Northern Illinois University, Biological Sciences, Dekalb, IL 60115
Transcriptome analysis of Neisseria gonorrhoeae, the causative agent of the human sexually transmitted disease gonorrheae, has revealed accumulations of mRNA transcripts along certain areas of the pilE gene, especially in the 5’ and mid-gene segments. Northern blotting of E.coli transformants containing a cloned segment of the mid-gene region, as well as in vitro transcription of the mid-gene segment, have indicated the presence of sense and antisense transcripts originating in the mid-gene region of pilE. Furthermore, strand-specific qRT-PCR of full-segment pilE clones in E.coli have yielded results indicating an accumulation of antisense RNA transcript at the 3’ end of the gene, as well as confirming other previously found putative promoters. Current research has been focused on confirming the location of the mid-gene putative promoter using site-directed mutagenesis followed by Northern blotting and qRT-PCR. Also, strand-specific qRT-PCR assays of total RNA extracted directly from a strain of Neisseria gonorrhoeae are underway to confirm the utilization and transcription of the putative promoters in vivo.
[Abstract (DOCX)]