Abstract
Previously, we observed butanal/propanal sensitivity of Escherichia coli K-12 when cells overexpress YqhD protein, a NADPH dependent aldehyde reductase, possibly due to an accumulation of butanol/propanol in vivo as the reaction products. Based on this finding, we developed a suicidal vector-cloning system derived from pUC19, in which lacZ was substituted with the yqhD gene. As a result, when foreign DNA was inserted into its multiple cloning sites by disrupting an expression of YqhD, the recombinants survived on butanal/propanal containing plate, whereas cells containing the YqhD vector died because of the alcohol production by YqhD. The cloning efficiency, estimated based on colony PCR and enzyme digestion, was achieved more than 90% when the suicidal vector system was used. Moreover, the plasmid vector itself was stably maintained in the cell, presumably due to its ability to remove toxic aldehydes being accumulated in E. coli cell by metabolic stress.
Similar content being viewed by others
References
Anne, I., Goma, G., and Soucaille, P. 1989. Effects of various alcoholic supplements on the growth rate of Clostridium acetobutylicum ATCC 824. Appl. Microbiol. Biotechnol. 31, 179–183.
Bernard, P. and Couturier, M. 1992. Cell killing by the F plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes. J. Mol. Biol. 226, 735–745.
Bernard, P., Gabant, P., Bahassi, E.M., and Couturier, M. 1994. Positive-selection vectors using the F plasmid ccdB killer gene. Gene 148, 71–74.
Bernard, P., Kezdy, K.E., Van Melderen, L., Steyaert, J., Wyns, L., Pato, M.L., Higgins, P.N., and Couturier, M. 1993. The F plasmid CcdB protein induces efficient ATP-dependent DNA cleavage by gyrase. J. Mol. Biol. 234, 534–541.
Burioni, R., Plaisant, P., Riccio, M.L., Rossolini, G.M., and Satta, G. 1995. A new plasmid cloning vector for direct detection of recombinant clones, based on inactivation of a bacterial acid phosphatase-encoding gene. New Microbiol. 18, 201–206.
Chaffin, D.O. and Rubens, C.E. 1998. Blue/white screening of recombinant plasmids in Gram-positive bacteria by interruption of alkaline phosphatase gene (phoZ) expression. Gene 219, 91–99.
Clark, E. and Cirvilleri, G. 1994. Cloning cassettes containing the reporter gene xylE. Gene 151, 329–330.
Dedonder, R. 1966. Levansucrase from Bacillus subtilis. Methods Emzymol. 8, 500–505.
Friehs, K. 2004. Plasmid copy number and plasmid stability. Adv. Biochem. Eng. Biotechnol. 86, 47–82.
Gabant, P., Szpirer, C.Y., Couturier, M., and Faelen, M. 1998. Direct selection cloning vectors adapted to the genetic analysis of Gram-negative bacteria and their plasmids. Gene 207, 87–92.
Gay, P., Le Coq, D., Steinmetz, M., Berkelman, T., and Kado, C.I. 1985. Positive selection procedure for entrapment of insertion sequence elements in Gram-negative bacteria. J. Bacteriol. 164, 918–921.
Gay, P., Le Coq, D., Steinmetz, M., Ferrari, E., and Hoch, J.A. 1983. Cloning structural gene sacB, which codes for exoenzyme levansucrase of Bacillus subtilis: expression of the gene in Escherichia coli. J. Bacteriol. 153, 1424–1431.
Goh, S. and Good, L. 2008. Plasmid selection in Escherichia coli using an endogenous essential gene marker. BMC Biotechnol. 8, 61.
Henrich, B. and Schmidtberger, B. 1995. Positive-selection vector with enhanced lytic potential based on a variant of phi X174 phage gene E. Gene 154, 51–54.
Inouye, S., Ogawa, H., Yasuda, K., Umesono, K., and Tsuji, F.I. 1997. A bacterial cloning vector using a mutated Aequorea green fluorescent protein as an indicator. Gene 189, 159–162.
Jager, W., Schafer, A., Puhler, A., Labes, G., and Wohlleben, W. 1992. Expression of the Bacillus subtilis sacB gene leads to sucrose sensitivity in the Gram-positive bacterium Corynebacterium glutamicumbut not in Streptomyces lividans. J. Bacteriol. 174, 5462–5465.
Jarboe, L.R. 2011. YqhD: a broad-substrate range aldehyde reductase with various applications in production of biorenewable fuels and chemicals. Appl. Microbiol. Biotechnol. 89, 249–257.
