小麦条锈菌效应蛋白基因pst-2511的克隆、 表达和初步结构研究

对小麦条锈菌（Puccinia striiformis f.sp. tritici）效应蛋白pst-2511进行研究，分析其可能形成二级结构的条件，为以后通过核磁方法解析该效应蛋白的三维结构提供基础。构建了小麦条锈菌效应蛋白基因pst-2511的原核表达载体pET-32a-PP-pst-2511，并成功将其在大肠杆菌 BL21(DE3)中进行原核表达；表达出的蛋白用亲和层析法和高效液相色谱法进行纯化，获得可溶性的效应蛋白，用凝胶电泳检测得到单一条带，经质谱鉴定，纯化后的蛋白质分子量与理论值符合。圆二色谱表征该蛋白质的二级结构有α螺旋，且随着三氟乙醇（TFE）浓度的增加，α螺旋比例增加。对效应蛋白pst-2511的初步结构研究及三维结构解析，可以为防止小麦条锈病提供理论依据，同时为深入研究效应蛋白的调控通路以及阐明条锈菌侵染机制和更有效地防止小麦条锈病提供帮助。

Study on cloning, expression and preliminary structure of wheat stripe rust effect protein pst-2511

The effector protein pst-2511 of Puccinia striiformis f.sp. tritici was studied to analyze the possible secondary structure of the target protein for providing a basis for the subsequent analysis on the three-dimensional structure of the effector protein by nuclear magnetic methods. In this study, the prokaryotic expression vector pET-32a-PP-pst-2511 of wheat stripe rust effector gene pst-2511 was constructed, and its prokaryotic expression was successfully performed in E. coli BL21 (DE3); the protein expressed was purified by affinity chromatography and high-performance liquid chromatography to obtain soluble effector protein. A single band was detected by gel electrophoresis. The mass of the purified protein was consistent with the theoretical value by mass spectrometry. The secondary structure of the protein was characterized by a circular dichroism, and the alpha helix was increased with the increase of the concentration of trifluoroethanol (TFE). The preliminary structure study and three-dimensional structure analysis of the effector protein pst-2511 can provide a theoretical basis for the prevention of wheat stripe rust. Meanwhile, it provides an in-depth study on the regulatory pathways of effector proteins and elucidates the infection mechanism of stripe rust and more effective prevention of wheat strips rust.