Title

A protocol for expression of foreign genes in chloroplasts

Authors

Authors

D. Verma; N. P. Samson; V. Koya;H. Daniell

Comments

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Abbreviated Journal Title

Nat. Protoc.

Keywords

TRANSGENIC TOBACCO CHLOROPLASTS; STABLE PLASTID TRANSFORMATION; PLANT-BASED VACCINE; LACTUCA-SATIVA L.; NICOTIANA-TABACUM; TRANSPLASTOMIC TOBACCO; MATERNAL INHERITANCE; THERAPEUTIC PROTEINS; CONFERS RESISTANCE; PROTECTIVE ANTIGEN; Biochemical Research Methods

Abstract

Several major costs associated with the production of biopharmaceuticals or vaccines in fermentation-based systems could be minimized by using plant chloroplasts as bioreactors, which facilitates rapid scale-up. Oral delivery of chloroplast-derived therapeutic proteins through plant cells eliminates expensive purification steps, low temperature storage, transportation and sterile injections for their delivery. Chloroplast transformation technology (CTT) has also been successfully used to engineer valuable agronomic traits and for the production of industrial enzymes and biomaterials. Here, we provide a detailed protocol for the construction of chloroplast expression and integration vectors, selection and regeneration of transformants, evaluation of transgene integration and inheritance, confirmation of transgene expression and extraction, and quantitation and purification of foreign proteins. Integration of appropriate transgenes into chloroplast genomes and the resulting high levels of functional protein expression can be achieved in similar to 6 months in lettuce and tobacco. CTT is eco-friendly because transgenes are maternally inherited in most crop plants.

Journal Title

Nature Protocols

Volume

3

Issue/Number

4

Publication Date

1-1-2008

Document Type

Article

Language

English

First Page

739

Last Page

758

WOS Identifier

WOS:000254914500020

ISSN

1754-2189

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