Jean, B., Masion, E., Amine, J., Petitdemange, H., and Gay, R. 1987. Inhibitor effect of products of metabolism on growth of Clostridium acetobutylicum. Appl. Microbiol. Biotechnol. 26, 568–573.
Kasper, M., Roehlecke, C., Witt, M., Fehrenbach, H., Hofer, A., Miyata, T., Weigert, C., Funk, R.H., and Schleicher, E.D. 2000. Induction of apoptosis by glyoxal in human embryonic lung epithelial cell line L132. Am. J. Respir. Cell Mol. Biol. 23, 485–491.
Lee, C., Kim, I., Lee, J., Lee, K.L., Min, B., and Park, C. 2010. Transcriptional activation of the aldehyde reductase YqhD by YqhC and its implication in glyoxal metabolism of Escherichia coli K-12. J. Bacteriol. 192, 4205–4214.
Miller, E.N., Jarboe, L.R., Yomano, L.P., York, S.W., Shanmugam, K.T., and Ingram, L.O. 2009. Silencing of NADPH-dependent oxidoreductase genes (yqhD and dkgA) in furfural-resistant ethanologenic Escherichia coli. Appl. Environ. Microbiol. 75, 4315–4323.
Mlakar, A., Batna, A., Dudda, A., and Spiteller, G. 1996. Iron (II) ions induced oxidation of ascorbic acid and glucose. Free Radic. Res. 25, 525–539.
Pelicic, V., Reyrat, J.M., and Gicquel, B. 1996. Expression of the Bacillus subtilis sacB gene confers sucrose sensitivity on mycobacteria. J. Bacteriol. 178, 1197–1199.
Perez, J.M., Arenas, F.A., Pradenas, G.A., Sandoval, J.M., and Vasquez, C.C. 2008. Escherichia coli YqhD exhibits aldehyde reductase activity and protects from the harmful effect of lipid peroxidation-derived aldehydes. J. Biol. Chem. 283, 7346–7353.
Riccio, M.L., Rossolini, G.M., Lombardi, G., Chiesurin, A., and Satta, G. 1997. Expression cloning of different bacterial phosphatase-encoding genes by histochemical screening of genomic libraries onto an indicator medium containing phenolphthalein diphosphate and methyl green. J. Appl. Microbiol. 82, 177–185.
Sambrook, J. and Russel, D.W. 2001. Molecular cloning; A labo ratory manual, 3rd ed. CSHL press, New York, N.Y., USA.
Schlieper, D., von Wilcken-Bergmann, B., Schmidt, M., Sobek, H., and Muller-Hill, B. 1998. A positive selection vector for cloning of long polymerase chain reaction fragments based on a lethal mutant of the crp gene of Escherichia coli. Anal. Biochem. 257, 203–209.
Steinmetz, M., Le Coq, D., Djemia, H.B., and Gay, P. 1983. Genetic analysis of sacB, the structural gene of a secreted enzyme, levansucrase of Bacillus subtilis Marburg. Mol. Gen. Genet. 191, 138–144.
Sulzenbacher, G., Alvarez, K., Van Den Heuvel, R.H., Versluis, C., Spinelli, S., Campanacci, V., Valencia, C., Cambillau, C., Eklund, H., and Tegoni, M. 2004. Crystal structure of E. coli alcohol dehydrogenase YqhD: evidence of a covalently modified NADP coenzyme. J. Mol. Biol. 342, 489–502.
Thornalley, P.J. 2002. Glycation in diabetic neuropathy: characteristics, consequences, causes, and therapeutic options. Int. Rev. Neurobiol. 50, 37–57.
Van Melderen, L., Bernard, P., and Couturier, M. 1994. Lon-dependent proteolysis of CcdA is the key control for activation of CcdB in plasmid-free segregant bacteria. Mol. Microbiol. 11, 1151–1157.
Wells-Knecht, K.J., Zyzak, D.V., Litchfield, J.E., Thorpe, S.R., and Baynes, J.W. 1995. Mechanism of autoxidative glycosylation: identification of glyoxal and arabinose as intermediates in the autoxidative modification of proteins by glucose. Biochemistry 34, 3702–3709.
Yazynin, S., Lange, H., Mokros, T., Deyev, S., and Lemke, H. 1999. A new phagemid vector for positive selection of recombinants based on a conditionally lethal barnase gene. FEBS Lett. 452, 351–354.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, C., Park, C. Development of a suicidal vector-cloning system based on butanal susceptibility due to an expression of YqhD aldehyde reductase. J Microbiol. 50, 249–255 (2012). https://doi.org/10.1007/s12275-012-1438-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12275-012-1438-5