The Influence of DNA Sequences on Recombination in          Mammalian Cells
{'Code': '08070100', 'Directorate': {'Abbreviation': 'BIO', 'LongName': 'Direct For Biological Sciences'}, 'Division': {'Abbreviation': 'MCB', 'LongName': 'Div Of Molecular and Cellular Bioscience'}}
{'SignBlockName': 'DeLill Nasser', 'PO_EMAI': None, 'PO_PHON': None}
Mammalian cells have the ability to carry out homologous                        recombination.  The frequency of recombination in chromosomal                   genes, however,is low and in gene transfer experiments its                      detection is usually obscured by high levels of non-                            homologous recombination.  A variety of plasmid systems have                    now been developed that allow quite detailed studies of                         homologous recombination in cells grown in culture.  The systems                have been further developed assays for recombinational activity                 in extracts prepared from nuclei of such cells can be done. We                  have begun to purify an enzyme activity capable of strand                       exchange between homologous DNA molecules.  It is believed that                 the process of recombination is  essentially similar in all                     organisms.  From studies on  simpler organisms the general                      features of recombination have been determined to be: an                        initiation event, possibly involving preference for specific DNA                sequences; strand exchange between homologous molecules; mismatch               correction ofthe resulting heteroduplex; and finally resolution                 of the synapsed molecules.  We propose to study the enzymology                  and DNA sequence specificity of the first two stages of this                    process, initiation and strand exchange.  Two types of DNA                      sequences, Z DNA and the hypervariable minisatellites, have                     been implicated in promoting recombination in eukaryotes.  We                   have designed a pair of plasmid/phage vectors that can be                       used to assay recombination in the neo gene and allow                           specific sequences to be inserted at homologous sites in each                   substrate to determine their effect on recombination.  Z DNA                    and mini-satellite core repeats will be inserted into these                     vectors and their effects on recombination frequency                            determined by transfection into human EJ cells.  It is                          thought that the presence of such sequences may stimulate the                   initiation and/or pairing  events during recombination.  The                    effects of these sequences on recombination will also be                        examined in the in vitro assay catalyzed by extracts                            prepared from EJ nuclei.                                                                                                                                        The achievement of homologous recombination in mammalian                        cells is of significant interest to geneticists who wish to                     understand basic genetic mechanisms for what insights it                        would give.  Also, the ability to replace genes in their                        correct position in the mammalian genome would have both                        fundamental and commercial importance.
{'FirstName': 'Peter', 'LastName': 'Moore', 'PI_MID_INIT': 'D', 'PI_SUFX_NAME': None, 'PI_FULL_NAME': 'Peter D Moore', 'EmailAddress': None, 'NSF_ID': '000296374', 'StartDate': '07/01/1988', 'EndDate': None, 'RoleCode': 'Principal Investigator'}
{'Name': 'University of Illinois at Chicago', 'CityName': 'CHICAGO', 'ZipCode': '606124305', 'PhoneNumber': '3129962862', 'StreetAddress': '809 S MARSHFIELD AVE M/C 551', 'StreetAddress2': None, 'CountryName': 'United States', 'StateName': 'Illinois', 'StateCode': 'IL', 'CONGRESSDISTRICT': '07', 'CONGRESS_DISTRICT_ORG': 'IL07', 'ORG_UEI_NUM': 'W8XEAJDKMXH3', 'ORG_LGL_BUS_NAME': 'UNIVERSITY OF ILLINOIS', 'ORG_PRNT_UEI_NUM': None}
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{'Code': '111600', 'Text': 'EUKARYOTIC GENETICS'}
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