Rapid detection of biothreat agents based on cellular machinery.

Lane, Todd W. ; Gantt, Richard W.

Sandia National Laboratories

关键词: Shigella;    Biological Detection.;    Testing;    Rna Polymerases;    Genes;   

DOI  :  10.2172/920835
RP-ID  :  SAND2004-6381
RP-ID  :  AC04-94AL85000
RP-ID  :  920835

美国|英语

来源: UNT Digital Library

【 摘 要 】

This research addresses rapid and sensitive identification of biological agents in a complex background. We attempted to devise a method by which the specificity of the cellular transcriptional machinery could be used to detect and identify bacterial bio-terror agents in a background of other organisms. Bacterial cells contain RNA polymerases and transcription factors that transcribe genes into mRNA for translation into proteins. RNA polymerases in conjunction with transcription factors recognize regulatory elements (promoters) upstream of the gene. These promoters are, in many cases, recognized by the polymerase and transcription factor combinations of one species only. We have engineered a plasmid, for Escherichia coli, containing the virA promoter from the target species Shigella flexneri. This promoter was fused to a reporter gene Green Fluorescent Protein (GFP). In theory the indicator strain (carrying the plasmid) is mixed with the target strain and the two are lysed. The cellular machinery from both cells mixes and the GFP is produced. This report details the results of testing this system.