title: Next generation synthetic vectors for transformation of the plastid genome of higher plants.
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.
authors.affiliation: Waksman Institute, Rutgers, The State University of New Jersey, 190 Frelinghuysen Road, Piscataway, NJ 08854-8020, USA.
authors.forename: Sugey Ramona
authors.initials: SR
authors.lastname: Sinagawa-García
authors.forename: Tarinee
authors.initials: T
authors.lastname: Tungsuchat-Huang
authors.forename: Octavio
authors.initials: O
authors.lastname: Paredes-López
authors.forename: Pal
authors.initials: P
authors.lastname: Maliga
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000235:genetics / Q000378:metabolism; D015152:Blotting, Northern; D021901:DNA, Intergenic / Q000235:genetics; D004351:Drug Resistance; D004591:Electrophoresis, Polyacrylamide Gel; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics*; D054627:Genome, Plastid / Q000235:genetics*; D015151:Immunoblotting; D010944:Plants / Q000235:genetics*; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics / Q000378:metabolism; D000198:Spectinomycin / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D021901:DNA, Intergenic; D012333:RNA, Messenger; D000198:Spectinomycin; C101625:RbcL protein, plastid; D012273:Ribulose-Bisphosphate Carboxylase; D000103:Acetyl-CoA Carboxylase