USE OF BACTERIAL LUCIFERASE STRUCTURAL GENES FOR CLONING AND MONITORING GENE EXPRESSION IN MICROORGANISMS AND FOR TAGGING AND IDENTIFICATION OF GENETICALLY ENGINEERED ORGANISMS

摘要

A host microorganism is genetically and stably modified by the insertion in any of its non-essential chromosomal locations a foreign DNA fragment comprising a non-homologous fragment containing a lux AB gene of a selected bioluminescent bacterium such as V. harveyi such that the nonhomologous fragment is stably reproduced as a part of the chromosome of the cells of the host microorganism as they divide in a growth medium and the gene expression of such microorganism triggers the production of luciferase which, in turn emits light in the presence of aldehyde as a measure of gene expression. Both Gram-positive and Gram-negative bacteria are microorganisms which may be modified as aforesaid. X-ray film can be used to quantify the light being emitted from a microorganism through the use of plural droplets containing the same microorganism each with a known and related cell (or plasmid) count.