Autoregulation of Gene Expression by an Allosteric Enzyme
{'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}
Analyses of simple and complex genetic loci in microorganisms                   continue to reveal the presence of new and distinct regulatory                  mechanisms.  The ilv genes of Escherichia coli K-12 will serve                  as the principle focus of the proposed research.                                                                                                                The objectives are:                                                                                                                                               1.  Identify and characterize the sequences required for the                  expression and potential regulation of the promoters located in                 the ilvGMEDA cluster by selecting mutants in vivo with altered                  expression of an indicator gene, use nuclease Bal-31 to                         generate deletions in vitro, and use standard in vitro                          mutagenesis techniques to generate mutants predicted to have                    specific phenotypes.                                                                                                                                              2. Clone the coding sequences of the ilvD and ilvA genes to                   suitable expression vectors, such as plasmid pk30 or pAS1, so                   that these enzymes can be purified for in vitro studies and for                 amino terminal sequence analysis, in order to compare the                       actual and predicted translational start sites for these gene                   products.                                                                                                                                                         3.  Test the apparent dependence of the synthesis of the ilvO                 coded 15 kd protein upon ilvG expression using ilvO-lacZ                        fusions on plasmids and as single copy chromosomal forms.                       Isogenic strains with and without chromosomal deletions of ilvO                 will be monitored for expression and regulation of the ilvGEMDA                 genes.                                                                                                                                                            4.  Test our working hypothesis that the inhibition of cell                   growth observed following arabinose induced expression of ilvO                  on plasmid pMC81 derivatives is due to readthrough into the                     closely linked rrnC cluster and the production of anti-sense                    transcripts of the unique tRNA Trp gene trailer of the rrnC                     cluster.
{'FirstName': 'David', 'LastName': 'Calhoun', 'PI_MID_INIT': 'H', 'PI_SUFX_NAME': None, 'PI_FULL_NAME': 'David H Calhoun', 'EmailAddress': 'calhoun@sci.ccny.cuny.edu', 'NSF_ID': '000352202', 'StartDate': '10/01/1987', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'CUNY City College', 'CityName': 'NEW YORK', 'ZipCode': '100319101', 'PhoneNumber': '2126505418', 'StreetAddress': '160 CONVENT AVE', 'StreetAddress2': None, 'CountryName': 'United States', 'StateName': 'New York', 'StateCode': 'NY', 'CONGRESSDISTRICT': '13', 'CONGRESS_DISTRICT_ORG': 'NY13', 'ORG_UEI_NUM': 'L952KGDMSLV5', 'ORG_LGL_BUS_NAME': 'RESEARCH FOUNDATION OF THE CITY UNIVERSITY OF NEW YORK', 'ORG_PRNT_UEI_NUM': None}
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[{'Code': '111200', 'Text': 'Genetic Mechanisms'}, {'Code': '912000', 'Text': 'CROSS-DIRECTORATE PROGRAMS'}, {'Code': '914500', 'Text': 'SPECIAL PROGRAMS-RESERVE'}]
{'url': 'https://www.nsf.gov/awardsearch/download?DownloadFileName=1987&All=true', 'xml': '8703610.xml'}