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A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3)

An apoptosis correlated molecule—protein Latcripin-1 of Lentinula edodes C(91–3)—was expressed and characterized in Pichia pastoris GS115. The total RNA was obtained from Lentinula edodes C(91–3). According to the transcriptome, the full-length gene of Latcripin-1 was isolated with 3′-Full Rapid Amp...

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Detalles Bibliográficos
Autores principales: Liu, Ben, Zhong, Mintao, Lun, Yongzhi, Wang, Xiaoli, Sun, Wenchang, Li, Xingyun, Ning, Anhong, Cao, Jing, Zhang, Wei, Liu, Lei, Huang, Min
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3382792/
https://www.ncbi.nlm.nih.gov/pubmed/22754362
http://dx.doi.org/10.3390/ijms13056246
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author Liu, Ben
Zhong, Mintao
Lun, Yongzhi
Wang, Xiaoli
Sun, Wenchang
Li, Xingyun
Ning, Anhong
Cao, Jing
Zhang, Wei
Liu, Lei
Huang, Min
author_facet Liu, Ben
Zhong, Mintao
Lun, Yongzhi
Wang, Xiaoli
Sun, Wenchang
Li, Xingyun
Ning, Anhong
Cao, Jing
Zhang, Wei
Liu, Lei
Huang, Min
author_sort Liu, Ben
collection PubMed
description An apoptosis correlated molecule—protein Latcripin-1 of Lentinula edodes C(91–3)—was expressed and characterized in Pichia pastoris GS115. The total RNA was obtained from Lentinula edodes C(91–3). According to the transcriptome, the full-length gene of Latcripin-1 was isolated with 3′-Full Rapid Amplification of cDNA Ends (RACE) and 5′-Full RACE methods. The full-length gene was inserted into the secretory expression vector pPIC9K. The protein Latcripin-1 was expressed in Pichia pastoris GS115 and analyzed by Sodium Dodecylsulfonate Polyacrylate Gel Electrophoresis (SDS-PAGE) and Western blot. The Western blot showed that the protein was expressed successfully. The biological function of protein Latcripin-1 on A549 cells was studied with flow cytometry and the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyl-tetrazolium Bromide (MTT) method. The toxic effect of protein Latcripin-1 was detected with the MTT method by co-culturing the characterized protein with chick embryo fibroblasts. The MTT assay results showed that there was a great difference between protein Latcripin-1 groups and the control group (p < 0.05). There was no toxic effect of the characterized protein on chick embryo fibroblasts. The flow cytometry showed that there was a significant difference between the protein groups of interest and the control group according to apoptosis function (p < 0.05). At the same time, cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. The work demonstrates that protein Latcripin-1 can induce apoptosis of human lung cancer cells A549 and brings new insights into and advantages to finding anti-tumor proteins.
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spelling pubmed-33827922012-06-29 A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3) Liu, Ben Zhong, Mintao Lun, Yongzhi Wang, Xiaoli Sun, Wenchang Li, Xingyun Ning, Anhong Cao, Jing Zhang, Wei Liu, Lei Huang, Min Int J Mol Sci Article An apoptosis correlated molecule—protein Latcripin-1 of Lentinula edodes C(91–3)—was expressed and characterized in Pichia pastoris GS115. The total RNA was obtained from Lentinula edodes C(91–3). According to the transcriptome, the full-length gene of Latcripin-1 was isolated with 3′-Full Rapid Amplification of cDNA Ends (RACE) and 5′-Full RACE methods. The full-length gene was inserted into the secretory expression vector pPIC9K. The protein Latcripin-1 was expressed in Pichia pastoris GS115 and analyzed by Sodium Dodecylsulfonate Polyacrylate Gel Electrophoresis (SDS-PAGE) and Western blot. The Western blot showed that the protein was expressed successfully. The biological function of protein Latcripin-1 on A549 cells was studied with flow cytometry and the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyl-tetrazolium Bromide (MTT) method. The toxic effect of protein Latcripin-1 was detected with the MTT method by co-culturing the characterized protein with chick embryo fibroblasts. The MTT assay results showed that there was a great difference between protein Latcripin-1 groups and the control group (p < 0.05). There was no toxic effect of the characterized protein on chick embryo fibroblasts. The flow cytometry showed that there was a significant difference between the protein groups of interest and the control group according to apoptosis function (p < 0.05). At the same time, cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. The work demonstrates that protein Latcripin-1 can induce apoptosis of human lung cancer cells A549 and brings new insights into and advantages to finding anti-tumor proteins. Molecular Diversity Preservation International (MDPI) 2012-05-21 /pmc/articles/PMC3382792/ /pubmed/22754362 http://dx.doi.org/10.3390/ijms13056246 Text en © 2012 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
Liu, Ben
Zhong, Mintao
Lun, Yongzhi
Wang, Xiaoli
Sun, Wenchang
Li, Xingyun
Ning, Anhong
Cao, Jing
Zhang, Wei
Liu, Lei
Huang, Min
A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3)
title A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3)
title_full A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3)
title_fullStr A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3)
title_full_unstemmed A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3)
title_short A Novel Apoptosis Correlated Molecule: Expression and Characterization of Protein Latcripin-1 from Lentinula edodes C(91–3)
title_sort novel apoptosis correlated molecule: expression and characterization of protein latcripin-1 from lentinula edodes c(91–3)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3382792/
https://www.ncbi.nlm.nih.gov/pubmed/22754362
http://dx.doi.org/10.3390/ijms13056246
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