How to target a gene for deletion - (May/19/2008 )
Hello. My question is how to target a gene for deletion. I'm a first year graduate student in parasitology who has very little background on molecular biology. I am interested in determining the role of a gene in the parasite I am studying. I would appreciate it, if anyone could direct me to a website that could familiarize me with this topic. Thank you very much.
I would be damned. I wouldn't even know where to begin!
Gene targeting strategies are quite varied in different organisms (Bacteria, Yeast, Mouse etc), and can be done generally by PCR or homologous recombination, but there are many flavors there. And then you have random integration, gene knock-down, gene-trap, chemical mutagenesis etc too. Parasite? that I suspect would be some very specialized method.
May be I would start with Wikipedia and some review articles in your field. Any molecular biology textbook in your lab or library should provide basic idea. Definitely start there. There is no pain without gain, reverse that.
Hello, genetic manipulation on parasites depends strongly on the parasite you are working with, and what kind of gene you want to knock down.
For protozoa there are a number of transfection techniques you could use, and parasites like malaria, leishmaina, trypanosomes and toxoplasma have been successfully transfected for overexpression or knock down of genes. On the other hand transfection of worms is harder, and although there are plenty of successful techniques for C. elegans, not all of them are applicable for worms with more complicated life cycles.
Best place to start looking will be parasitology journals like, molecular and biochemical parasitology, experimental parasitology, international journal for parasitology, trends in parasitology...
Hope this helps.