编码子优化基因在米细胞悬浮培养中产生活跃的人类基本纤维细胞生长因子
Meysam Bastami1, Ramin Hosseini1
1Department of Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Iran.
Growth factors (Chur, Switzerland)
|November 1, 2024
概括
研究人员优化了人类基本纤维细胞生长因子 (hbFGF) 的米表达. 转基因大米细胞成功地产生和分泌了生物活性hbFGF,证明了其作为重组蛋白质生产系统的潜力.
科学领域:
- 植物生物技术 植物生物技术
- 分子生物学分子生物学
- 生物化学 生化学
背景情况:
- 人类基本纤维细胞生长因子 (hbFGF) 对于细胞生长和组织修复至关重要.
- 开发用于重组蛋白质生产的高效系统对于治疗应用至关重要.
- 基于植物的表达系统为传统方法提供了可扩展和具有成本效益的替代方案.
研究的目的:
- 为了优化hbFGF的编码序列,在大米中增强表达.
- 开发一种稳定的转基因大米细胞系,用于高水平的hbFGF的生产和分泌.
- 为了评估来自大米的生物活性hbFGF.
主要方法:
- 对大米 hbFGF 序列的 Codon 优化.
- 使用RAmy3D促进剂和信号构建表达盒.
- 用Agrobacterium tumefaciens LBA 4404.4 的方法转化大米.
- 通过ELISA选择高表达系并建立细胞悬浮培养.
- 通过Western blot和SDS-PAGE确认hbFGF的表达和分泌.
主要成果:
- 成功生成了对大米表达的编码子优化的hbFGF序列.
- 建立了一个转基因大米细胞系,表现出高水平的hbFGF表达 (38.1 mg/kg新鲜体重).
- 在培养基中显示功能性hbFGF的分泌.
- 在刺激NIH/3T3细胞增殖方面,确认了来自大米的hbFGF与商业hbFGF的可比生物活性.
结论:
- 米可以被设计成一种高效的生物反应器,用于生产重组的人类基本纤维细胞生长因子.
- 开发的转基因大米细胞系为可扩展和具有成本效益的生物活性hbFGF生产提供了一个有前途的平台.
- 这项研究强调了植物分子农业在治疗性蛋白质生成方面的潜力.
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