Tissue Cultures Suspension cultures of the wild-type and the transplastomic plants were initiated by germinating seeds in solid Murashige and Skoog medium containing 1. These results demonstrate that the ASA2 gene is incorporated into the plastid genome in our transformed plants and that the insertions are identical to each other in the four pAST-I lines and in the three pAST-IV lines. Thus far, a few foreign genes have been expressed that provide, for example, resistance to insects McBride et al. The following symbols are legal: All transplastomic seedlings were green and control progeny were white. The resistance to the selection agent spectinomycin and to 5-methyl-Trp was transmitted maternally.
The variation in the Trp level between different transplastomic plants Fig. Here we report the creation and analysis of ASA2 transplastomic plants for which the ASA2 transgene was independently targeted into two separate regions of the plastid genome. Transient foreign gene expression in chloroplasts of cultured tobacco cells after biolistic delivery of chloroplast vectors. Probes used for RNA-blot hybridizations are indicated. Effect of 5MT on the germination and growth of seedlings. Northern-blot hybridizations were carried out to determine the expression of the Prrn- ASA2 gene in the transformed plants.
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A revised medium for rapid growth and bioassay with tobacco cultures. Increasing tryptophan synthesis in a forage legume Astragalus sinicus by expressing the tobacco feedback-insensitive anthranilate synthase ASA2 gene. This indicates that incorporation of the aadA gene alone in the plastid genome did not affect the leaf Trp level. Big booty mom leaves the step son to fuck her hard 7: It is clear that in the case of tobacco, there would probably not be a problem with subunit interaction since the ASA2 gene is from the same plant. It is known that the nuclear and plastid genomes have a unique base composition and codon usage Oliver et al. Our studies demonstrate, to our knowledge, the first example of introducing a native nuclear gene of presumed pre-endosymbiotic origin into plastids to modify an endogenous biosynthetic pathway.
Therefore, the naturally occurring ASA2 gene appears to be expressed at a very low level or in a small population of leaf cells in the wild-type plants, reflecting tight regulation at the transcription level for this isoform of the first enzyme in the Trp biosynthetic pathway. We also tested the 5MT tolerance of suspension cultures initiated from the wild-type and transplastomic plants. Xing-Hai Zhang , Jeffrey E. AS protein analysis of transplastomic plants. Transformation of tobacco leaves was carried out as described by Svab and Maliga using 0. A similar modest increase in K i was observed by Cho et al. Young, expanding leaves from the same location on different plants of the same age and condition and ones grown in the same growth chamber were analyzed for free Trp content.