Project Details
Description
Recombinant DNA and plant tissue culture selection techniques will be used
to develop modified versions of Cauliflower mosaic virus (CaMV)DNA. It is
intended that the final modified version be capable of transferring new DNA
sequences or genes into maize tissue culture cells where stabilization and
expression of such genes could occur. Our approach is to insert a
bacterial drug resistance marker gene into a nonessential region of the
CaMV genome using synthetic primers, in vitro strand synthesis, and M13
cloning techniques. The marker gene will be introduced into plant tissue
culture cells and drug-resistant cells selected. We will further modify
the marker CaMV genome by in vitro mutagenesis and cloning into E. coli
using the plasmid pBR322 to identify other potentially nonessential regions
of the CaMV genome. These regions will be deleted to the extent possible;
their location, size, and the sequence of the new constructions will be
determined by M13 sequencing. The drug resistance marker will be
maintained by selection in E. coli and confirmed by selection in plant cell
cultures. We will then insert new DNA sequences into appropriate deleted
regions and transform plant cells by selection for drug resistance. The
fate and expression of this second gene in plant cells will be studied.
Our goal is to introduce a gene for the maize storage protein, zein, into
the transformation vector and to reintroduce this gene back into maize
tissue culture cells. The ability to select for a linked marker gene will
allow us to address many questions about what happens to the zein gene in
such gene transfer experiments.
Status | Finished |
---|---|
Effective start/end date | 1/1/90 → 1/1/90 |
Funding
- National Institute of General Medical Sciences
ASJC
- Genetics
- Molecular Biology
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