Expression, refolding, and characterization of recombinant thrombopoietin/stem cell factor fusion protein in Escherichia coli
文献类型： 外文期刊
作者： Zang, Yuhui 1 ; Zhang, Xu 2 ; Jiang, Xiaoling 2 ; Li, Haoran 2 ; Zhu, Jie 2 ; Zhang, Chi 2 ; Peng, Wei 3 ; Qin, Junchuan;
作者机构： 1.Nanjing Univ, Sch Life Sci, State Key Lab Pharmaceut Biotechnol, Nanjing 210093, Peoples R China
2.Nanjing Univ, Sch Life Sci, State Key Lab Pharmaceut Biotechnol, Nanjing 210093, Peoples R China; Jiangsu Acad Agr Sci, Inst Agrobiol Genet & Physiol, Nanjing 210014, Peoples R China
3.Nanjin
关键词： TPO/SCF;megakaryopoiesis;bacterial expression;inclusion body;refolding;COLONY-STIMULATING FACTOR;GROWTH-RESPONSE;IN-VIVO;INTERLEUKIN-3;HEMATOPOIESIS;LEUKEMIA;IL-3;LINE
期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）
ISSN：
年卷期：
页码：
收录情况： SCI
摘要： Thrombopoietin/stem cell factor (TPO/SCF) is a novel fusion protein that combines the complementary biological effects of TPO and SCF into a single molecule. In this study, TPO/SCF gene was cloned into pET32a and expressed as a thioredoxin (Trx) fusion protein with a C-terminal 6His-tag in Escherichia coli BL21(DE3) under the control of T7 promoter. Trx-TPO/SCF protein approximately accounted for 20% of the total bacterial proteins and was found to accumulate in inclusion bodies. Inclusion bodies were separated from cellular debris, washed with buffer containing 2 M urea, and solubilized with 8 M urea. The refolding of Trx-TPO/SCF was then carried out by an on-column method. Soluble Trx-TPO/SCF was characterized for its dose-dependent effects on promoting cells proliferation in both TF1 and Mo7e cell lines. rhTPO/SCF was released by thrombin digestion and further purified by Ni2+ affinity chromatography. Western blot analysis confirmed the identities of Trx-TPO/SCF and rhTPO/SCF.
- 相关文献
作者其他论文 更多>>
-
A phase-separated protein hub modulates resistance to Fusarium head blight in wheat
作者：He, Yi;Wang, Yuhua;Dong, Yifan;Zhang, Zhengguang;Li, Gang;He, Yi;Wu, Lei;Jiang, Peng;Zhang, Xu;Yang, Xiujuan;Tucker, Matthew R.;Xia, Xiaobo;Jiang, Cong;Ma, Hongxiang;Ma, Wujun;Liu, Cong;Whitford, Ryan
关键词：
-
A novel approach for estimating fractional cover of crops by correcting angular effect using radiative transfer models and UAV multi-angular spectral data
作者：Pan, Yuanyuan;Wu, Wenxuan;He, Jiaoyang;Zhu, Jie;Su, Xi;Li, Wanyu;Li, Dong;Yao, Xia;Cheng, Tao;Zhu, Yan;Cao, Weixing;Tian, Yongchao;Pan, Yuanyuan
关键词：Multi-FVC model; Multi-angle remote sensing; Soil background, PROSAIL; Gaussian process regression
-
Large-scale breeding population validating significant loci for the 1000-kernel weight of wheat
作者：Jiang, Peng;Li, Chang;Wu, Lei;He, Yi;Zhang, Peng;Zhang, Xu;Jiang, Peng;Wu, Lei;He, Yi;Zhang, Peng;Zhang, Xu;Cui, Fa;Zhang, Zhaoyang;Ma, Hongxiang;Zhang, Xu;Jiang, Peng;Zhang, Xu
关键词：
-
Cost-effective duplex Kompetitive Allele Specific PCR markers for homologous genes facilitating wheat breeding
作者：Jiang, Peng;Fan, Xiangyun;Wu, Lei;He, Yi;Li, Chang;Zhang, Xu;Zhang, Guangxu;Zhang, Xu
关键词：Favorable allele; Kompetitive Allele Specific PCR; Marker-assisted selection; Wheat breeding
-
Comparative transcriptomics identifies the key in planta-expressed genes of Fusarium graminearum during infection of wheat varieties
作者：Tu, Qiang;Wang, Lirong;An, Qi;Shuai, Jie;Dong, Yifan;Zhang, Xu;He, Yi;Tu, Qiang;Wang, Lirong;An, Qi;Shuai, Jie;Xia, Xiaobo;Li, Gang;He, Yi
关键词：transcriptome; gene expression; Fusarium head blight; resistance; susceptibility; F; graminearum; wheat
-
Investigating the Resistance Mechanism of Wheat Varieties to Fusarium Head Blight Using Comparative Metabolomics
作者：Dong, Yifan;Ahmad, Dawood;Zhang, Xu;Wu, Lei;Jiang, Peng;Zhang, Peng;He, Yi;Yang, Xiujuan;Xia, Xiaobo;Wang, Yuhua;Li, Gang;Ahmad, Dawood
关键词：metabolomics; wheat; Fusarium head blight; fungistatic; gene expression; resistance; susceptibility
-
Identification of adult plant yellow rust resistance QTLs in Jiangsu wheat varieties Ningmai 9 and Yangmai 158
作者：Jiang, Peng;Wu, Lei;He, Yi;Li, Chang;Zhang, Peng;Zhang, Xu;Wu, Ling;Ren, Yong;Zhang, Hua;Ma, Hongxiang;Zhang, Xu
关键词：linkage mapping; marker-assisted selection; wheat breeding; yellow rust
