Molecular cloning and sequence of the Bacillus stearothermophilus translational initiation factor IF2 gene

Molecular & General Genetics : MGG
M BrombachC L Pon

Abstract

The structural gene for the Bacillus stearothermophilus initiation factor IF2 was localized to a 6 kb HindIII restriction fragment by cross-hybridization with the SstI-SmaI fragment of the Escherichia coli infB gene. This fragment corresponds to the central region of the molecule containing the GTP-binding domain which is homologous in E. coli IF2, EF-Tu, EF-G and the human ras1 oncogene protein. After cloning into pACYC177, the HindIII fragment was further analysed by restriction mapping and cross-hybridization. A smaller (2.2 kb) SphI-HindIII fragment, which showed cross-hybridization, was subcloned into M13 phage and sequenced by the dideoxy chain-terminating method. This fragment was found to contain the entire IF2 gene except for the region coding for the N-terminus. This remaining region, coding for 45 amino acids, was located by homologous hybridization on an overlapping ClaI-SstI fragment which was also subcloned and sequenced. Overall, the B. stearothermophilus IF2 gene codes for a protein of 742 amino acids (Mr = 82,043) whose primary sequence displays extensive homology with the C-terminal two-thirds (but little or no homology with the N-terminal one-third) of the corresponding E. coli IF2 molecule. When cloned into ...Continue Reading

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Jan 15, 1997·European Journal of Biochemistry·H P VornlocherMathias Sprinzl
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Related Concepts

Geobacillus stearothermophilus
DNA Sequence
DNA Restriction Enzymes
Alkalescens-Dispar Group
Cistron
Genes, Bacterial
Genomic Hybridization
Peptide Initiation Factors
Gene Products, Protein
Eukaryotic Initiation Factor-2, gamma Subunit

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