Study on human aFGF fusion gene transformation with soybean 24 kDa oleosin and expression in safflower.
- Author:
Yu-Hong GAI
1
;
Wang WANG
;
Xiao JIN
;
Jia-Qi WANG
;
Hong-Bo LI
;
Gang WANG
;
Jing YANG
;
Jian WEI
Author Information
- Publication Type:Journal Article
- MeSH: Agrobacterium tumefaciens; genetics; Carthamus tinctorius; genetics; Fibroblast Growth Factor 1; genetics; Gene Expression Regulation, Plant; Genetic Vectors; Humans; Membrane Proteins; genetics; Plant Proteins; genetics; Plants, Genetically Modified; Polymerase Chain Reaction; Transformation, Genetic
- From: China Journal of Chinese Materia Medica 2013;38(12):1898-1904
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the expression of acidic fibroblast growth factor (aFGF) in transgenic safflower and lay the foundation for the use of the plant bioreactor large-scale production aFGF.
METHODThe haFGF gene was transformed into plant preference of the aFGF sequence as a basis for design of primers, plant preferences aFGF gene sequences was amplified by PCR. The vegetable body expression vector was constructed by using digested connection method and then transferred to Agrobacterium tumefaciens EHA105 by the freeze-thaw method. It transferred to safflowers by agrobacterium-mediated transformation method, and identified by PCR, southern blot and RT-PCR.
RESULTThe full-length aFGF gene sequences were amplified through PCR and constructed into plant expression vector with soybean oleosin and promoter, and transformed into safflower. Three independently transformed safflower plant units with point insertion were successfully obtained, which showed the same size of aFGF expression at the transcriptional level.
CONCLUSIONThe plant oil body expression vectors were successfully constructed, and the optimal condition for genetic transformation was selected. The transgenic safflower plants were obtained.