{"id":{"repo_id":"nwu-za","oai_identifier":"oai:repository.nwu.ac.za:10394/41465"},"canonical_url":"https://search.dev.ndltd.org/etd/nwu-za/oai:repository.nwu.ac.za:10394/41465","repository":{"repo_id":"nwu-za","name":"North-West University (South Africa)","base_url":"https://repository.nwu.ac.za/server/oai/request"},"display":{"title":"Klonering en uitdrukking van die mensgeen vir interleukin-3 (IL-3)","abstract":"CLONING AND EXPRESSION OF THE HUMAN GENE FOR INTERLEUKIN-3 (IL-3) The hematopo\"ietic growth factor, IL-3, is responsible for the proliferation and differentiation of pluripotent stemcells to mature effector cells in the body. For the formation of mature effector cells, other hematopo\"ietic factors also play a role. The key role of IL-3 during hematopo\"iesis render this factor great therapeutic potential especially when IL-3 is used in combination with other hematopo\"ietic factors. IL-3 is especially successful in the treatment of chemo- and radio-therapy induced secondary hematopo\"ietic failure. The genes for GMCSF, G-CSF and IL-5 have already been successful cloned and expressed in our laboratory. As IL-3 has great therapeutic potential and because of the dependance of the above mentioned factors on IL-3 for greater therapeutic success we decided to clone and expressed the human gene for IL-3 Initially we attempt to isolate the IL-3 gene from cDNA and genomic libraries. For this IL-3 specific oligonucleotides were designed and synthesized to be used as probes for the screening of libraries as well as primers during PCR for the amplification of parts of the IL-3 gene. The amplification products can also be used as probes for the screening of libraries. The oligonucleotides were unsuccessful as probes for the isolation of IL-3 cDNAfrom a cDNA library. The use of oligonucleotides as primers for PCR however was successful and two PCR-products were obtained. The longest PCR-product (500bp) was then used to screen a genomic library. All the isolated potential positive clones each time contained only part of the IL-3 fragment. By means of a synthetic IL-3 cDNA probe it was confirmed that all the isolated clones only contained the third, fourth and fifth exon of the IL-3 gene. The synthetic IL-3 cDNA was expressed in an eukaryotic and a prokaryotic expression system. The IL-3 cDNA was successfully cloned in an.eukaryotic expression vector and IL-3 specific biological activity was shown by a recombinant clone. The IL-3 cDNA was also successfully cloned in an prokaryotic expression vector. Two transformed E.coli cell lines produced unique proteins which correspond with the human IL-3 protein. By means of an IL- 3 protein standard and HPLC-analysis it was found that the unique proteins were indeed the human IL-3 protein. Thus the human gene for IL-3 was successfully cloned and expressed in two expression systems.","abstract_html":"CLONING AND EXPRESSION OF THE HUMAN GENE FOR INTERLEUKIN-3 (IL-3) The hematopo"ietic growth factor, IL-3, is responsible for the proliferation and differentiation of pluripotent stemcells to mature effector cells in the body. For the formation of mature effector cells, other hematopo"ietic factors also play a role. The key role of IL-3 during hematopo"iesis render this factor great therapeutic potential especially when IL-3 is used in combination with other hematopo"ietic factors. IL-3 is especially successful in the treatment of chemo- and radio-therapy induced secondary hematopo"ietic failure. The genes for GMCSF, G-CSF and IL-5 have already been successful cloned and expressed in our laboratory. As IL-3 has great therapeutic potential and because of the dependance of the above mentioned factors on IL-3 for greater therapeutic success we decided to clone and expressed the human gene for IL-3 Initially we attempt to isolate the IL-3 gene from cDNA and genomic libraries. For this IL-3 specific oligonucleotides were designed and synthesized to be used as probes for the screening of libraries as well as primers during PCR for the amplification of parts of the IL-3 gene. The amplification products can also be used as probes for the screening of libraries. The oligonucleotides were unsuccessful as probes for the isolation of IL-3 cDNAfrom a cDNA library. The use of oligonucleotides as primers for PCR however was successful and two PCR-products were obtained. The longest PCR-product (500bp) was then used to screen a genomic library. All the isolated potential positive clones each time contained only part of the IL-3 fragment. By means of a synthetic IL-3 cDNA probe it was confirmed that all the isolated clones only contained the third, fourth and fifth exon of the IL-3 gene. The synthetic IL-3 cDNA was expressed in an eukaryotic and a prokaryotic expression system. The IL-3 cDNA was successfully cloned in an.eukaryotic expression vector and IL-3 specific biological activity was shown by a recombinant clone. The IL-3 cDNA was also successfully cloned in an prokaryotic expression vector. Two transformed E.coli cell lines produced unique proteins which correspond with the human IL-3 protein. By means of an IL- 3 protein standard and HPLC-analysis it was found that the unique proteins were indeed the human IL-3 protein. Thus the human gene for IL-3 was successfully cloned and expressed in two expression systems.","abstract_has_math":false,"creators":["Steyn, Stefanus Johannes"],"institution":"North-West University (South Africa)","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Pretorius, P.J."],"committee_chairs":[],"committee_members":[],"year":1995,"date_issued":"1995","date_published":"1995","updated_at":"2026-07-24T03:33:42Z","subjects":[],"languages":["other"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10394/41465","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Pretorius, P.J."]