{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,13]],"date-time":"2026-09-13T18:02:45Z","timestamp":1789322565498,"version":"build-2803163510"},"reference-count":22,"publisher":"Ovid Technologies (Wolters Kluwer Health)","content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":[],"abstract":"Abstract<\/jats:title>\n \n Background:<\/jats:title>\n \n Interferon-beta (IFN-\u03b2) is a cytokine with a wide range of biological and pharmaceutical applications, including multiple sclerosis (MS), cancer, some autoimmune disorders, and viral infectious diseases. Thus, many studies have been performed to develop novel strategies for the high-yield production of functional IFN-\u03b2 in a cost-effective approach. Here, we aimed to improve the intracellular expression of IFN-\u03b2-1a in\n Pichia pastoris<\/jats:italic>\n .\n <\/jats:p>\n <\/jats:sec>\n \n Materials and Methods:<\/jats:title>\n \n The gene of IFN-\u03b2-1a was successfully sub-cloned into the pPICZA vector. The recombinant vector was transfected to\n P. pastoris<\/jats:italic>\n GS115 cells by electroporation. After screening positive\n P. pastoris<\/jats:italic>\n transformants, the expression of IFN-\u03b2-1a was evaluated and the cultivation conditions, including temperature, time of incubation, and methanol concentration, were optimized. The protein expression levels were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).\n <\/jats:p>\n <\/jats:sec>\n \n Results:<\/jats:title>\n \n The double digestion with\n Eco<\/jats:italic>\n RI and\n Xho<\/jats:italic>\n I restriction enzymes and sequence analysis confirmed the correct sub-cloning of the IFN-\u03b2-1a gene into pPICZA. SDS-PAGE analysis showed that the highest level of IFN-\u03b2-1a (25 mg per 1 L of yeast culture) was produced with 2% methanol at 28\u00b0C after 72 h incubation.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion:<\/jats:title>\n \n Optimization of cultivation conditions for intracellular expression of IFN-\u03b2-1a was successfully performed. This approach can be generally applied to improve the production yield and quality of other recombinant proteins in\n P. pastoris<\/jats:italic>\n .\n <\/jats:p>\n <\/jats:sec>","DOI":"10.4103\/abr.abr_376_23","type":"journal-article","created":{"date-parts":[[2024,8,28]],"date-time":"2024-08-28T04:04:31Z","timestamp":1724817871000},"update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":2,"title":["Cloning and Optimization of Intracellular Expression of Human Interferon \u03b2-1a in Pichia pastoris GS115"],"prefix":"10.4103","volume":"13","author":[{"given":"Diba","family":"Saeidi","sequence":"first","affiliation":[{"name":"Pharmacy Student Research Committee, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara","family":"Saeidi","sequence":"additional","affiliation":[{"name":"Pharmacy Student Research Committee, School of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fatemeh","family":"Moazen","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vajihe","family":"Akbari","sequence":"additional","affiliation":[{"name":"Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"276","published-online":{"date-parts":[[2024,8]]},"reference":[{"key":"R2-20240830","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1038\/sj.cdd.4401850","article-title":"TLR signaling","volume":"13","author":"Kawai","year":"2006","journal-title":"Cell Death 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