An N-terminally fused Xenopus transcription factor IIIA synthesized in Escherichia coli is biologically active.
A 1.5-kilobase DNA fragment containing the Xenopus transcription factor IIIA (TFIIIA) gene was inserted into the prokaryotic expression vector pIN-III(A) containing the lpp/lac promoter. The recombinant DNA was introduced into Escherichia coli K-12 strain SB221. The expression TFIIIA gene was induced by isopropyl-beta-D-thiogalactopyranoside, which resulted in the synthesis of a recombinant TFIIIA with an extra 17 amino acids fused to its N terminus as predicted from the nucleotide sequence. The engineered gene product, purified to at least 90% homogeneity, retained its binding affinity to the intragenic control region of the 5 S RNA gene, as well as its activity to stimulate 5 S RNA gene transcription in vitro.
Pao
C I
CI
Department of Pharmacological Sciences, State University of New York, Stony Brook 11794.
Lee
T C
TC
Liao
Y D
YD
Wu
C W
CW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012335:RNA, Ribosomal / Q000235:genetics*; D012341:RNA, Ribosomal, 5S / Q000235:genetics*; D011994:Recombinant Proteins / Q000302:isolation & purification; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D014981:Xenopus
D012335:RNA, Ribosomal; D012341:RNA, Ribosomal, 5S; D011994:Recombinant Proteins; D035582:Transcription Factor TFIIIA; D014157:Transcription Factors