Chloroplasts are a useful platform for the expression of recombinant proteins in higher plants. Transgenes can be introduced into the plastid genome (plastome) either by PEG transformation of plant protoplasts, or, more commonly, by the biolistic method, using leaves or suspension cells. Transgenes are integrated by double recombination events between flanking sequences in the vector and homologous sequences in the plastome. The genetic engineering of the plastome allows high-level foreign protein expression, site-specific gene integration, expression of multiple genes as operons, marker gene excision, and transgene containment. Since the first example of stable plastid transformation in higher plants, methods for DNA introduction, marker genes and selection strategies, vector types, and methods for marker excision have been improved. Although the plastids of some species remain difficult to transform, positive results have been shown for about 20 species. In this chapter, we summarize the basic structural and expression features of the plastid genome of higher plants, and discuss the development of a number of innovative enabling technologies for plastome transformation, the most recent and significant biotechnological applications, and the future perspectives of this technology.
The chloroplasts as platform for recombinant proteins production
Scotti N;Bellucci M;Cardi T
2013
Abstract
Chloroplasts are a useful platform for the expression of recombinant proteins in higher plants. Transgenes can be introduced into the plastid genome (plastome) either by PEG transformation of plant protoplasts, or, more commonly, by the biolistic method, using leaves or suspension cells. Transgenes are integrated by double recombination events between flanking sequences in the vector and homologous sequences in the plastome. The genetic engineering of the plastome allows high-level foreign protein expression, site-specific gene integration, expression of multiple genes as operons, marker gene excision, and transgene containment. Since the first example of stable plastid transformation in higher plants, methods for DNA introduction, marker genes and selection strategies, vector types, and methods for marker excision have been improved. Although the plastids of some species remain difficult to transform, positive results have been shown for about 20 species. In this chapter, we summarize the basic structural and expression features of the plastid genome of higher plants, and discuss the development of a number of innovative enabling technologies for plastome transformation, the most recent and significant biotechnological applications, and the future perspectives of this technology.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
