Kumar Shashi, Daniell Henry
Department of Molecular Biology and Microbiology, University of Central Florida, Orlando, USA.
Methods Mol Biol. 2004;267:365-83. doi: 10.1385/1-59259-774-2:365.
The chloroplast genome is ideal for engineering because it offers a number of attractive advantages, including high-level gene expression, the feasibility of expressing multiple genes or pathways in a single transformation event, and transgene containment due to lack of pollen transmission. The chloroplast-based expression system is suitable for hyperexpression of foreign proteins, oral delivery of vaccine antigens and therapeutic proteins, via both leaves and fruits. Through the refinement of expression vectors and use of chaperones, chloroplasts produce up to 47% of foreign protein in the total cellular protein in transgenic tissues. This chapter describes various techniques for creating chloroplast transgenic plants and their biochemical and molecular characterization. Suitable examples for application of chloroplast genetic engineering in human medicine are provided.
叶绿体基因组对于基因工程而言是理想的,因为它具有许多吸引人的优势,包括高水平的基因表达、在单次转化事件中表达多个基因或途径的可行性,以及由于缺乏花粉传播而实现的转基因隔离。基于叶绿体的表达系统适用于外源蛋白的超量表达、通过叶片和果实进行疫苗抗原和治疗性蛋白的口服递送。通过优化表达载体和使用伴侣蛋白,叶绿体在转基因组织的总细胞蛋白中产生高达47%的外源蛋白。本章描述了创建叶绿体转基因植物的各种技术及其生化和分子特征。还提供了叶绿体基因工程在人类医学中应用的合适示例。