},{"key":"dc:creator","label":"Author","values":["Steyn, Stefanus Johannes"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2023-05-22T09:09:53Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2023-05-22T09:09:53Z"]},{"key":"dc:date.issued","label":"Date","values":["1995"]},{"key":"dc:publisher","label":"Institution","values":["North-West University (South Africa)"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["other"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10394/41465"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["MSc (Biochemie), North-West University, Potchefstroom Campus"]},{"key":"dc:description.abstract","label":"Abstract","values":["CLONING AND EXPRESSION OF THE HUMAN GENE FOR INTERLEUKIN-3 (IL-3) The hematopo\"ietic growth factor, IL-3, is responsible for the proliferation and differentiation of pluripotent stemcells to mature effector cells in the body. For the formation of mature effector cells, other hematopo\"ietic factors also play a role. The key role of IL-3 during hematopo\"iesis render this factor great therapeutic potential especially when IL-3 is used in combination with other hematopo\"ietic factors. IL-3 is especially successful in the treatment of chemo- and radio-therapy induced secondary hematopo\"ietic failure. The genes for GMCSF, G-CSF and IL-5 have already been successful cloned and expressed in our laboratory. As IL-3 has great therapeutic potential and because of the dependance of the above mentioned factors on IL-3 for greater therapeutic success we decided to clone and expressed the human gene for IL-3 Initially we attempt to isolate the IL-3 gene from cDNA and genomic libraries. For this IL-3 specific oligonucleotides were designed and synthesized to be used as probes for the screening of libraries as well as primers during PCR for the amplification of parts of the IL-3 gene. The amplification products can also be used as probes for the screening of libraries. The oligonucleotides were unsuccessful as probes for the isolation of IL-3 cDNAfrom a cDNA library. The use of oligonucleotides as primers for PCR however was successful and two PCR-products were obtained. The longest PCR-product (500bp) was then used to screen a genomic library. All the isolated potential positive clones each time contained only part of the IL-3 fragment. By means of a synthetic IL-3 cDNA probe it was confirmed that all the isolated clones only contained the third, fourth and fifth exon of the IL-3 gene. The synthetic IL-3 cDNA was expressed in an eukaryotic and a prokaryotic expression system. The IL-3 cDNA was successfully cloned in an.eukaryotic expression vector and IL-3 specific biological activity was shown by a recombinant clone. The IL-3 cDNA was also successfully cloned in an prokaryotic expression vector. Two transformed E.coli cell lines produced unique proteins which correspond with the human IL-3 protein. By means of an IL- 3 protein standard and HPLC-analysis it was found that the unique proteins were indeed the human IL-3 protein. Thus the human gene for IL-3 was successfully cloned and expressed in two expression systems."]},{"key":"dc:title","label":"Title","values":["Klonering en uitdrukking van die mensgeen vir interleukin-3 (IL-3)"]}]}],"canonical_facts":{"dc:contributor.advisor":["Pretorius, P.J."],"dc:creator":["Steyn, Stefanus Johannes"],"dc:date.accessioned":["2023-05-22T09:09:53Z"],"dc:date.available":["2023-05-22T09:09:53Z"],"dc:date.issued":["1995"],"dc:description":["MSc (Biochemie), North-West University, Potchefstroom Campus"],"dc:description.abstract":["CLONING AND EXPRESSION OF THE HUMAN GENE FOR INTERLEUKIN-3 (IL-3) The hematopo\"ietic growth factor, IL-3, is responsible for the proliferation and differentiation of pluripotent stemcells to mature effector cells in the body. For the formation of mature effector cells, other hematopo\"ietic factors also play a role. The key role of IL-3 during hematopo\"iesis render this factor great therapeutic potential especially when IL-3 is used in combination with other hematopo\"ietic factors. IL-3 is especially successful in the treatment of chemo- and radio-therapy induced secondary hematopo\"ietic failure. The genes for GMCSF, G-CSF and IL-5 have already been successful cloned and expressed in our laboratory. As IL-3 has great therapeutic potential and because of the dependance of the above mentioned factors on IL-3 for greater therapeutic success we decided to clone and expressed the human gene for IL-3 Initially we attempt to isolate the IL-3 gene from cDNA and genomic libraries. For this IL-3 specific oligonucleotides were designed and synthesized to be used as probes for the screening of libraries as well as primers during PCR for the amplification of parts of the IL-3 gene. The amplification products can also be used as probes for the screening of libraries. The oligonucleotides were unsuccessful as probes for the isolation of IL-3 cDNAfrom a cDNA library. The use of oligonucleotides as primers for PCR however was successful and two PCR-products were obtained. The longest PCR-product (500bp) was then used to screen a genomic library. All the isolated potential positive clones each time contained only part of the IL-3 fragment. By means of a synthetic IL-3 cDNA probe it was confirmed that all the isolated clones only contained the third, fourth and fifth exon of the IL-3 gene. The synthetic IL-3 cDNA was expressed in an eukaryotic and a prokaryotic expression system. The IL-3 cDNA was successfully cloned in an.eukaryotic expression vector and IL-3 specific biological activity was shown by a recombinant clone. The IL-3 cDNA was also successfully cloned in an prokaryotic expression vector. Two transformed E.coli cell lines produced unique proteins which correspond with the human IL-3 protein. By means of an IL- 3 protein standard and HPLC-analysis it was found that the unique proteins were indeed the human IL-3 protein. Thus the human gene for IL-3 was successfully cloned and expressed in two expression systems."],"dc:identifier.uri":["http://hdl.handle.net/10394/41465"],"dc:language.iso":["other"],"dc:publisher":["North-West University (South Africa)"],"dc:title":["Klonering en uitdrukking van die mensgeen vir interleukin-3 (IL-3)"],"dc:type":["Thesis"]},"updated_at":"2026-07-24T03:33:42Z"}