Recombinant keratinocyte growth factor 1 in tobacco potentially promotes wound healing in diabetic rats.
Keratinocyte growth factor 1 (KGF1) is a growth factor that promotes epidermal cell proliferation, migration, differentiation, and wound repair. It is expressed at low levels in a form of inclusion body in E. coli. In order to increase its expression and activity, we produced tobacco plants expressing KGF1 via Agrobacterium-mediated transformation using a potato virus X (PVX)-based vector (pgR107). The vector contained the sequence encoding the KGF1 gene fused with a green florescence protein. The recombinant plasmid was introduced into leaf cells of Nicotiana benthamiana (a wild Australian tobacco) via Agrobacterium-mediated agroinfiltration. As determined by fluorescence and Western blot of leaf extracts, the KGF1 gene was correctly translated into the tobacco plants. The recombinant KGF1 was purified from plant tissues by heparin affinity chromatography, and cell proliferation in NIH/3T3 cells was stimulated by the purified KGF1. The purified KGF1 was also applied to the wounds of type-II diabetic rats. KGF1 had accumulated to levels as high as 530  μ g/g fresh weight in the leaves of agroinfected plants. We show that plant-derived KGF1 can promote the proliferation of NIH/3T3 cells and have significant effects on the type-II diabetic rat. The present findings indicated that KGF1 from tobacco maintains its biological activity, implying prospective industrial production in a plant bioreactor.
College of Life Science, Anhui Agricultural University, Hefei 230036, China.
Zhi-Guo
ZG
Feng
Department of Biology, Guangdong Medical College, Dongguang 523808, China.
Shi-Feng
SF
Pang
Department of General Surgery, Cixi People's Hospital, Ningbo 315300, China.
Ding-Jiong
DJ
Guo
Traumatic Medicine Center, Lishui People's Hospital, Lishui 315300, China.
Yue-Tao
YT
Yang
Traumatic Medicine Center, Lishui People's Hospital, Lishui 315300, China.
Bin
B
Liu
Traumatic Medicine Center, Lishui People's Hospital, Lishui 315300, China.
Ji-Wei
JW
Wang
Department of Biology, Guangdong Medical College, Dongguang 523808, China.
Ke-Qin
KQ
Zheng
College of Life Science, Anhui Agricultural University, Hefei 230036, China.
Yi
Y
Lin
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects*; D003921:Diabetes Mellitus, Experimental; D005260:Female; D051523:Fibroblast Growth Factor 7 / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D030821:Plants, Genetically Modified / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D014026:Nicotiana / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects*
C496453:FGF7 protein, human; D011993:Recombinant Fusion Proteins; D051523:Fibroblast Growth Factor 7