Lecture 10 - PLB 113 Lecture 10 III Gene Transfer and...

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PLB 113 – Lecture 10 III. Gene Transfer and Epigenetics E. Cosuppression IV. Chloroplast Biogenesis A. Chloroplast function B. Developmental interconversions C. Chloroplast structure D. Semiautonomous organelle T-DNA transformation isn’t the only way of getting genes into the plant. Microinjection of gene can be applied as well. - take a gene and put into a plant cell - that plant that contains the plant cell is hemizygous for the transgene because the corresponding locus isn’t in the plant chromosome. (since transgene is the new gene in the plant) - self-fertilize the hemizygous plant to produce homozygous plant ex. L- x L- = LL, L-, and - - Clicker: What percentage of the progeny from this transgeneic plant should be Kanamysin- resistant? Ans. 75% In this case, L represents the dominant Lectin gene, it also represents T-DNA. In addition, T-DNA also contains the selectable marker (Kan.- resistant), NPTII. So, NPTII acts as dominant trait. Whenever you have the T-DNA, you have the NPTII. So, a single locus (single insertion to the plant) which the T-DNA is inserted, then you will have 75% of the progeny that are homozygous. ----- T-DNA contains selectable marker and OCS (octopine synthase gene) - use this T-DNA to transform numbers of different plants - each of the transgenic plants will express OCS at different quantitative levels - rationalize: the level of expression of transgene depends on where the transgene is integrated into the genome. Question of the day: What’s the phenotype(s) of the plants infected with an Agrobacterium containing the following T-DNA? Everything between LB and RB will be transferred. So, there are two options here: 1. transfer from all the way across => tumor with root that’s producing nopaline a. This DNA only has TMS gene but not TMR gene, so it’s treated as TMS with tmr mutant.
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