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Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver

The Active Peptide from Shark Liver (APSL) was expressed in E. coli BL21 cells. The cDNA encoding APSL protein was obtained from shark regenerated hepatic tissue by RT-PCR, then it was cloned in the pET-28a expression vector. The expressed fusion protein was purified by Ni-IDA affinity chromatograph...

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Detalles Bibliográficos
Autores principales: Lv, Zhengbing, Ou, Yu, Li, Qian, Zhang, Wenping, Ye, Boping, Wu, Wutong
Formato: Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2707046/
https://www.ncbi.nlm.nih.gov/pubmed/19597584
http://dx.doi.org/10.3390/md7020258
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author Lv, Zhengbing
Ou, Yu
Li, Qian
Zhang, Wenping
Ye, Boping
Wu, Wutong
author_facet Lv, Zhengbing
Ou, Yu
Li, Qian
Zhang, Wenping
Ye, Boping
Wu, Wutong
author_sort Lv, Zhengbing
collection PubMed
description The Active Peptide from Shark Liver (APSL) was expressed in E. coli BL21 cells. The cDNA encoding APSL protein was obtained from shark regenerated hepatic tissue by RT-PCR, then it was cloned in the pET-28a expression vector. The expressed fusion protein was purified by Ni-IDA affinity chromatography. SDS-PAGE and HPLC analysis showed the purity of the purified fusion protein was more than 98%. The recombinant APSL (rAPSL) was tested for its biological activity both in vitro, by its ability to improve the proliferation of SMMC7721 cells, and in vivo, by its significant protective effects against acute hepatic injury induced by CCl(4) and AAP (acetaminophen) in mice. In addition, the rAPSL could decrease the blood glucose concentration of mice with diabetes mellitus induced by alloxan. Paraffin sections of mouse pancreas tissues showed that rAPSL (3 mg/kg) could effectively protect mouse islets from lesions induced by alloxan, which indicated its potential application in theoretical research and industry.
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spelling pubmed-27070462009-07-08 Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver Lv, Zhengbing Ou, Yu Li, Qian Zhang, Wenping Ye, Boping Wu, Wutong Mar Drugs Article The Active Peptide from Shark Liver (APSL) was expressed in E. coli BL21 cells. The cDNA encoding APSL protein was obtained from shark regenerated hepatic tissue by RT-PCR, then it was cloned in the pET-28a expression vector. The expressed fusion protein was purified by Ni-IDA affinity chromatography. SDS-PAGE and HPLC analysis showed the purity of the purified fusion protein was more than 98%. The recombinant APSL (rAPSL) was tested for its biological activity both in vitro, by its ability to improve the proliferation of SMMC7721 cells, and in vivo, by its significant protective effects against acute hepatic injury induced by CCl(4) and AAP (acetaminophen) in mice. In addition, the rAPSL could decrease the blood glucose concentration of mice with diabetes mellitus induced by alloxan. Paraffin sections of mouse pancreas tissues showed that rAPSL (3 mg/kg) could effectively protect mouse islets from lesions induced by alloxan, which indicated its potential application in theoretical research and industry. Molecular Diversity Preservation International 2009-06-22 /pmc/articles/PMC2707046/ /pubmed/19597584 http://dx.doi.org/10.3390/md7020258 Text en © 2009 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Lv, Zhengbing
Ou, Yu
Li, Qian
Zhang, Wenping
Ye, Boping
Wu, Wutong
Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver
title Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver
title_full Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver
title_fullStr Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver
title_full_unstemmed Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver
title_short Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver
title_sort expression, purification and bioactivities analysis of recombinant active peptide from shark liver
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2707046/
https://www.ncbi.nlm.nih.gov/pubmed/19597584
http://dx.doi.org/10.3390/md7020258
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