Bacillus subtilis transformation

Xanthine oxidase, a widely used source of superoxide, has been frequently applied for the study of the effects of superoxide on DNA oxidation. Rozenberg-Arska et al. [30] have shown that xanthine oxidase plus excess iron induced chromosomal and plasmid DNA injury, which was supposedly mediated by hydroxyl radicals. Ito et al. [31] compared the inactivation of Bacillus subtilis transforming DNA by potassium superoxide and the xanthine xanthine oxidase system. It was found that xanthine oxidase but not K02 was a source of free radical mediated DNA inactivation apparently due to the conversion of superoxide to hydroxyl radicals in the presence of iron ions. Deno and Fridovich [32] also supposed that the single strand scission formation after exposure of DNA plasmid to xanthine oxidase was mediated by hydroxyl radical formation. Oxygen radicals produced by xanthine oxidase induced DNA strand breakage in promotable and nonpromotable JB6 mouse epidermal cells [33]. 

Bacillus subtilis transformants were identified by their resistance to naphthalene, parachlorobiphenyl, or dibenzofuran. Controls subtilis protoplasts regenerated without the addition of B. megaterium plasmid DNA) exhibited no increased reistance to these three compounds. That each transformant was B. subtilis and not a contaminant was verified by testing for the B. subtilis chromosomal markers. Only streptomycin resistant strains that showed arginine and leucine dependent growth were used in subsequent experiments. 

Mahler, I. (1968). Procedures for Bacillus subtilis transformation. Methods Enzymol 12B 846. 

Wilson, G. A., and Bott, K. F. (1968). Nutritional factors influencing the development of competence in the Bacillus subtilis transformation system. J Bacteriol 95 1439. 

Early studies by Terawaki and Greenberg on the antibiotic activity of carzinophilin established that it inhibited DNA synthesis but not RNA or protein synthesis in E. coli strain Bo and in Bacillus subtilis [134]. They also found that exposure to carzinophilin removed the transforming capacity of B. subtilis DNA [135]. They... 

Aniline transformed the Balb/3T3 mouse cell line at doses of 0.8 to 100 /dose-response effect), but not the Syrian hamster embryo cells (Dunkel et al. 1981). Results were negative in DNA damage assays in Escherichia coli (Mamber et al. 1983) and Bacillus subtilis (McCarroll et al. 1981). 

Khanna, M. Stotzky, G. (1992). Transformation of Bacillus subtilis by DNA bound on montmorillonite and effect of DNase on the transforming ability of bound DNA. Applied and Environmental Microbiology, 58, 1930-9-... 

There are also data that indicate that aluminum does not directly interact with DNA in mutagenicity tests. These data come from negative transformation assays in Syrian hamster cells (DiPaolo and Casto 1979), negative rec (recombination repair) assays in Bacillus subtilis (Kanematsu et al. 1980), and negative Ames assays in Salmonella typhimurium (Marzin and Phi 1985). These data are summarized in Table 2-4. 

Ohse, M., K. Kawade, and H. Kusaoke. 1997. Effects of DNA topology on transformation efficiency of Bacillus subtilis ISW1214 by electroporation. Biosci. Bio-technol. Biochem. 61 1019-1021. 

The next important property of PolyP-PHB complexes is their effect on DNA transfer into bacterial cells. It was the striking correlation between PolyP-PHB concentrations and transformation efficiencies in Azotobacter vinelandii, Bacillus subtilis and E. coli that led Reusch and Sadoff (1988) to postulate that the complexes are involved in DNA transmembrane transport. 

Artificial transformation of Escherichia coli cells will be performed as part of Experiment 21. In this experiment, you will study natural transformation using strains of the bacterium Bacillus subtilis. Natural transformation has been subjected to extensive study by genetic and biochemical analysis. B. subtilis cells are not able to take up DNA and undergo transformation under all growth conditions, but generally develop competence during periods of nutrient limitation (along with other responses such... 

Anagnostopoulos, C., and Spizizen, J. (1961). Requirements for transformation in Bacillus subtilis. J Bacte-riol 81 741. 

