Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene
Main Author: | |
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Publication Date: | 2010 |
Other Authors: | , , , , , |
Format: | Article |
Language: | eng |
Source: | Brazilian Journal of Medical and Biological Research |
Download full: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000100007 |
Summary: | Myocardial ischemic preconditioning up-regulated protein 1 (Mipu1), a novel zinc finger protein, was originally cloned using bioinformatic analysis and 5' RACE technology of rat heart after a transient myocardial ischemia/reperfusion procedure in our laboratory. In order to investigate the functions of Mipu1, the recombinant prokaryotic expression vector pQE31-Mipu1 was constructed and transformed into Escherichia coli M15(pREP4), and Mipu1-6His fusion protein was expressed and purified. The identity of the purified protein was confirmed by mass spectrometry. The molecular mass of the Mipu1 protein was 70.03779 kDa. The fusion protein was intracutaneously injected to immunize New Zealand rabbits to produce a polyclonal antibody. The antibody titer was approximately 1:16,000. The antibody was tested by Western blotting for specificity and sensitivity. Using the antibody, it was found that Mipu1 was highly expressed in the heart and brain of rats and was localized in the nucleus of H9c2 myogenic cells. The present study lays the foundation for further study of the biological functions of Mipu1. |
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Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related geneMipu1PurificationExpressionPolyclonal antibodyMyocardial ischemic preconditioning up-regulated protein 1 (Mipu1), a novel zinc finger protein, was originally cloned using bioinformatic analysis and 5' RACE technology of rat heart after a transient myocardial ischemia/reperfusion procedure in our laboratory. In order to investigate the functions of Mipu1, the recombinant prokaryotic expression vector pQE31-Mipu1 was constructed and transformed into Escherichia coli M15(pREP4), and Mipu1-6His fusion protein was expressed and purified. The identity of the purified protein was confirmed by mass spectrometry. The molecular mass of the Mipu1 protein was 70.03779 kDa. The fusion protein was intracutaneously injected to immunize New Zealand rabbits to produce a polyclonal antibody. The antibody titer was approximately 1:16,000. The antibody was tested by Western blotting for specificity and sensitivity. Using the antibody, it was found that Mipu1 was highly expressed in the heart and brain of rats and was localized in the nucleus of H9c2 myogenic cells. The present study lays the foundation for further study of the biological functions of Mipu1.Associação Brasileira de Divulgação Científica2010-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000100007Brazilian Journal of Medical and Biological Research v.43 n.1 2010reponame:Brazilian Journal of Medical and Biological Researchinstname:Associação Brasileira de Divulgação Científica (ABDC)instacron:ABDC10.1590/S0100-879X2009005000010info:eu-repo/semantics/openAccessWang,G.Zuo,X.Jiang,L.Wang,K.Wei,X.Zhang,B.Xiao,X.eng2010-01-13T00:00:00Zoai:scielo:S0100-879X2010000100007Revistahttps://www.bjournal.org/https://old.scielo.br/oai/scielo-oai.phpbjournal@terra.com.br||bjournal@terra.com.br1414-431X0100-879Xopendoar:2010-01-13T00:00Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC)false |
dc.title.none.fl_str_mv |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene |
title |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene |
spellingShingle |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene Wang,G. Mipu1 Purification Expression Polyclonal antibody |
title_short |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene |
title_full |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene |
title_fullStr |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene |
title_full_unstemmed |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene |
title_sort |
Tissue expression and subcellular localization of Mipu1, a novel myocardial ischemia-related gene |
author |
Wang,G. |
author_facet |
Wang,G. Zuo,X. Jiang,L. Wang,K. Wei,X. Zhang,B. Xiao,X. |
author_role |
author |
author2 |
Zuo,X. Jiang,L. Wang,K. Wei,X. Zhang,B. Xiao,X. |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Wang,G. Zuo,X. Jiang,L. Wang,K. Wei,X. Zhang,B. Xiao,X. |
dc.subject.por.fl_str_mv |
Mipu1 Purification Expression Polyclonal antibody |
topic |
Mipu1 Purification Expression Polyclonal antibody |
description |
Myocardial ischemic preconditioning up-regulated protein 1 (Mipu1), a novel zinc finger protein, was originally cloned using bioinformatic analysis and 5' RACE technology of rat heart after a transient myocardial ischemia/reperfusion procedure in our laboratory. In order to investigate the functions of Mipu1, the recombinant prokaryotic expression vector pQE31-Mipu1 was constructed and transformed into Escherichia coli M15(pREP4), and Mipu1-6His fusion protein was expressed and purified. The identity of the purified protein was confirmed by mass spectrometry. The molecular mass of the Mipu1 protein was 70.03779 kDa. The fusion protein was intracutaneously injected to immunize New Zealand rabbits to produce a polyclonal antibody. The antibody titer was approximately 1:16,000. The antibody was tested by Western blotting for specificity and sensitivity. Using the antibody, it was found that Mipu1 was highly expressed in the heart and brain of rats and was localized in the nucleus of H9c2 myogenic cells. The present study lays the foundation for further study of the biological functions of Mipu1. |
publishDate |
2010 |
dc.date.none.fl_str_mv |
2010-01-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000100007 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000100007 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0100-879X2009005000010 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica |
publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica |
dc.source.none.fl_str_mv |
Brazilian Journal of Medical and Biological Research v.43 n.1 2010 reponame:Brazilian Journal of Medical and Biological Research instname:Associação Brasileira de Divulgação Científica (ABDC) instacron:ABDC |
instname_str |
Associação Brasileira de Divulgação Científica (ABDC) |
instacron_str |
ABDC |
institution |
ABDC |
reponame_str |
Brazilian Journal of Medical and Biological Research |
collection |
Brazilian Journal of Medical and Biological Research |
repository.name.fl_str_mv |
Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC) |
repository.mail.fl_str_mv |
bjournal@terra.com.br||bjournal@terra.com.br |
_version_ |
1754302938085326848 |