马立克氏病毒UL36蛋白的表达及其去泛素化酶活性分析

发布时间：2018-03-03 19:54

  本文选题：MDV　切入点：UL　出处：《畜牧与兽医》2017年05期 　论文类型：期刊论文

【摘要】：选取马立克氏病毒(MDV)去泛素化酶UL36蛋白N端具有蛋白酶活性的一段氨基酸序列,表达纯化并分析UL36的去泛素化酶活性特点。利用p Cold TF为表达载体,并在UL36-323的C端连接Strep tagⅡ作为纯化标签,构建重组质粒p Cold TF-UL36-Strep。应用大肠杆菌表达体系,低温诱导表达,并对该段蛋白进行纯化,得到可溶且纯度较高的蛋白后,通过Western blot确定所纯化的蛋白即是UL36蛋白。应用Di-Ub(K48)和SUMO1-GST作为底物鉴定其去泛素化酶活性。最终得到了可溶且纯度较高的UL36-323-Strep蛋白,并且通过酶切反应确定了其去泛素化酶活性,为进一步探究UL36在MDV感染及复制中的作用奠定前期基础,也为探究MDV的致病机理提供了必要的前提条件。
[Abstract]:A sequence of amino acids with protease activity at the N-terminal of UL36 protein of Marek's virus (MDV) was selected to express, purify and analyze the characteristics of UL36 activity. The expression vector was p Cold TF. The recombinant plasmid p Cold TF-UL36-Strep. expressed in Escherichia coli was induced to express at low temperature, and the protein was purified to obtain soluble and high purity protein, and the recombinant plasmid p#en3# TF-UL36-Strep. was constructed by ligating Strep tag 鈪，

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