RUI: Molecular Analysis of the Poly-Beta-Hydroxybutyrate    Biosynthetic Pathway
{'Code': '08070100', 'Directorate': {'Abbreviation': 'BIO', 'LongName': 'Direct For Biological Sciences'}, 'Division': {'Abbreviation': 'MCB', 'LongName': 'Div Of Molecular and Cellular Bioscience'}}
{'SignBlockName': 'Philip Harriman', 'PO_EMAI': None, 'PO_PHON': None}
The goal of this project is to analyze the molecular                            structure of the poly.beta.hydroxybutyrate biosynthetic                         pathway (PHB).  The PHB biosynthetic pathway has been cloned                    from Alcaligenes eutrophus into Escherichia coli where it is                    expressed.  This expression can be induced via oxygen                           limitation or nitrogen limitation.  Proteins encoded by the                     pathway will be specifically translated in vivo and then                        analyzed by denaturing and non.denaturing polyacrylamide gel                    electrophoresis.  Northern blot hybridization and nuclease                      protection assays will be employed to determine the mode of                     transcription and start of transcription, respectively.  The                    three enzymes that constitute the pathway will be initially                     purified utilizing large.scale purification techniques                          followed by fast protein liquid chromatography (FPLC).  Final                   purification will be by preparative isoelectric focusing.  The                  proteins will be characterized as to their size, subunit                        structure, partial amino acid sequence.  Putative PHB                           transcriptional control regions will be located using PHB:lac                   fusions in concert with an induction assay, and agents that                     induce PHB biosynthesis will be identified.                                                                                                                     This is a proposal to lay the groundwork for the molecular and                  genetic study of a metabolic pathway leading to the                             production of a biodegradable polymer in a little studied                       species of soil bacterium.  The bacterium seems to produce this                 polymer as a means of storing energy during times of                            stress, but the mechanism by which it regulates polymer                         production is completely obscure, as are regulatory                             mechanisms in most species of soil bacteria (except for a                       small number which have been well studied).  The work in this                   proposal could lead to the discovery of previously unknown                      mechanisms regulation of gene expression, and is therefore of                   general interest.  It is also of particular interest because of                 what it may tell us about the natural synthesis of the                          biodegradable polymer, information which may have important                     practical consequences.  This work is funded under the RUI                      program, since a great deal of the work is actually performed by                undergraduates who would not otherwise easily be able to                        participate in a serious research program.
{'FirstName': 'Douglas', 'LastName': 'Dennis', 'PI_MID_INIT': 'E', 'PI_SUFX_NAME': None, 'PI_FULL_NAME': 'Douglas E Dennis', 'EmailAddress': 'Douglas.Dennis@asu.edu', 'NSF_ID': '000319287', 'StartDate': '09/01/1988', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'James Madison University', 'CityName': 'HARRISONBURG', 'ZipCode': '228013104', 'PhoneNumber': '5405686872', 'StreetAddress': '800 S MAIN ST', 'StreetAddress2': None, 'CountryName': 'United States', 'StateName': 'Virginia', 'StateCode': 'VA', 'CONGRESSDISTRICT': '06', 'CONGRESS_DISTRICT_ORG': 'VA06', 'ORG_UEI_NUM': 'MVTKSCN6NMH3', 'ORG_LGL_BUS_NAME': 'JAMES MADISON UNIVERSITY', 'ORG_PRNT_UEI_NUM': 'X6KEFGLHSJX7'}
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[{'Code': '1112', 'Text': 'Genetic Mechanisms'}, {'Code': '9120', 'Text': 'CROSS-DIRECTORATE PROGRAMS'}, {'Code': '9145', 'Text': 'SPECIAL PROGRAMS-RESERVE'}]
{'url': 'https://www.nsf.gov/awardsearch/download?DownloadFileName=1988&All=true', 'xml': '8803724.xml'}