Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants

Guo Guang Zheng; Ying Hua Yang; Qing Rao; Yong Min Lin; Bin Zhang; Ke Fu Wu

doi:10.1016/j.pep.2005.07.018

Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants

Guo Guang Zheng^*, Ying Hua Yang, Qing Rao, Yong Min Lin, Bin Zhang, Ke Fu Wu

^*Corresponding author for this work

Medicine, Hematology Oncology Division

Research output: Contribution to journal › Article › peer-review

4 Scopus citations

Abstract

The cDNA encoding N-terminal three immunoglobin-like domains of human M-CSFR was linked to His-tag and endoplasmic reticulum retention sequence (KDEL) before being inserted into the genome of tobacco plant, Nicotiana tabacum cv. NC-89, by Agrobacterium tumefaciens-mediated transformation. The insertion and expression of target gene were confirmed by PCR, ELISA, and Western blot. The recombinant M-CSFsR reached a maximum expression level of 1.92% of total soluble protein in transgenic tobacco plant leaf tissues. The recombinant M-CSFsR could be purified through a one-step IMAC process and its bioactivity was confirmed by the inhibition of colony formation of J6-1 cells. The results suggested that we successfully expressed a high level of bioactive human M-CSFsR in tobacco plants.

Original language	English (US)
Pages (from-to)	367-373
Number of pages	7
Journal	Protein Expression and Purification
Volume	46
Issue number	2
DOIs	https://doi.org/10.1016/j.pep.2005.07.018
State	Published - Apr 2006

Keywords

Bioactive
Macrophage colony-stimulating factor
Soluble receptor
Tobacco
Transgenic plants

ASJC Scopus subject areas

Biotechnology

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10.1016/j.pep.2005.07.018

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@article{ec0351980dda4dd7b85047a9bacdaa29,

title = "Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants",

abstract = "The cDNA encoding N-terminal three immunoglobin-like domains of human M-CSFR was linked to His-tag and endoplasmic reticulum retention sequence (KDEL) before being inserted into the genome of tobacco plant, Nicotiana tabacum cv. NC-89, by Agrobacterium tumefaciens-mediated transformation. The insertion and expression of target gene were confirmed by PCR, ELISA, and Western blot. The recombinant M-CSFsR reached a maximum expression level of 1.92% of total soluble protein in transgenic tobacco plant leaf tissues. The recombinant M-CSFsR could be purified through a one-step IMAC process and its bioactivity was confirmed by the inhibition of colony formation of J6-1 cells. The results suggested that we successfully expressed a high level of bioactive human M-CSFsR in tobacco plants.",

keywords = "Bioactive, Macrophage colony-stimulating factor, Soluble receptor, Tobacco, Transgenic plants",

author = "Zheng, {Guo Guang} and Yang, {Ying Hua} and Qing Rao and Lin, {Yong Min} and Bin Zhang and Wu, {Ke Fu}",

note = "Funding Information: This work was supported by the Tianjin Science and Technology Development program of China (003119311) and National Natural Science Foundation of China (30470897). We thank Dr. Ge Li, Zhen-Yu Cao, and Xiu-Jun Zhang for their technical help.",

year = "2006",

month = apr,

doi = "10.1016/j.pep.2005.07.018",

language = "English (US)",

volume = "46",

pages = "367--373",

journal = "Protein Expression and Purification",

issn = "1046-5928",

publisher = "Academic Press Inc.",

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}

TY - JOUR

T1 - Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants

AU - Zheng, Guo Guang

AU - Yang, Ying Hua

AU - Rao, Qing

AU - Lin, Yong Min

AU - Zhang, Bin

AU - Wu, Ke Fu

N1 - Funding Information: This work was supported by the Tianjin Science and Technology Development program of China (003119311) and National Natural Science Foundation of China (30470897). We thank Dr. Ge Li, Zhen-Yu Cao, and Xiu-Jun Zhang for their technical help.

PY - 2006/4

Y1 - 2006/4

N2 - The cDNA encoding N-terminal three immunoglobin-like domains of human M-CSFR was linked to His-tag and endoplasmic reticulum retention sequence (KDEL) before being inserted into the genome of tobacco plant, Nicotiana tabacum cv. NC-89, by Agrobacterium tumefaciens-mediated transformation. The insertion and expression of target gene were confirmed by PCR, ELISA, and Western blot. The recombinant M-CSFsR reached a maximum expression level of 1.92% of total soluble protein in transgenic tobacco plant leaf tissues. The recombinant M-CSFsR could be purified through a one-step IMAC process and its bioactivity was confirmed by the inhibition of colony formation of J6-1 cells. The results suggested that we successfully expressed a high level of bioactive human M-CSFsR in tobacco plants.

AB - The cDNA encoding N-terminal three immunoglobin-like domains of human M-CSFR was linked to His-tag and endoplasmic reticulum retention sequence (KDEL) before being inserted into the genome of tobacco plant, Nicotiana tabacum cv. NC-89, by Agrobacterium tumefaciens-mediated transformation. The insertion and expression of target gene were confirmed by PCR, ELISA, and Western blot. The recombinant M-CSFsR reached a maximum expression level of 1.92% of total soluble protein in transgenic tobacco plant leaf tissues. The recombinant M-CSFsR could be purified through a one-step IMAC process and its bioactivity was confirmed by the inhibition of colony formation of J6-1 cells. The results suggested that we successfully expressed a high level of bioactive human M-CSFsR in tobacco plants.

KW - Bioactive

KW - Macrophage colony-stimulating factor

KW - Soluble receptor

KW - Tobacco

KW - Transgenic plants

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SN - 1046-5928

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EP - 373

JO - Protein Expression and Purification

JF - Protein Expression and Purification

IS - 2

ER -

Expression of bioactive human M-CSF soluble receptor in transgenic tobacco plants

Abstract

Keywords

ASJC Scopus subject areas

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