@article {Hillman747980, author = {Tatiana Hillman}, title = {Integrating the metabolic processes of Escherichia coli with virulence by decreasing glucose availability, inhibiting the acetyl-CoA carboxylase gene accA with asRNA, and through the quantification of the luxs gene}, elocation-id = {747980}, year = {2019}, doi = {10.1101/747980}, publisher = {Cold Spring Harbor Laboratory}, abstract = {The study aims to demonstrate a possible link between bacterial cell metabolism and virulence by combining the metabolic mechanisms of the gram-negative bacteria, Escherichia coli. Glucose increases the proliferation of intestinal microflora, which augments the output of the short-chain fatty acids. Bacteria ferment glucose into short-chain fatty acids, which help regulate many biochemical processes and pathways. Each short-chain fatty acid maintains colonic pH, promotes cell differentiation, and the apoptosis of colonocytes. The long-chain fatty acids are also synthesized for plasma membrane and biofilm formation. To increase the synthesis of acetyl-CoA carboxylase, an enzyme that catabolizes glucose into short- and long-chain fatty acids, Escherichia coli was cultured in Luria broth enhanced with a high to a low concentration of glucose. The 15mM, a high concentration of glucose, yielded qPCR products measured for the target gene accA, which was 4,210ng/{\textmu}L. The 7.5mM sample produced a concentration equal to 375 ng/{\textmu}L, and the control sample measured an accA concentration of 196 ng/{\textmu}L. The gene accA, one of four subunits for the acetyl-CoA carboxylase enzyme, was suppressed by asRNA, producing a qPCR product of 63 gene copies. Antisense RNA for accA reduced the amount of luxs, a vital gene needed for propagating quorum-sensing signal molecules. The luxs gene, which is responsible for releasing autoinducer 2 for cell-to-cell quorum sensing, was reduced by the gene inhibition of accA with asRNA. The increase in luxs transcription augments biofilm production in support of spreading virulence. The implications of the study advocate for designing antibiotics that target bacterial cell metabolic processes to block bacterial antibiotic resistance.}, URL = {https://www.biorxiv.org/content/early/2019/09/18/747980}, eprint = {https://www.biorxiv.org/content/early/2019/09/18/747980.full.pdf}, journal = {bioRxiv} }