Lotareva, O. V. Prozorov, A. A. (2000). Effect of the clay minerals montmorillonite and kaolinite on the generic transformation of competent Bacillus subtilis cells. Microbiology, 69, 571-4. 

A homologue of the E. coli hisC gene was isolated by transformation of the his, arg" Haloferax volcanii strain WR256 with cosmid DNAs from the Haloferax volcanii minimal set. The deduced amino-acid sequence can be aligned with corresponding bacterial and eucaryal enzymes, with which it is identical at approximately 20 (yeast) or 30 E. coli or Bacillus subtilis) percent of positions. E. coli and B. subtilis are in turn only 30% identical [65]. The strain had previously been shown to be transformable... 

Amin, N.S. Wong, S. Schellenberger, V. Direct transformation of site-saturation libraries in Bacillus subtilis. Biotechniques 2003, 35, 1134-1140. 

In general, sodium selenite and sodium selenate have produced mixed results in bacterial mutagenicity test systems (Table 3M). Sodium selenite induced base-pair substitution mutations using S. typhimurium and was also positive in the transformation assay using Bacillus subtilis (Kramer and Ames 1988 Nakamuro et al. 1976 Noda et al. 1979). However, negative results have also been reported for sodium selenite both in S. typhimurium and the rec assay using B. subtilis (Lofroth and Ames 1978 Noda et al. 

Many of these transformations are also carried out on the analogous anthracyclinones—the aglycone portions of these molecules. Others occur on the glycosides, such as the N-acetylation of the daunosamine moiety of daunomycin and daunomycinol by Bacillus subtilis var, mycoides (36). 

Early attempts to produce XI in S. cerevisiae have failed. Transformation with Actinoplanes missouriensis [53] and Clostridium thermosulfurogenes [55] xylA did not result in the expression of XI, although the specific mRNA was present. The heterologous expression of the E. coli [51,52] and the Bacillus subtilis [53]... 

Microbial transformations also occupy a very important position in the synthesis of enantiomerically pure compounds. Mori and Akao utilized the selective hydrolysis of the acetate derivatives of propargylic alcohols with Bacillus subtilis. However, the optical purities of the products were only up to 74% (Scheme 21.23). 

T. (2006) Fate of transforming bacterial genome following incorporation into competent cells of Bacillus subtilis ... 

Duitman, E.H. et al. (2007) Novel methods for genetic transformation of natural Bacillus subtilis isolates used to study the regulation of the mycosubtilin and surfactin synthetases. Appl. Environ. Microbiol, I i (11),... 

Ohse, M. et al (1995) Effects of plasmid DNA sizes and several other factors on transformation of Bacillus subtilis ISW1214 with plasmid DNA by electroporation. Biosci. BiotechnoL, Biochem., 59 (8), 1433-1437. 

Xue, G.-P., Johnson, J.S, and Dalrymple, B.P. (1999) High osmolarity improves the electro-transformation efficiency of the gram-positive bacteria Bacillus subtilis and Bacillus licheni-formis. J. Microbiol Methods, 34 (3), 183-191. 

Cao, G. et al. (2011) A modified electro-transformation method for Bacillus subtilis and its application in the production of antimicrobial lipopeptides. Biotechnol Lett, 33 (5), 1047-1051. 

Meddeb-Mouelhi, F., Dulcey, C., and Beauregard, M. (2012) High transformation efficiency of Bacillus subtilis... 

Lu, Y.P. et al (2012) Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis. Lett. Appl. Microbiol, 55 (1), 9-14. 

Brigidi, P. et al (1990) Genetic transformation of intact cells of Bacillus subtilis by electroporation. FEMS Microbiol Lett, 67 (1-2), 135-138. 

Akamatsu, T. and Sekiguchi, J. (1987) Characterization of chromosome and plasmid transformation in Bacillus subtilis using gently lysed protoplasts. Arch. Microbiol, 146 (4), 353-357. 

Takeno, M., Taguchi, H., and Akamatsu, T. (2011) Essential involvement of the Bacillus subtilis ABC transporter, EcsB, in genetic transformation of purified DNA but not native DNA from protoplast lysates. /. Biosci. Bioeng., 112 (3), 209-214. 

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