Cargando…

Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis

We set out to assess the NIH/3T3 cell proliferation activity of Arabidopsis oil body-expressed recombinant oleosin–hEGF–hEGF protein. Normally, human epidermal growth factor (hEGF) is purified through complex process, however, oleosin fusion technology provides an inexpensive and scalable platform f...

Descripción completa

Detalles Bibliográficos
Autores principales: Qiang, Weidong, Gao, Tingting, Lan, Xinxin, Guo, Jinnan, Noman, Muhammad, Li, Yaying, Guo, Yongxin, Kong, Jie, Li, Haiyan, Du, Linna, Yang, Jing
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7564230/
https://www.ncbi.nlm.nih.gov/pubmed/32825160
http://dx.doi.org/10.3390/genes11090959
_version_ 1783595666251972608
author Qiang, Weidong
Gao, Tingting
Lan, Xinxin
Guo, Jinnan
Noman, Muhammad
Li, Yaying
Guo, Yongxin
Kong, Jie
Li, Haiyan
Du, Linna
Yang, Jing
author_facet Qiang, Weidong
Gao, Tingting
Lan, Xinxin
Guo, Jinnan
Noman, Muhammad
Li, Yaying
Guo, Yongxin
Kong, Jie
Li, Haiyan
Du, Linna
Yang, Jing
author_sort Qiang, Weidong
collection PubMed
description We set out to assess the NIH/3T3 cell proliferation activity of Arabidopsis oil body-expressed recombinant oleosin–hEGF–hEGF protein. Normally, human epidermal growth factor (hEGF) is purified through complex process, however, oleosin fusion technology provides an inexpensive and scalable platform for its purification. Under a phaseolin promoter, we concatenated oleosin gene to double hEGF (hEGF–hEGF) with plant-preferred codons in the expression vectors and the construct was transformed into Arabidopsis thaliana (Arabidopsis). The transgenic Arabidopsis was validated by RT–PCR and the content of recombinant protein oleosin–hEGF–hEGF was quantified by western blot. Subsequently, the proliferation assay and transdermal absorption were determined by MTT method and immunohistochemical staining, respectively. First, the expression level of hEGF was recorded to be 14.83-ng/μL oil body and due to smaller size transgenic oil bodies expressing the recombinant oleosin–hEGF–hEGF, they were more skin permeable than those of control. Second, via the staining intensity of transgenic oil bodies was greater than EGF at all time points via immunohistochemical staining in transdermal absorption process. Lastly, activity assays of oil bodies expressed oleosin–hEGF–hEGF indicated that they stimulated the NIH/3T3 cell proliferation activity. Our results revealed oil-body-expressed oleosin–hEGF–hEGF was potential new material having implications in the field of medicine.
format Online
Article
Text
id pubmed-7564230
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-75642302020-10-26 Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis Qiang, Weidong Gao, Tingting Lan, Xinxin Guo, Jinnan Noman, Muhammad Li, Yaying Guo, Yongxin Kong, Jie Li, Haiyan Du, Linna Yang, Jing Genes (Basel) Article We set out to assess the NIH/3T3 cell proliferation activity of Arabidopsis oil body-expressed recombinant oleosin–hEGF–hEGF protein. Normally, human epidermal growth factor (hEGF) is purified through complex process, however, oleosin fusion technology provides an inexpensive and scalable platform for its purification. Under a phaseolin promoter, we concatenated oleosin gene to double hEGF (hEGF–hEGF) with plant-preferred codons in the expression vectors and the construct was transformed into Arabidopsis thaliana (Arabidopsis). The transgenic Arabidopsis was validated by RT–PCR and the content of recombinant protein oleosin–hEGF–hEGF was quantified by western blot. Subsequently, the proliferation assay and transdermal absorption were determined by MTT method and immunohistochemical staining, respectively. First, the expression level of hEGF was recorded to be 14.83-ng/μL oil body and due to smaller size transgenic oil bodies expressing the recombinant oleosin–hEGF–hEGF, they were more skin permeable than those of control. Second, via the staining intensity of transgenic oil bodies was greater than EGF at all time points via immunohistochemical staining in transdermal absorption process. Lastly, activity assays of oil bodies expressed oleosin–hEGF–hEGF indicated that they stimulated the NIH/3T3 cell proliferation activity. Our results revealed oil-body-expressed oleosin–hEGF–hEGF was potential new material having implications in the field of medicine. MDPI 2020-08-19 /pmc/articles/PMC7564230/ /pubmed/32825160 http://dx.doi.org/10.3390/genes11090959 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Qiang, Weidong
Gao, Tingting
Lan, Xinxin
Guo, Jinnan
Noman, Muhammad
Li, Yaying
Guo, Yongxin
Kong, Jie
Li, Haiyan
Du, Linna
Yang, Jing
Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis
title Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis
title_full Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis
title_fullStr Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis
title_full_unstemmed Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis
title_short Molecular Pharming of the Recombinant Protein hEGF-hEGF Concatenated with Oleosin Using Transgenic Arabidopsis
title_sort molecular pharming of the recombinant protein hegf-hegf concatenated with oleosin using transgenic arabidopsis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7564230/
https://www.ncbi.nlm.nih.gov/pubmed/32825160
http://dx.doi.org/10.3390/genes11090959
work_keys_str_mv AT qiangweidong molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT gaotingting molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT lanxinxin molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT guojinnan molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT nomanmuhammad molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT liyaying molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT guoyongxin molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT kongjie molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT lihaiyan molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT dulinna molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis
AT yangjing molecularpharmingoftherecombinantproteinhegfhegfconcatenatedwitholeosinusingtransgenicarabidopsis