Soluble expression of recombinant cyclophilin A in Escherichia coli
To produce a large amount of soluble cyclophilin A (CypA) protein, the recombinant pRSET -CypA plasmid was transferred into E. coli BL21. Using the 2×YT culture medium, the medium compositions of carbon source, nitrogen source and ampicillin concentration were optimized, and the effects of inoculum...
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Zhejiang University Press
2010-07-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.04.002 |
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| author | YAN Xiao XU Li-ren GUAN Yi-xin YAO Shan-jing |
| author_facet | YAN Xiao XU Li-ren GUAN Yi-xin YAO Shan-jing |
| author_sort | YAN Xiao |
| collection | DOAJ |
| description | To produce a large amount of soluble cyclophilin A (CypA) protein, the recombinant pRSET -CypA plasmid was transferred into E. coli BL21. Using the 2×YT culture medium, the medium compositions of carbon source, nitrogen source and ampicillin concentration were optimized, and the effects of inoculum and induction conditions including inducer concentration, induction time and incubation time were investigated. The results indicated that the recombinant CypA expressed as inclusion bodies was greatly inhibited by controlling the culture temperature and rotating speed of incubator. The mannitol as carbon source could increase the CypA production effectively. The optimized culture medium was composed of peptone 16 g·L<sup>-1</sup>, yeast extract 10 g·L<sup>-1</sup>, NaCl 5 g·L<sup>-1</sup>, mannitol 10 g·L<sup>-1</sup>, Amp 50 mg·L<sup>-1</sup>. When OD<sub>600</sub> was 1.3, IPTG of 1 mmol·L<sup>-1</sup> was added, and the cells were harvested after 9 h cultivation at 30 ℃, 180 r·min<sup>-1</sup>. After the optimization, the soluble CypA expression reached 66.7 mg·L<sup>-1</sup>, increased by 76.6% compared with the level under the preoptimized conditions. |
| format | Article |
| id | doaj-art-ec9d7582789442e7b027e873e8e7338f |
| institution | DOAJ |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2010-07-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-ec9d7582789442e7b027e873e8e7338f2025-08-20T03:16:08ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552010-07-013636336910.3785/j.issn.1008-9209.2010.04.00210089209Soluble expression of recombinant cyclophilin A in Escherichia coliYAN XiaoXU Li-renGUAN Yi-xinYAO Shan-jingTo produce a large amount of soluble cyclophilin A (CypA) protein, the recombinant pRSET -CypA plasmid was transferred into E. coli BL21. Using the 2×YT culture medium, the medium compositions of carbon source, nitrogen source and ampicillin concentration were optimized, and the effects of inoculum and induction conditions including inducer concentration, induction time and incubation time were investigated. The results indicated that the recombinant CypA expressed as inclusion bodies was greatly inhibited by controlling the culture temperature and rotating speed of incubator. The mannitol as carbon source could increase the CypA production effectively. The optimized culture medium was composed of peptone 16 g·L<sup>-1</sup>, yeast extract 10 g·L<sup>-1</sup>, NaCl 5 g·L<sup>-1</sup>, mannitol 10 g·L<sup>-1</sup>, Amp 50 mg·L<sup>-1</sup>. When OD<sub>600</sub> was 1.3, IPTG of 1 mmol·L<sup>-1</sup> was added, and the cells were harvested after 9 h cultivation at 30 ℃, 180 r·min<sup>-1</sup>. After the optimization, the soluble CypA expression reached 66.7 mg·L<sup>-1</sup>, increased by 76.6% compared with the level under the preoptimized conditions.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.04.002cyclophilin Asoluble expressionculture optimization |
| spellingShingle | YAN Xiao XU Li-ren GUAN Yi-xin YAO Shan-jing Soluble expression of recombinant cyclophilin A in Escherichia coli 浙江大学学报. 农业与生命科学版 cyclophilin A soluble expression culture optimization |
| title | Soluble expression of recombinant cyclophilin A in Escherichia coli |
| title_full | Soluble expression of recombinant cyclophilin A in Escherichia coli |
| title_fullStr | Soluble expression of recombinant cyclophilin A in Escherichia coli |
| title_full_unstemmed | Soluble expression of recombinant cyclophilin A in Escherichia coli |
| title_short | Soluble expression of recombinant cyclophilin A in Escherichia coli |
| title_sort | soluble expression of recombinant cyclophilin a in escherichia coli |
| topic | cyclophilin A soluble expression culture optimization |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2010.04.002 |
| work_keys_str_mv | AT yanxiao solubleexpressionofrecombinantcyclophilinainescherichiacoli AT xuliren solubleexpressionofrecombinantcyclophilinainescherichiacoli AT guanyixin solubleexpressionofrecombinantcyclophilinainescherichiacoli AT yaoshanjing solubleexpressionofrecombinantcyclophilinainescherichiacoli |