Difference between revisions of "Part:BBa K318512:Experience"

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Applications of BBa_K318512

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UNIQ325dbe3d57924a3b-partinfo-00000000-QINU UNIQ325dbe3d57924a3b-partinfo-00000001-QINU

gadAp: A pH Sensitive Promoter. One of the most apparent environmental changes which occurs upon chyme movement from the stomach to the small intestine is a sharp (approximately 2-10) pH gradient. However, in order to utilize this as a signaling mechanism for cell lysis, we required a pH sensitive element. Results of northern blot analysis (Tucker et al., 2002; Waterman et al., 2003) indicate that a particular glutamate decarboxylase (GadA) is up-regulated, most likely as a part of a glutamate to gamma-aminobutyrate reaction which exports intercelluar protons which is required for E. coli stationary phase cell survival of highly acidic (pH 2) conditions (Castanie-Cornet et al., 1999). Based on a high ratio of low pH:neutral pH expression and a wealth of available information, we chose to use GadA's promoter, gadAp, as a native-E. coli sensor for acidic conditions.

General information about this promoter and other related acid-sensitive promoters in this system can be found here: http://biocyc.org/ECOLI/NEW-IMAGE?type=OPERON-IN-CHROM-BROWSER&object=TU586 Ecocyc-gadAp.

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-'''gadAp: A pH Sensitive Promoter'''. One of the most apparent environmental changes which occurs upon chyme movement from the stomach to the small intestine is a sharp (approximately 2-10)  pH gradient. However, in order to utilize this as a signaling mechanism for cell lysis, we required a pH sensitive element. Results of northern blot analysis (Tucker ''et al.'', 2002; Waterman ''et al.'', 2003) indicate that a particular glutamate decarboxylase (GadA) is up-regulated, most likely as a part of a glutamate to gamma-aminobutyrate reaction which exports intercelluar protons which is required for ''E. coli'' stationary phase cell survival of highly acidic (pH 2) conditions (Castanie-Cornet ''et al.'', 1999). Based on a high ratio of low pH:neutral pH expression and a wealth of available information, we chose to use GadA's promoter, gadAp, as a native-''E. coli'' sensor for acidic conditions. In order to maximize pH-mediated response of the gadA promoter and minimize induction caused by other known responses (such as the stationary phase), we plan to run error-prone PCR followed by screening of clones for maximal, dedicated acid response.
+'''gadAp: A pH Sensitive Promoter'''. One of the most apparent environmental changes which occurs upon chyme movement from the stomach to the small intestine is a sharp (approximately 2-10)  pH gradient. However, in order to utilize this as a signaling mechanism for cell lysis, we required a pH sensitive element. Results of northern blot analysis (Tucker ''et al.'', 2002; Waterman ''et al.'', 2003) indicate that a particular glutamate decarboxylase (GadA) is up-regulated, most likely as a part of a glutamate to gamma-aminobutyrate reaction which exports intercelluar protons which is required for ''E. coli'' stationary phase cell survival of highly acidic (pH 2) conditions (Castanie-Cornet ''et al.'', 1999). Based on a high ratio of low pH:neutral pH expression and a wealth of available information, we chose to use GadA's promoter, gadAp, as a native-''E. coli'' sensor for acidic conditions.
 General information about this promoter and other related acid-sensitive promoters in this system can be found here: [[http://biocyc.org/ECOLI/NEW-IMAGE?type=OPERON-IN-CHROM-BROWSER&object=TU586 Ecocyc-gadAp]].