{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/22183"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/22183","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Modification of bovine beta-casein using genetic engineering and its structure/function studies","abstract":"Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:55:53Z 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 2011-05-07T14:55:53Z Item is restricted indefinitely.","abstract_has_math":false,"creators":["Choi, Byung-Kwon"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Food Science and Human Nutrition","degree_department":null,"school":null,"contributors":["Jimenez-Flores, Rafael"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T13:31:40Z","date_published":"2011-05-07T13:31:40Z","updated_at":"2026-07-22T22:25:19Z","subjects":["Biology, Molecular","Agriculture, Food Science and Technology","Health Sciences, Nutrition"],"languages":["eng"],"rights":["Copyright 1996 Choi, Byung-Kwon"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9625122","(UMI)AAI9625122"],"render_values":[{"text":"AAI9625122","href":null,"code":true},{"text":"(UMI)AAI9625122","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/22183","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Jimenez-Flores, Rafael"]},{"key":"dc:creator","label":"Author","values":["Choi, Byung-Kwon"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T13:31:40Z","10000-01-01","1996"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Food Science and Human Nutrition"]},{"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","Health Sciences, Nutrition"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 1996 Choi, Byung-Kwon"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9625122","(UMI)AAI9625122","http://hdl.handle.net/2142/22183"]}]},{"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 2011-05-07T14:55:53Z Item is restricted indefinitely.","The putative glycosylation site (Asn-X-Ser) was introduced to the bovine $\\beta$-casein cDNA and genomic DNA (A$\\sp2$ genetic variant) using PCR-based site-directed mutagenesis to increase its amphiphilicity. The two mutant bovine $\\beta$-casein sequences containing the substitution of ${\\bf Ser}\\sb{70}$ for ${\\bf Leu}\\sb{70}$ (Asn$\\sb{68}$-Ser$\\sb{69}$-${\\bf Ser}\\sb{70}$-Pro$\\sb{71})$ and the substitution of ${\\bf Ser}\\sb{71}$ for ${\\bf Pro}\\sb{71}$ (Asn$\\sb68$-Ser$\\sb{69}$-${\\bf Ser}\\sb{70}$-${\\bf Ser}\\sb{71})$ were constructed and expressed using Pichia pastoris and transgenic mice. The mutant constructs were placed under the control of the methanol-inducible alcohol oxidase (AOX1) promoter for P. pastoris and of the bovine $\\alpha$-lactalbumin promoter for transgenic mice. The N-linked glycosylation of the mutant carrying Pro$\\sb{71}$ was completely inhibited whereas the other mutant carrying Ser$\\sb{71}$ was N-linked glycosylated in both expression systems. Thus, the C-terminal proline residue in the sequon was found to be a structural inhibitor for N-glycosylation. Glyco-$\\beta$-caseins produced in both systems were phosphorylated to the same degree as authentic bovine $\\beta$-casein. In addition, based on lectin blots, the oligosaccharides of glyco-$\\beta$-casein from P. pastoris were high-mannose type whereas transgenic glyco-$\\beta$-caseins contained various oligosaccharide chains. Mouse casein micelles containing glycosylated bovine $\\beta$-casein showed the largest median diameter and rough outer surfaces, compared to normal mouse casein micelles and micelles from transgenic milk containing wild-type bovine $\\beta$-casein.","Made available in DSpace on 2011-05-07T13:31:40Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9625122.pdf: 5991212 bytes, checksum: c629897571643e48626e39e336256290 (MD5) Previous issue date: 1996","Restriction data tranferred 2014-07-01T11:26:05-05:00 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":["Modification of bovine beta-casein using genetic engineering and its structure/function studies"]}]}],"canonical_facts":{"dc:contributor":["Jimenez-Flores, Rafael"],"dc:creator":["Choi, Byung-Kwon"],"dc:date":["2011-05-07T13:31:40Z","10000-01-01","1996"],"dc:description":["Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:55:53Z Item is restricted indefinitely.","The putative glycosylation site (Asn-X-Ser) was introduced to the bovine $\\beta$-casein cDNA and genomic DNA (A$\\sp2$ genetic variant) using PCR-based site-directed mutagenesis to increase its amphiphilicity. The two mutant bovine $\\beta$-casein sequences containing the substitution of ${\\bf Ser}\\sb{70}$ for ${\\bf Leu}\\sb{70}$ (Asn$\\sb{68}$-Ser$\\sb{69}$-${\\bf Ser}\\sb{70}$-Pro$\\sb{71})$ and the substitution of ${\\bf Ser}\\sb{71}$ for ${\\bf Pro}\\sb{71}$ (Asn$\\sb68$-Ser$\\sb{69}$-${\\bf Ser}\\sb{70}$-${\\bf Ser}\\sb{71})$ were constructed and expressed using Pichia pastoris and transgenic mice. The mutant constructs were placed under the control of the methanol-inducible alcohol oxidase (AOX1) promoter for P. pastoris and of the bovine $\\alpha$-lactalbumin promoter for transgenic mice. The N-linked glycosylation of the mutant carrying Pro$\\sb{71}$ was completely inhibited whereas the other mutant carrying Ser$\\sb{71}$ was N-linked glycosylated in both expression systems. Thus, the C-terminal proline residue in the sequon was found to be a structural inhibitor for N-glycosylation. Glyco-$\\beta$-caseins produced in both systems were phosphorylated to the same degree as authentic bovine $\\beta$-casein. In addition, based on lectin blots, the oligosaccharides of glyco-$\\beta$-casein from P. pastoris were high-mannose type whereas transgenic glyco-$\\beta$-caseins contained various oligosaccharide chains. Mouse casein micelles containing glycosylated bovine $\\beta$-casein showed the largest median diameter and rough outer surfaces, compared to normal mouse casein micelles and micelles from transgenic milk containing wild-type bovine $\\beta$-casein.","Made available in DSpace on 2011-05-07T13:31:40Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9625122.pdf: 5991212 bytes, checksum: c629897571643e48626e39e336256290 (MD5) Previous issue date: 1996","Restriction data tranferred 2014-07-01T11:26:05-05:00 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":["AAI9625122","(UMI)AAI9625122","http://hdl.handle.net/2142/22183"],"dc:language":["eng"],"dc:rights":["Copyright 1996 Choi, Byung-Kwon"],"dc:subject":["Biology, Molecular","Agriculture, Food Science and Technology","Health Sciences, Nutrition"],"dc:title":["Modification of bovine beta-casein using genetic engineering and its structure/function studies"],"dc:type":["text"],"thesis:degree_discipline":["Food Science and Human Nutrition"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:25:19Z"}