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Obpc symposium: maize 2004 & beyond –– recent advances in chloroplast genetic engineering Plant

In vitro cellular & developmental biology 2005 v.41 no.4 pp. 388-404
DNA, Daucus carota, Glycine max, Gossypium hirsutum, Zea mays, agronomic traits, amino acids, antigens, biocompatible materials, biogenesis, biopharmaceuticals, carrots, chloroplast genome, chloroplasts, corn, cotton, crops, disulfide bonds, engineering, enzymes, gene expression, gene silencing, genetic engineering, genetic markers, genetic transformation, genetically modified organisms, in vitro studies, inheritance (genetics), leaves, molecular biology, pleiotropy, proteins, sequence analysis, somatic embryogenesis, soybeans, tobacco, transgenes, vaccines
The chloroplast genetic engineering approach offers a number of unique advantages, including high-level transgene expression, multi-gene engineering in a single transformation event, transgene containment via maternal inheritance, lack of gene silencing, position and pleiotropic effects and undesirable foreign DNA. Thus far, more than 40 transgenes have been stably integrated and expressed via the tobacco chloroplast genome to confer several agronomic traits and produce vaccine antigens, industrially valuable enzymes, biomaterials, and amino acids. Functionality of chloroplast-derived vaccine antigens and therapeutic proteins have been demonstrated by in vitro assays and animal studies. Oral delivery of vaccine antigens has been facilitated by hyperexpression in transgenic chloroplasts (leaves) or non-green plastids (carrots) and the availability of antibiotic-free selectable markers or the ability to excise selectable marker genes. Additionally, the presence of chaperones and enzymes within the chloroplast help to assemble complex multi-subunit proteins and correctly fold proteins containing disulfide bonds, thereby drastically reducing the costs of in vitro processing. Despite such significant progress in chloroplast transformation, this technology has not been extended to major crops. This obstacle emphasizes the need for plastid genome sequencing to increase the efficiency of transformation and conduct basic research in plastid biogenesis and function. However, highly efficient soybean, carrot, and cotton plastid transformation has been recently accomplished via somatic embryogenesis using species-specific chloroplast vectors. Recent advancements facilitate our understanding of plastid biochemistry and molecular biology. This review focuses on exciting recent developments in this field and offers directions for further research and development.