WANG Huizheng,LAN Yubin.Expression，Purification and Crystallization of Carnitine Acyl Transferase From Phytophthora capsica[J].Northern Horticulture,2020,44(11):42-46.[doi:10.11937/bfyy.20193355]
辣椒疫霉菌肉碱脂酰转移酶基因CAT的表达、蛋白纯化及结晶
- Title:
- Expression，Purification and Crystallization of Carnitine Acyl Transferase From Phytophthora capsica
- Keywords:
- Phytophthora capsici; carnitine acyl transferase; expression; purification; crystallization
- 文献标志码:
- A
- 摘要:
- 肉碱脂酰转移酶CAT是与附着胞的形成紧密相关的一类酶，与辣椒疫霉菌的致病性密切相关。该研究基于辣椒疫霉菌全基因组序列，以辣椒疫霉高致病菌株SD33全基因组DNA为试材扩增CAT基因，采用了原核表达方法诱导表达重组蛋白6×His-CAT，并对重组蛋白进行亲和层析纯化、分子排阻层析纯化及蛋白结晶试验，以期获得纯蛋白及蛋白晶体。结果表明：经上述方法所获蛋白具有较高的纯度，且有较好的结晶性能，为进一步阐明CAT作用机理奠定基础。
- Abstract:
- Carnitine acyl transferase CAT is a kind of enzymes closely related to appressorium formation and pathogenicity of Phytophthora capsici.Based on the whole-genome sequence data of P.capsici，the whole genome DNA of highly pathogenic strain P.capsici SD33 was extracted and used to amplify CAT gene.The recombinant protein 6×His-CAT was induced and expressed by prokaryotic expression method，and the recombinant protein was purified by affinity chromatography，molecular exclusion chromatography.The crystallization experiment using purified protein was carried out in order to obtain protein crystals.The results showed that the protein obtained by the above method had high purity and good crystallinity，provided a basis to clarify the mechanism of CAT.
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备注/Memo
第一作者简介：王会征（1984-），男，博士，讲师，现主要从事植物病害及其致病机理等研究工作。E-mail：hzwang@sdut.edu.cn.基金项目：山东省引进顶尖人才“一事一议”专项经费资助项目；淄博市科技发展计划资助项目（2018kj010073）。收稿日期：2019-12-06