Organization of the Thermus thermophilus nusA/infB operon and overexpression of the infB gene in Escherichia coli.
The structural gene for translation initiation factor IF2 from Thermus thermophilus was identified on the basis of the N-terminal amino acid sequence of intact T thermophilus IF2 and an internal 25 kDa IF2 fragment. A total of 5135 bp was cloned and sequenced, comprising the open reading frames for p15A, NusA, p10A, IF2, p10B and SecD, which may form an operon. There are pronounced similarities between the operon arrangement and primary sequence of the T thermophilus genes and proteins, respectively, and their counterparts from other organisms. The T thermophilus infB gene was expressed to a high level in E coli. Four hundred milligrams of homogenous T thermophilus IF2 were prepared from 60 g of overproducing cells.
Vornlocher
H P
HP
Laboratorium für Biochemie, Universität Bayreuth, Germany.
Kreutzer
R
R
Sprinzl
M
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D015870:Gene Expression; D008969:Molecular Sequence Data; D009876:Operon; D010445:Peptide Elongation Factors; D010448:Peptide Initiation Factors / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D039881:Prokaryotic Initiation Factor-2; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D016963:Thermus thermophilus / Q000235:genetics*; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D035602:Transcriptional Elongation Factors
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D010445:Peptide Elongation Factors; D010448:Peptide Initiation Factors; D039881:Prokaryotic Initiation Factor-2; D011994:Recombinant Proteins; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; C068727:nusA protein, E coli