Production of a full length Tat protein in E. coli and its purification.
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.
Armengaud
J
J
Institut National des Sciences Appliquées, UA 544 du CNRS, Toulouse, France.
de Nuova Perez
L
L
Lemay
P
P
Masson
J M
JM
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral; D004926:Escherichia coli / Q000235:genetics*; D005091:Exons; D015696:Gene Products, tat / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005813:Genes, Synthetic; D005814:Genes, Viral; D015756:Genes, tat; D015497:HIV-1 / Q000235:genetics*; D007700:Kinetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D054322:tat Gene Products, Human Immunodeficiency Virus
D004279:DNA, Viral; D015696:Gene Products, tat; D054322:tat Gene Products, Human Immunodeficiency Virus