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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

[1]Expression, purification, and characterization of a novel recombinant fusion protein, rhTPO/SCF, in Escherichia coli. Zang, YH,Zhang, X,Yuan, DW,Zhang, YM,Zhu, J,Lu, HQ,Chang, C,Qin, JC.

[2]Effects of microcystins on the growth and the activity of superoxide dismutase and peroxidase of rape (Brassica napus L.) and rice (Oryza sativa L.). Chen, JZ,Song, LR,Dai, E,Gan, NQ,Liu, ZL.

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S.;Thapa, Monish Kumar;Javid, Osman;Nagar, Padamnabhi Shanker;Rajan, Priyadarsanan Dharma;Salini, S.;Rathod, Sachin G.;Molur, Sanjay;Singh, Sarvesh Kumar;Phartyal, Shyam S.;Karuppusamy, Subbiah;Sheikh, Taslima;Irham, Mohammad;Muhaimin, Muhamad;Mustaqim, Wendy Achmmad;Yonvitner;Mahdavi, Ahmad;Azani, Nasim;Pourghorban, Zohreh;Zeraatkar, Amin;Zeraatkar, Amin;Pahlevani, Amir Hossein;Pirani, Atefeh;Memarian, Farshid;Moazzeni, Hamid;Habibi, Meysam;Doostmohammadi, Moslem;Griffin, Bogna;Guiry, Michael;Jackson, Patrick N. Wyse;Bouskila, Amos;Shacham, Boaz;Gossa, Tegenu;Ballerio, Alberto;Allievi, Alessandro;de Jager, Gerhard;Petroni, Giulio;Peruzzi, Lorenzo;D'Antraccoli, Marco;Serra, Valentina;Bisi, Alessandro;Chiarucci, Alessandro;Hilpold, Andreas;Rosso, Antonietta;Sciuto, Francesco;Mingozzi, Antonio;Giovanni, Boano;Massa, Bruno;Marrone, Federico;Vecchioni, Luca;Guarino, Riccardo;Giusto, Carlo;Lombardi, Chiara;Nepi, Chiara;Scaccini, Davide;Fontaneto, Diego;Banfi, Enrico;Rigato, Fabrizio;Galasso, Gabriele;Chiozzi, Giorgio;Gallo, Enrico;Bertasi, Fabio;Bartolucci, Fabrizio;Fabbriciani, Fabrizio;Andreone, Franco;Polgar, Gianluca;Filibeck, Goffredo;Tessa, Giulia;Gardini, Giulio;Fenu, Giuseppe;Mazzini, Ilaria;Langeneck, Joachim;Bartolozzi, Luca;Bologna, Marco A.;Abeli, Thomas;Dellacasa, Marco;Uliana, Marco;Novarini, Nicola;Faccoli, Massimo;Majorano, Paolo;Zilioli, Michele;Raia, Pasquale;Cerretti, Pierfilippo;Poggi, Roberto;Bertolino, Sandro;Orsenigo, Simone;Martellos, Stefano;Greuter, Werner;Greuter, Werner;Kouakou, Lombart Maurice;Kato, Daichi;Yadav, Harsh;Chiba, Hideyuki;Chiba, Hideyuki;Yoshitomi, Hiroyuki;Kakui, Keiichi;Kwak, Mackenzie L.;Dick, Matthew H.;Ikuno, Kenji;Genka, Masaaki;Yano, Okihito;Kakizoe, Showtaro;Hellemans, Simon;Jashenko, Roman;Dujsebayeva, Tatjana;Kabak, Ilya;Kabak, Ilya;Katumo, Daniel Mutavi;Baasanmunkh, Shukherdorj;Choi, Hyeok Jae;Shin, Jae-Seo;Eun, Lim Ji;Jung, Eui-Kwon;Kim, Young-Dong;Lazkov, Georgii;Strijk, Joeri Sergej;Shavrin, Alexey;Greke, Kristine;Kalnins, Martins;Petrasiunas, Andrius;Uogintas, Domas;Stonis, Jonas R.;Gudzinskas, Zigmantas;Sokolova, Elena;Kharchenko, Viktoria;Andriambololonera, Sylvie;Kumaran, Jayaraj Vijaya;Appalasamy, Suganthi;Prozorov, Alexey M.;Dembele, Jacques;Hanan Alipi, Ana Maria;Solano Gomez, Aniceto Rodolfo;Maruri-Aguilar, Beatriz;Ramirez Diaz, Claudia Janeth;Carnevali Fernandez-Concha, German;Ramirez-Morillo, Ivon M.;Stefano Duno, Rodrigo;Delgadillo-Moya, Claudio;Prieto Torres, David Alexander;Daniel Tejero-Diez, J.;Meave, Jorge A.;Alvarado Cardenas, Leonardo Osvaldo;Terrazas, Teresa;Barradas, Victor L.;Adilson Pinedo-Escatel, J.;Murugan, Gopal;Ruiz Sanchez, Eduardo;Sanchez Martinez, Emiliano;Hernandez Martinez, Maria Magdalena;Gomez-Noguez, Felipe;Montiel Canales, Gustavo;Socorro Gonzalez-Elizondo, M.;Gonzalez Elizondo, Martha;Rolando Bastida-Zavala, J.;del Socorro Garcia-Madrigal, Maria;Alvarado Rodriguez, Jose Fernando;Lopez Mata, Lauro;Gonzalez Ledesma, Manuel;Ramirez-Marcial, Neptali;Cuevas Guzman, Ramon;Martinez-Camilo, Rub'n;Adilson Pinedo-Escatel, J.;Boldgiv, Bazartseren;Mathieu, Chambouleyron;Lermmel, Claude;Leger, Jean-Francois;Farooq, Harith;Irish, John;Shrestha, Krishna;Dop, A. 