Wang Yun-Peng, Wei Zheng-Yi, Zhong Xiao-Fang, Lin Chun-Jing, Cai Yu-Hong, Ma Jian, Zhang Yu-Ying, Liu Yan-Zhi, Xing Shao-Chen
Jilin Provincial Key Laboratory of Agricultural Biotechnology, Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, No. 1363, Shengtai st., Changchun 130033, China.
Institute of Agricultural Quality Standard and Testing Technology, Jilin Academy of Agricultural Sciences, No. 1363, Shengtai st., Changchun 130033, China.
Int J Mol Sci. 2015 Dec 23;17(1):19. doi: 10.3390/ijms17010019.
Basic fibroblast growth factor (bFGF) is a multifunctional factor in acceleration of cell proliferation, differentiation and transference, and therefore widely used in clinical applications. In this study, expression vector pWX-Nt03 harboring a codon-optimized bFGF gene was constructed and introduced into the tobacco chloroplasts by particle bombardment. After four rounds of selection, bFGF was proved to integrate into the chloroplast genome of regenerated plants and two of four transgenic plants were confirmed to be homoplastomic by PCR and Southern hybridization. ELISA assay indicated that bFGF represented approximately 0.1% of total soluble protein in the leaves of transplastomic tobacco plants. This is the first report of bFGF expression via chloroplast transformation in model plant, providing an additional option for the production of chloroplast-produced therapeutic proteins.
碱性成纤维细胞生长因子（bFGF）是一种促进细胞增殖、分化和转移的多功能因子，因此在临床应用中被广泛使用。在本研究中，构建了携带密码子优化的bFGF基因的表达载体pWX-Nt03，并通过粒子轰击将其导入烟草叶绿体。经过四轮筛选，证明bFGF整合到再生植物的叶绿体基因组中，通过PCR和Southern杂交确认四株转基因植物中有两株为同质性叶绿体。ELISA分析表明，bFGF在转基因烟草植物叶片的总可溶性蛋白中约占0.1%。这是在模式植物中通过叶绿体转化表达bFGF的首次报道，为叶绿体生产治疗性蛋白提供了另一种选择。
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