I need to knock out one of the gene of my interest from the lytic bacteriophage. I am presently thinking of homologous recombination but wondering whether it is feasible or not.
It depends on the particular lytic bacteriophage you are working with. For example, bacteriophage T4 has glycosylated, hydroxymethylcytosine in its DNA to evade the host defense mechanisms. Also, T4 makes many gene products that degrade the host DNA, so the chance of using homologous recombination to knock out a phage T4 gene is low.
You also have to consider how you will select for your phage that have your gene of interest knocked out. If you have a convenient phenotype to score, it is a lot easier. If you have to rely on screening individual plaques by PCR it will be possible, but not easy.
Historically, researchers used transposons to knock out phage genes, and you could try something similar.
Thank you David. T4 is my phage of interest. Can something other than homologous recombination be tried for this phage?
As long as you have a way of selecting for your mutation of interest, you can use any of the normal mutagens: EMS, MMS, acridines, etc. Here is a link:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1201963/
I also found this vector system that may work for you:
http://www.jbc.org/content/263/23/11336.full.pdf
You may be able to do homologous recombination after all.
Here is a nice collection of T4 references:
http://www.mansfield.ohio-state.edu/~sabedon/bib_t4_book.htm
Good luck!
Thanks a lot David. All the links are of great help.
I also came across Insertion/substitution vector system. Most probably i'll design something similar for my work.
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