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Luisa;Vicens Fornes, Magdalena;Sanchez-Angulo, Manuel;Escudero, Marcial;Alcolea, Marta;Sanchez Gomez, Pedro;Fraga i Arguimbau, Pere;Orihuela-Rivero, Raol;Granjel, Rodrigo R.;Molino, Sonia;Garcia Mendizabal, Yoana;Carlsson, Bengt;Svensson, Brita;Bosagna, Carlos Guerrero;Arnqvist, Goran;Cardenas, Paco;Ahlberg, Per;Alstrom, Per;Liu, Zongzhuang;Alonso, German Hernandez;Wahlsteen, Eric;Bisang, Irene;Jonsson, Knud Andreas;Hedenas, Lars;Wedin, Mats;Noren, Michael;Stohr, Sabine;Kiel, Steffen;Rota, Jadranka;Wahlberg, Niklas;Tyler, Torbjorn;Arup, Ulf;Strid, Arne;Liljeblad, Johan;Lundin, Kennet;Nielsen, Peter;Strid, Arne;Alonso, German Hernandez;Chautems, Alain;Charpin, Andre;Mombrial, Florian;Zamora, Juan Carlos;Fumeaux, Nicolas;Clerc, Philippe;Garcia-Martin, Ana B.;Mueller, Andreas;Freitag, Anne;Landry, Bernard;Cuccodoro, Giulio;Sabatinelli, Guido;Lobl, Ivan;Valtotton, Laurent;Ruedi, Manuel;Schuchert, Peter;Praz, Christophe;Thivaiou, Danae;Klopfstein, Seraina;Cherix, Daniel;Mitchell, Edward;Mariaux, Jean;Haenni, Jean-Paul;Theurillat, Jean-Paul;Theurillat, Jean-Paul;Bernasconi, Marvco V.;Hawlitschek, Oliver;Samathi, Adun;Hyde, Kevin;Radji, Raoufou;Morales, J. 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V., Jr.;Esselstyn, Jacob;Eberhard, Jessica;Harms, Kyle E.;Johnson, Andrew;Capparella, Angelo;Rossi, Anthony;Casamatta, Dale;Namayandeh, Armin;Borkent, Art;Smith, Brian Tilston;Engel, Michael S.;Hellenthal, Barbara;Ratcliffe, Brett C.;O'Shea, Brian J.;Schmitt, C. Jonathan;Boufford, David;Morris, Paul J.;Shakya, Subir;Swift, Camm C.;McLeod, Samuel A.;Cohen, Chris M.;Borkent, Christopher J.;Borkent, Christopher J.;Rogers, D. Christopher;Gonzalez, Victor H.;Koenemann, Daniel;Campbell, David;Krause, David W.;Cushing, Paula;Sikes, Derek;Les, Don;DeKeyser, Edward S.;Cruz-Rivera, Edwin;Carter, Emily;Seeholzer, Glenn Fairbanks;Pratt, H. Douglas;Gerwin, John A.;Kaiser, Hinrich;Machio, Irene Villa;Harber, Julian;Zander, Richard H.;Gereau, Roy E.;Applequist, Wendy;Egger, J. Mark;Sagebiel, James Christopher;Lopez-Ribot, Jose L.;Atkinson, Thomas H.;Liebherr, James;Dombroskie, Jason J.;Hoghe, James N.;Wolfe, Jared;McGuire, Jimmy A.;Will, Kipling;Zuparko, Robert L.;Hansen, Robert W.;Brett, Roberta;Leal, Jose H.;Winston, Judith E.;Wen, Jun;Pinheiro, Paulo Duraes Pereira;Lee, Justin L.;Rabeler, Richard;Pargeter, Justin;Gregg, Katharine;Carpenter, Kenneth;McCracken, Kevin;Williams, Kevin;Korneyev, Severyn;Winker, Kevin;Austin, Kyhl A.;Watling, Les;Watson, Linda;Marin, Manuel;Epstein, Marc Edward;Key, Marcus M., Jr.;Marques, Mariana P.;Pyron, Mark;Dovciak, Martin;Hauser, Martin;Sage, Obediah;Garrison, Rosser W.;Winterton, Shaun L.;Gaimari, Stephen D.;West-Eberhard, Mary Jane;Diazgranados, Mauricio;Warren, Mel;Oliver, Michael K.;Ferro, Michael L.;Williams, Kevin;Evenhuis, Neal L.;Snow, Neil;Mizumoto, Nobuaki;Rakes, Patrick;Rohner, Patrick;Rozo-Lopez, Paula;Kerr, Peter H.;Stevens, Peter;Uetz, Peter;Wheeler, Quentin;DeWalt, R. Edward;McClelland, R. Kevan Schoonover;Hellenthal, Ronald;Meier, Rose;Heads, Sam W.;James, Samuel Wooster;Hilty, Steve;Adams, Susan B.;Letcher, Susan G.;Wood, Timothy S.;Walsh, S. J.;Murphy, William L.;Thomas, William Wayt;Khalimov, F. Z.;Rakhimov, Mukhammadtuychi;Abdullajonovich, Shodmonov Feruz;Tashpulatov, Yigitali;Rojas-Runjaic, Fernando J. M.;Cerda Perez, Hugo Gullermo;Torrecilla, Pedro;Zapata, Thirza Ruiz;Hoa, Hoang Quynh;Nguyen, Ngoc;Sharp, Cathy;Fitzpatrick, Moira

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