Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2
碩士 === 國立中央大學 === 生命科學系 === 105 === Abstract Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that functions in cell proliferation and differentiation, especially in the growth of white blood cells. In previous studies, the expression of recombinant protein of mGM-CSF (rmGM-C...
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ndltd-TW-105NCU051050162019-05-16T00:08:08Z http://ndltd.ncl.edu.tw/handle/nnbpvb Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2 應用降低表現OsMYBS2水稻轉錄調控因子的策略增加mGM-CSF外源蛋白產量之探討 Desyanti Saulina br. Sinaga 辛娜嘉 碩士 國立中央大學 生命科學系 105 Abstract Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that functions in cell proliferation and differentiation, especially in the growth of white blood cells. In previous studies, the expression of recombinant protein of mGM-CSF (rmGM-CSF) was drove by the αAmy3 promoter in rice suspension cultured cells. The cells were scaled up successfully in a 2-L bioreactor and the highest yield of rmGM-CSF was 24.6 mg/L. Sugar regulates the rice αAmy3 promoter through cis-elements, the GC box, the G box, and the TATCCA element. Three MYB transcription factors OsMYBS1, OsMYBS2, and OsMYBS3, that belong to 1R-MYB, interact specifically with the TATCCA element of αAmy3 promoter and play roles in sugar-regulated αAmy3 expression. The OsMYBS2 is repressor, that is expressed in sugar fed cells, can bind to the TATCCA element, so only a low level of αAmy3 is expressed. To increase the production of rmGM-CSF, the RNAi strategy was used to repress expression of OsMYBS2. The production of the mGM-CSF by aAmy3 promoter-base rice recombinant protein expression system was examined in OsMYBS2 knockdown rice suspension cultured cells. Our results showed that the expression of mGM-CSF gene was higher 1.3 – 5.25 fold under sugar starvation and 1.13 – 4.4 fold in sugar available condition in the OsMYBS2 knockdown cells than wild type. The Western blotting analysis showed that rmGM-CSF protein in OsMYBS2 knockdown was much higher than wild type. In day 5, productioon of rmGM-CSF in OsMYBS2 knockdown cell lines were higher 1.7 – 2.5 fold than in wild-type cell line. The highest yield of recombinant rmGM-CSF was 29.90 µg/mL in 5 days. The high rmGM-CSF productions in OsMYBS2 knockdown cell lines were still able to detect in 11 days of sugar starvation, as compared to in wild type. These studies demonstrated that knockdown of a transcription repressor OsMYBS2 increase the αAmy3 drove mGM-CSF production in sugar starvation of suspension rice cells. Chung-An Lu 陸重安 2017 學位論文 ; thesis 65 zh-TW |
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碩士 === 國立中央大學 === 生命科學系 === 105 === Abstract
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that functions in cell proliferation and differentiation, especially in the growth of white blood cells. In previous studies, the expression of recombinant protein of mGM-CSF (rmGM-CSF) was drove by the αAmy3 promoter in rice suspension cultured cells. The cells were scaled up successfully in a 2-L bioreactor and the highest yield of rmGM-CSF was 24.6 mg/L. Sugar regulates the rice αAmy3 promoter through cis-elements, the GC box, the G box, and the TATCCA element. Three MYB transcription factors OsMYBS1, OsMYBS2, and OsMYBS3, that belong to 1R-MYB, interact specifically with the TATCCA element of αAmy3 promoter and play roles in sugar-regulated αAmy3 expression. The OsMYBS2 is repressor, that is expressed in sugar fed cells, can bind to the TATCCA element, so only a low level of αAmy3 is expressed. To increase the production of rmGM-CSF, the RNAi strategy was used to repress expression of OsMYBS2. The production of the mGM-CSF by aAmy3 promoter-base rice recombinant protein expression system was examined in OsMYBS2 knockdown rice suspension cultured cells. Our results showed that the expression of mGM-CSF gene was higher 1.3 – 5.25 fold under sugar starvation and 1.13 – 4.4 fold in sugar available condition in the OsMYBS2 knockdown cells than wild type. The Western blotting analysis showed that rmGM-CSF protein in OsMYBS2 knockdown was much higher than wild type. In day 5, productioon of rmGM-CSF in OsMYBS2 knockdown cell lines were higher 1.7 – 2.5 fold than in wild-type cell line. The highest yield of recombinant rmGM-CSF was 29.90 µg/mL in 5 days. The high rmGM-CSF productions in OsMYBS2 knockdown cell lines were still able to detect in 11 days of sugar starvation, as compared to in wild type. These studies demonstrated that knockdown of a transcription repressor OsMYBS2 increase the αAmy3 drove mGM-CSF production in sugar starvation of suspension rice cells.
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author2 |
Chung-An Lu |
author_facet |
Chung-An Lu Desyanti Saulina br. Sinaga 辛娜嘉 |
author |
Desyanti Saulina br. Sinaga 辛娜嘉 |
spellingShingle |
Desyanti Saulina br. Sinaga 辛娜嘉 Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2 |
author_sort |
Desyanti Saulina br. Sinaga |
title |
Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2 |
title_short |
Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2 |
title_full |
Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2 |
title_fullStr |
Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2 |
title_full_unstemmed |
Enhanced Production of Granulocyte-Macrophage Colony-Stimulating Factor Production in Rice Suspension Cell by Knockdown of a Transcription Repressor, OsMYBS2 |
title_sort |
enhanced production of granulocyte-macrophage colony-stimulating factor production in rice suspension cell by knockdown of a transcription repressor, osmybs2 |
publishDate |
2017 |
url |
http://ndltd.ncl.edu.tw/handle/nnbpvb |
work_keys_str_mv |
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