Genetic transformation in Streptococcus pneumoniae: nucleotide sequence and predicted amino acid sequence of recP

Abstract

We present the complete nucleotide sequence of recP, a locus required for high-efficiency recombination of chromosomal DNA during genetic transformation in Streptococcus pneumoniae. The sequence was determined by using plasmid DNA templates and synthetic oligonucleotide primers. The locus contained a single large open reading frame, ORF1, of 1,968 base pairs (bp). ORF1 is included within the recP locus previously mapped genetically and accounts for 94% of its extent. The predicted molecular weight of the largest polypeptide encoded within ORF1, 71,662, coincided with that measured previously (72,000) for the product of in vitro transcription-translation of the cloned recP locus. A Shine-Dalgarno sequence (AGAAAGGA; delta G = -17 kcal [ca. -71.1 kJ]) lay 6 bp upstream of ORF1. A sequence (TTGcat-17 bp-TATAAT) similar to the E. coli sigma-70 promoter consensus was located 52 bp upstream of ORF1. This putative promoter overlapped a structure consisting of two perfect inverted 10-bp repeats and a loop of 8 bp.