Next generation synthetic vectors for transformation of the plastid genome of higher plants |
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Authors: | Sugey Ramona Sinagawa-García Tarinee Tungsuchat-Huang Octavio Paredes-López Pal Maliga |
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Institution: | (1) Waksman Institute, Rutgers, The State University of New Jersey, 190 Frelinghuysen, Road, Piscataway, NJ 08854-8020, USA;(2) Depto. de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del IPN, Apdo. Postal 629, 36500 Irapuato, Gto., Mexico |
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Abstract: | Plastid transformation vectors are E. coli plasmids carrying a plastid marker gene for selection, adjacent cloning sites and flanking plastid DNA to target insertions
in the plastid genome by homologous recombination. We report here on a family of next generation plastid vectors carrying
synthetic DNA vector arms targeting insertions in the rbcL-accD intergenic region of the tobacco (Nicotiana tabacum) plastid genome. The pSS22 plasmid carries only synthetic vector arms from which the undesirable restriction sites have been
removed by point mutations. The pSS24 vector carries a c-Myc tagged spectinomycin resistance (aadA) marker gene whereas in vector pSS30 aadA is flanked with loxP sequences for post-transformation marker excision. The synthetic vectors will enable direct manipulation of passenger genes
in the transformation vector targeting insertions in the rbcL-accD intergenic region that contains many commonly used restriction sites.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Homologous recombination Nicotiana tabacum Plastid transformation Tobacco Vector |
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