PZR基因的定点突变、表达和纯化  

Site-directed Mutagenesis,Expression and Purification of PZR

作　　者：莫毅[1,2] 王伟[3] 梁方方 杨旭芬[3] 宋茜[3] 蒋和生[1] Wayne Zhou 

机构地区：[1]广西大学动物科学技术学院,南宁530004 [2]广西人口和计划生育研究中心,南宁530021 [3]美国路易斯安那州立大学生命科学院 [4]广西畜牧研究所,南宁530001

出　　处：《中国生物工程杂志》2009年第3期36-40,共5页China Biotechnology

摘　　要：为获得纯化的单磷酸化PZR蛋白,使用双磷酸化位点的His-PZR重组原核表达载体为模板,采用重叠延长PCR定点突变技术分别构建:PZR-Phe263(TTT)和PZR-Phe241(TTT)重组原核表达载体,并与C-Src重组原核表达载体共同转化E.coliBL21菌株,通过IPTG诱导表达,Ni-NTA纯化,获得了单磷酸化的PZRa和PZRb蛋白,为进一步研究PZR内的不同ITIM基序与酪氨酸磷酸酶间的作用机制及其在信号转导过程中的功能奠定了基础。In order to express and purify the mono-phosphorylated PZR proteins, the bi-phosphorylated-site His-PZR recombined prokaryotic expressions plasmids was used as the template, then through the overlapping PCR the mono- phosphorylated-site PZR-Phe263 （TTT） and PZR-Phe241（TTT） mutants were amplified. After recombined and purified, the mutants＇ and C-Src recombined prokaryotic expressions plasmids were cotransformed into the E. Coli BL21. Through IPTG induction and Ni-NTA purification, the mono-phosphorylated PZR a and PZR b proteins were obtained successfully. The results laid a foundation for future study on the PZR bio-function mechanism.

关 键 词：PZR 免疫受体酪氨酸抑制基序 定点突变 表达 纯化 

分 类 号：R392.11[医药卫生—免疫学]