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J Bacteriol. 1991 December; 173(24): 7896-7902

research-article

Analysis of the lsi region involved in lipooligosaccharide biosynthesis in Neisseria gonorrhoeae.

E F Petricoin 3rd, R J Danaher and D C Stein

Department of Microbiology, University of Maryland, College Park 20742.

ABSTRACT

The genetic locus (lsi-1) responsible for the transformation of the lipooligosaccharide (LOS)-defective Neisseria gonorrhoeae mutant FA5100 to LOS expression was studied by deletion mutagenesis and sequence analysis. An open reading frame that was preceded by a leader sequence containing regions with the potential to form hairpin loops was identified. A perfect sigma 70 promoter consensus sequence was found upstream from this open reading frame. Promoter function was screened for functionality by using lac fusion cassettes and in vitro transcription-translation analysis. A frameshift mutation in the lsi-1 gene was constructed by site-directed mutagenesis and introduced into the chromosome of FA19, the LOS-expressing isogenic parent strain of FA5100. The mutant was characterized by Southern blotting, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Western blotting (immunoblotting) and found to be phenotypically identical to FA5100.

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J Bacteriol. 1991 December; 173(24): 7896-7902

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