiment，the “Atlantic”potato leaves were used as test materials to extract total RNA and gDNA and the gene was isolated and identified.At the same time，the expression vector construction and transgenic tobacco transformation were carried out to provide technical support for subsequent gene function verification.The results showed that the size of StuPPO9 gene was 1 884 bp，and the identity with XM_006347021 was 98.4%.The gene contained an intron structure ，unlike other members was located on chromosome 2.The gene promoter might be activated by biotic and abiotic factors.The gene was transferred into tobacco using agrobacterium-mediated gene overexpression technology and obtained a positive strain by polymerase chain reaction（PCR）.
文章编号：202002029 中图分类号： 文献标志码：
基金项目:国家自然基金青年计划项目（31902094、31801682）；天津市教委科研计划项目（2018KJ192）；天津农学院科学研究发展基金计划项目（2016NZD01）；天津市国家级大学生创新创业训练计划项目（201710061037）
引用文本：
津ICP备20000603号-1