DC Field | Value | Language |
---|---|---|
dc.contributor.author | Hwang, SO | ko |
dc.contributor.author | Chung, JY | ko |
dc.contributor.author | Lee, Gyun-Min | ko |
dc.date.accessioned | 2013-03-04T08:21:23Z | - |
dc.date.available | 2013-03-04T08:21:23Z | - |
dc.date.created | 2012-02-06 | - |
dc.date.created | 2012-02-06 | - |
dc.date.issued | 2003-01 | - |
dc.identifier.citation | BIOTECHNOLOGY PROGRESS, v.19, pp.179 - 184 | - |
dc.identifier.issn | 8756-7938 | - |
dc.identifier.uri | http://hdl.handle.net/10203/82152 | - |
dc.description.abstract | In an attempt to increase the specific thrombopoietin (TPO) productivity (q(TPO)) of recombinant Chinese hamster ovary (rCHO) cells (TPO-33), the effect of expression level of ERp57, an isoform of protein disulfide isomerase, on q(TPO) was investigated. To regulate ERp57 expression level, the Tet-Off system was first introduced in TPO-33 cells and stable Tet-Off cells (TPO-33-Tet-Off) were screened by the luciferase assay. The rCHO cells with a doxycycline-regulated ERp57 expression system (TPO-33-ERp57) were obtained by cotransfection of pTRE-ERp57 and pTK-Hyg expression vectors into TPO-33-Tet-Off cells and subsequent screening by Western blot analysis of ERp57 and an enzyme-linked immunosorbent assay of secreted TPO. Western blot analysis showed that ERp57 expression level in TPO-33-ERp57 cells could be regulated tightly by the addition of different concentrations of doxycycline to a culture medium. A doxycycline concentration of 1 mug/mL, which did not influence cell growth and TPO production of TPO-33-Tet-Off cells, was high enough to suppress the ERp57 expression to a basal level. Compared with the basal level, a 1.7-fold increase in ERp57 expression level was obtained in the absence of doxycycline. This increased expression level of ERp57 resulted in a 2.1-fold increase in q(TPO) without growth inhibition, probably as a result of the chaperone-like activity of ERp57 in CHO cells. Taken together, the results obtained here demonstrate that q(TPO) of rCHO cells can be increased by elevating the expression level of ERp57. | - |
dc.language | English | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.subject | PROTEIN DISULFIDE-ISOMERASE | - |
dc.subject | HAMSTER OVARY CELLS | - |
dc.subject | THIOL-DEPENDENT REDUCTASE | - |
dc.subject | PLATELET PRODUCTION | - |
dc.subject | MOLECULAR-CLONING | - |
dc.subject | PHOSPHOLIPASE-C | - |
dc.subject | GLYCINE BETAINE | - |
dc.subject | SECRETION | - |
dc.subject | IDENTIFICATION | - |
dc.subject | OVEREXPRESSION | - |
dc.title | Effect of doxycycline-regulated ERp57 expression on specific thrombopoietin productivity of recombinant CHO cells | - |
dc.type | Article | - |
dc.identifier.wosid | 000180973100026 | - |
dc.identifier.scopusid | 2-s2.0-0037274196 | - |
dc.type.rims | ART | - |
dc.citation.volume | 19 | - |
dc.citation.beginningpage | 179 | - |
dc.citation.endingpage | 184 | - |
dc.citation.publicationname | BIOTECHNOLOGY PROGRESS | - |
dc.contributor.localauthor | Lee, Gyun-Min | - |
dc.contributor.nonIdAuthor | Hwang, SO | - |
dc.contributor.nonIdAuthor | Chung, JY | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordPlus | PROTEIN DISULFIDE-ISOMERASE | - |
dc.subject.keywordPlus | HAMSTER OVARY CELLS | - |
dc.subject.keywordPlus | THIOL-DEPENDENT REDUCTASE | - |
dc.subject.keywordPlus | PLATELET PRODUCTION | - |
dc.subject.keywordPlus | MOLECULAR-CLONING | - |
dc.subject.keywordPlus | PHOSPHOLIPASE-C | - |
dc.subject.keywordPlus | GLYCINE BETAINE | - |
dc.subject.keywordPlus | SECRETION | - |
dc.subject.keywordPlus | IDENTIFICATION | - |
dc.subject.keywordPlus | OVEREXPRESSION | - |
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