{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/19335"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/19335","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Genetic construction of starch-utilizing strain of Saccharomyces cerevisiae and stabilization of its recombinant plasmid by immobilization","abstract":"Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:36:15Z Item is restricted indefinitely.","abstract_html":"Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:36:15Z Item is restricted indefinitely.","abstract_has_math":false,"creators":["Kim, Chung Gyu"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Food Science","degree_department":null,"school":null,"contributors":["Witter, Lloyd D."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T12:04:19Z","date_published":"2011-05-07T12:04:19Z","updated_at":"2026-07-22T22:25:12Z","subjects":["Biology, Molecular","Agriculture, Food Science and Technology","Biology, Microbiology"],"languages":["eng"],"rights":["Copyright 1990 Kim, Chung Gyu"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9114292","(UMI)AAI9114292"],"render_values":[{"text":"AAI9114292","href":null,"code":true},{"text":"(UMI)AAI9114292","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/19335","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Witter, Lloyd D."]},{"key":"dc:creator","label":"Author","values":["Kim, Chung Gyu"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T12:04:19Z","10000-01-01","1990"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Food Science"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biology, Molecular","Agriculture, Food Science and Technology","Biology, Microbiology"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 1990 Kim, Chung Gyu"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9114292","(UMI)AAI9114292","http://hdl.handle.net/2142/19335"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:36:15Z Item is restricted indefinitely.","DNA containing the nucleotide sequence for $\\alpha$-amylase activity has been isolated from Bacillus stearothermophilus and shot-gun cloned into E. coli host using a plasmid vector pUC19 by electroporation-induced transformation. The hybrid plasmid (designated as pCK101) containing the amylase gene, after purification of the gene by subcloning (i.e., the trimming and manipulation of the amy$\\sp{+}$ fragment from pUC101), was transformed from the host organism to S. cerevisiae using E. coli/yeast shuttle vector, pMF$\\alpha$8 by electroporation transformation procedure. When the purified B. stearothermophilus $\\alpha$-amylase gene was fused to the pMF$\\alpha$8 just after S. cerevisiae MF$\\alpha$1 promoter and secretion signals coding sequence, the transformants of S. cerevisiae cells were successfully able to synthesize and secrete functional $\\alpha$-amylase efficiently hydrolyzing starch present in the culture medium. Further studies on the characterization of cloned gene was done.","When plasmid stability was studied in continuous air bubble bioreactor, there was a generally observed loss of plasmid with time. The plasmid containing whole cells of S. cerevisiae was immobilized in k-carrageenan and allowed to react for varying period time. Cells were then released from immobilization and examined for plasmid stability, cell mass production and $\\alpha$-amylase production. The transformed S. cerevisiae was then compared for free and immobilized system with and without selection pressure. Data showed that the immobilization of cells carrying recombinant plasmid could improve the plasmid stability. The significantly higher cell concentration and consequently higher productivity of the plasmid-coded $\\alpha$-amylase product was also obtained in the comparison to free cell system.","This research project was initiated in the hope of developing a heterogeneous gene expression and secretion system in S. cerevisiae by choosing the enzyme $\\alpha$-amylase as a model system. Also, it was great concern to associate the methods of both genetic and immobilization engineering in this project. Those results showed good prospectives on its practical use of immobilization technique as a new strategy for maintaining a stable population of plasmid-carrying S. cerevisiae.","Made available in DSpace on 2011-05-07T12:04:19Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9114292.pdf: 5604610 bytes, checksum: cc4a7fae96c02ab4e12f03823e753a21 (MD5) Previous issue date: 1990","Restriction data tranferred 2014-07-01T11:14:35-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"]},{"key":"dc:title","label":"Title","values":["Genetic construction of starch-utilizing strain of Saccharomyces cerevisiae and stabilization of its recombinant plasmid by immobilization"]}]}],"canonical_facts":{"dc:contributor":["Witter, Lloyd D."],"dc:creator":["Kim, Chung Gyu"],"dc:date":["2011-05-07T12:04:19Z","10000-01-01","1990"],"dc:description":["Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:36:15Z Item is restricted indefinitely.","DNA containing the nucleotide sequence for $\\alpha$-amylase activity has been isolated from Bacillus stearothermophilus and shot-gun cloned into E. coli host using a plasmid vector pUC19 by electroporation-induced transformation. The hybrid plasmid (designated as pCK101) containing the amylase gene, after purification of the gene by subcloning (i.e., the trimming and manipulation of the amy$\\sp{+}$ fragment from pUC101), was transformed from the host organism to S. cerevisiae using E. coli/yeast shuttle vector, pMF$\\alpha$8 by electroporation transformation procedure. When the purified B. stearothermophilus $\\alpha$-amylase gene was fused to the pMF$\\alpha$8 just after S. cerevisiae MF$\\alpha$1 promoter and secretion signals coding sequence, the transformants of S. cerevisiae cells were successfully able to synthesize and secrete functional $\\alpha$-amylase efficiently hydrolyzing starch present in the culture medium. Further studies on the characterization of cloned gene was done.","When plasmid stability was studied in continuous air bubble bioreactor, there was a generally observed loss of plasmid with time. The plasmid containing whole cells of S. cerevisiae was immobilized in k-carrageenan and allowed to react for varying period time. Cells were then released from immobilization and examined for plasmid stability, cell mass production and $\\alpha$-amylase production. The transformed S. cerevisiae was then compared for free and immobilized system with and without selection pressure. Data showed that the immobilization of cells carrying recombinant plasmid could improve the plasmid stability. The significantly higher cell concentration and consequently higher productivity of the plasmid-coded $\\alpha$-amylase product was also obtained in the comparison to free cell system.","This research project was initiated in the hope of developing a heterogeneous gene expression and secretion system in S. cerevisiae by choosing the enzyme $\\alpha$-amylase as a model system. Also, it was great concern to associate the methods of both genetic and immobilization engineering in this project. Those results showed good prospectives on its practical use of immobilization technique as a new strategy for maintaining a stable population of plasmid-carrying S. cerevisiae.","Made available in DSpace on 2011-05-07T12:04:19Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9114292.pdf: 5604610 bytes, checksum: cc4a7fae96c02ab4e12f03823e753a21 (MD5) Previous issue date: 1990","Restriction data tranferred 2014-07-01T11:14:35-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"],"dc:identifier":["AAI9114292","(UMI)AAI9114292","http://hdl.handle.net/2142/19335"],"dc:language":["eng"],"dc:rights":["Copyright 1990 Kim, Chung Gyu"],"dc:subject":["Biology, Molecular","Agriculture, Food Science and Technology","Biology, Microbiology"],"dc:title":["Genetic construction of starch-utilizing strain of Saccharomyces cerevisiae and stabilization of its recombinant plasmid by immobilization"],"dc:type":["text"],"thesis:degree_discipline":["Food Science"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:25:12Z"}