The nucleotide sequences defining the signal peptides of the galactose-binding protein and the arabinose-binding protein.

J B Scripture,R W Hogg

doi:10.1016/s0021-9258(17)44353-5

Abstract

The nucleotide sequences of the NH2-terminal regions of the mglB (galactose-binding protein) and the araF (arabinose-binding protein) genes have been determined. A "signal peptide" containing 23 amino acid residues immediately precedes the known sequence of the processed galactose-binding protein. The primary sequence of the signal peptide of the arabinose-binding protein has been redefined according to the established nucleotide sequence. The nucleotide sequences for the two signal peptides are preceded by apparent ribosome-binding sequences.

Highlights

The nucleotide sequences of theNH2-terminal re- galactose-binding protein and its derived amino acid sequence gions of the mglB and the and thebase sequence of the arabinose-binding proteinsignal araFgenes have been de- region
The primary Growth Conditwns and Organisms-Bacterial cells were cultured sequence of the signal peptide of the arabinose-binding in L broth (Luriaand Burrows, 1957)or yeast tryptone brothat 37 “C protein has been redefined according to the establishoenda roller rack or in a shaking water bath
Source of DNA-DNA fragments for sequencing the galactosebinding proteinsignal region wereobtained from the plasmid pMG10, a derivative of the plasmid pMG3, described by Rotman and Guzman (1982)

Summary

Bethesda Research

P-L Biochemicals P-L Biochemicals method of Hogg (1971). A 6.3-kilobase plasmid, pAT1, was isolated and a restriction map generated (Figs. 1 and 2). A 6.3-kilobase plasmid, pAT1, was isolated and a restriction map generated Plasmid PAT1 was digested with SmaI and HpaI resulting in two fragments, 6000 and 260 base pairs, respectively. The fragments were ligated into SrnaIdigested M13mp and used to transformE. ATlshl and ATlsh, containing the 260-base pair fragment were selected for sequencing. Plasmid PAT1was digested with SmaI andEcoRI and thesmaller 1150-base pair fragment was isolated anddesignatedATlse.The purified DNA fragment, ATlse,was digested with Sau3A (compatible with BarnHI-generatedends), ligated intoBamHI, EcoRI-digested M13mp, andthemixture used totransform E. coli JM103.One clone, ATlxe was selected for sequencing. The Sma1:EcoRI fragment,ATlse, wasdigestedwithAluI and ligated in the presence of SmaI-digested M13mp. Following transformation of E. coli JM103, two complementing clones, Atlaal and ATlaa, were selected for sequencing.Asecond sample of ATlse, alsodigested with AluI,wasligated inthe presence of M13mp previously digested with SrnaI and EcoRI. E. coli JM103 was transformedwith themixtureand aclone, ATlaelO, was selected for sequencing

RESULTS AND DISCUSSION

TTA Val L e u

Val L y s Gln P r oG l u Glu