Sinopsis
Molecular biology, in particular recombinant DNA research, has transformed research in the biological and medical sciences. This technology currently influences all aspects of biological research, has far-reaching applications in clinical diagnosis, and has led to important developments in agriculture and biotechnology. This course provides a hands-on introduction to molecular biological methods, including molecular cloning, polymerase chain reaction (PCR), Southern (DNA) blotting, Northern (RNA) blotting, DNA sequencing, oligonucleotide-directed mutagenesis, and protein expression, purification, and detection. You will work with a well-characterized gene (virD2) from the Agrobacterium tumefaciens tumor-inducing (Ti) plasmid. Virulence (v/r) genes mediate transfer of a specific region of the Ti plasmid from A. tumefaciens into host plant cells; oncogenes contained in the transferred DNA integrate into the host nuclear genome where their expression causes tumorous growth. Because we have studied virD2 previously, both starting materials and finished products for each experiment are available. All of the experiments you will do are "real," and most are published. This approach will demonstrate practical aspects of experimental design. Although the gene you will use is bacterial, the techniques apply to any research system.
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