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Cytosine deaminase (CodA) is a protein that plays a crucial role in pyrimidine metabolism. It catalyzes the deamination of cytosine and its analog, 5-fluorocytosine (5-FC), to uracil and 5-fluorouracil (5-FU), respectively [1]. This conversion is significant as 5-FU is highly toxic and has been utilized in counterselection strategies due to the ability of CodA to convert the innocuous 5-FC into the toxic 5-FU [2]. The gene encoding CodA has been used as a negative selection marker in various organisms, including plants and bacteria, for genetic modification and mutant strain isolation [3] [4] [5]. Furthermore, CodA has been employed in therapeutic applications, where its expression in bacteria such as Salmonella has led to improved cytotoxic effects in tumor cells [6].
The substrate specificity of CodA is noteworthy, as it can also convert cytosine to uracil, expanding its role beyond 5-FC metabolism [7]. Additionally, the relaxed substrate specificity of CodA allows it to function in pyrimidine metabolism, making it a valuable tool in genetic engineering and metabolic engineering applications [2] [8]. The protein's ability to catalyze the conversion of 5-FC to 5-FU has been exploited in markerless gene deletion techniques, enabling precise manipulation of bacterial chromosomes [1] [3].
References:
[1] L. Wang, J. Hoffmann, H. Watzlawick, & J. Altenbuchner, "Markerless gene deletion with cytosine deaminase in thermus thermophilus strain hb27", Applied and Environmental Microbiology, vol. 82, no. 4, p. 1249-1255, 2016. https://doi.org/10.1128/aem.03524-15
[2] R. Joseph, N. Kim, & N. Sandoval, "Recent developments of the synthetic biology toolkit for clostridium", Frontiers in Microbiology, vol. 9, 2018. https://doi.org/10.3389/fmicb.2018.00154
[3] M. Al-Hinai, A. Fast, & E. Papoutsakis, "Novel system for efficient isolation of clostridium double-crossover allelic exchange mutants enabling markerless chromosomal gene deletions and dna integration", Applied and Environmental Microbiology, vol. 78, no. 22, p. 8112-8121, 2012. https://doi.org/10.1128/aem.02214-12
[4] M. Oliveira, E. Stover, & J. Thomson, "The coda gene as a negative selection marker in citrus", Springerplus, vol. 4, no. 1, 2015. https://doi.org/10.1186/s40064-015-1047-y
[5] M. Shao, J. Michno, S. Hotton, A. Blechl, & J. Thomson, "A bacterial gene coda encoding cytosine deaminase is an effective conditional negative selectable marker in glycine max", Plant Cell Reports, vol. 34, no. 10, p. 1707-1716, 2015. https://doi.org/10.1007/s00299-015-1818-5
[6] B. Mesa-Pereira, C. Medina, E. Camacho, A. Flores, & E. Santero, "Improved cytotoxic effects of salmonella‐producing cytosine deaminase in tumour cells", Microbial Biotechnology, vol. 8, no. 1, p. 169-176, 2014. https://doi.org/10.1111/1751-7915.12153
[7] S. Cartman, M. Kelly, D. Heeg, J. Heap, & N. Minton, "Precise manipulation of the clostridium difficile chromosome reveals a lack of association between the tcdc genotype and toxin production", Applied and Environmental Microbiology, vol. 78, no. 13, p. 4683-4690, 2012. https://doi.org/10.1128/aem.00249-12
[8] R. Wheatley, V. Ramachandran, B. Geddes, B. Perry, C. Yost, & P. Poole, "Role of o2 in the growth of rhizobium leguminosarum bv. viciae 3841 on glucose and succinate", Journal of Bacteriology, vol. 199, no. 1, 2017. https://doi.org/10.1128/jb.00572-16
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