Armengaud J, de Nuova Perez L, Lemay P, Masson J M
Institut National des Sciences Appliquées, UA 544 du CNRS, Toulouse, France.
FEBS Lett. 1991 Apr 22;282(1):157-60. doi: 10.1016/0014-5793(91)80467-h.
A full length tat gene was constructed by a combination of polymerase chain reaction (PCR) for the first exon and chemical synthesis for the second exon. This gene was expressed in E. coli under the control of the strongly regulated araB promoter, either directly or fused to a secretion signal encoding sequence. We then defined a rapid, three-step procedure for the purification of the Tat protein.
通过聚合酶链反应(PCR)扩增第一个外显子并化学合成第二个外显子,构建了全长tat基因。该基因在强调控的araB启动子控制下于大肠杆菌中表达,既可以直接表达,也可以与编码分泌信号的序列融合表达。然后,我们确定了一种快速的三步法来纯化Tat蛋白。