Hi all,
I have a question that I cant get my head around. When you make a lambda phage library from gDNA (environmental samples) How is it that you can screen for function on specific substrates etc? Surely RNA needs to be transcribed and then translated into protein in order for functional characterization to place? Is this what happens?
thanks for your time
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lambda phage library
Started by proteinz, Oct 17 2010 03:23 AM
4 replies to this topic
#2
pDNA
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Posted 17 October 2010 - 03:32 AM
this is what we call phage display ...that allows you to connect the genetic information with the functionality of proteins!
Regards,
p
Regards,
p
#3
proteinz
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Posted 17 October 2010 - 09:10 AM
Thanks for the reply! I'm really grateful! But still a bit confused. Is this what it means?
gDNA extracted
ligated into lambda zap express vector
packaged into phage heads
titered- infect e. coli
amplified to achieve a working stock
-then simply plate onto selective plates- say plates containing starch if looking for amylases etc? As the bacteriophage produces the protein on the surface of the phage as a fusion protein?
i really appreciate your time!
gDNA extracted
ligated into lambda zap express vector
packaged into phage heads
titered- infect e. coli
amplified to achieve a working stock
-then simply plate onto selective plates- say plates containing starch if looking for amylases etc? As the bacteriophage produces the protein on the surface of the phage as a fusion protein?
i really appreciate your time!
#4
pDNA
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Posted 18 October 2010 - 09:02 AM
i dont think it will work the way you described it ...this would be far to easy.
you can read this ...they did what you are interested in.
Regards,
p
you can read this ...they did what you are interested in.
Regards,
p
proteinz, on 17 October 2010 - 09:10 AM, said:
Thanks for the reply! I'm really grateful! But still a bit confused. Is this what it means?
gDNA extracted
ligated into lambda zap express vector
packaged into phage heads
titered- infect e. coli
amplified to achieve a working stock
-then simply plate onto selective plates- say plates containing starch if looking for amylases etc? As the bacteriophage produces the protein on the surface of the phage as a fusion protein?
i really appreciate your time!
gDNA extracted
ligated into lambda zap express vector
packaged into phage heads
titered- infect e. coli
amplified to achieve a working stock
-then simply plate onto selective plates- say plates containing starch if looking for amylases etc? As the bacteriophage produces the protein on the surface of the phage as a fusion protein?
i really appreciate your time!
#5
proteinz
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Posted 19 October 2010 - 12:35 AM
pDNA, on 18 October 2010 - 09:02 AM, said:
i dont think it will work the way you described it ...this would be far to easy.
you can read this ...they did what you are interested in.
Regards,
p
you can read this ...they did what you are interested in.
Regards,
p
proteinz, on 17 October 2010 - 09:10 AM, said:
Thanks for the reply! I'm really grateful! But still a bit confused. Is this what it means?
gDNA extracted
ligated into lambda zap express vector
packaged into phage heads
titered- infect e. coli
amplified to achieve a working stock
-then simply plate onto selective plates- say plates containing starch if looking for amylases etc? As the bacteriophage produces the protein on the surface of the phage as a fusion protein?
i really appreciate your time!
gDNA extracted
ligated into lambda zap express vector
packaged into phage heads
titered- infect e. coli
amplified to achieve a working stock
-then simply plate onto selective plates- say plates containing starch if looking for amylases etc? As the bacteriophage produces the protein on the surface of the phage as a fusion protein?
i really appreciate your time!
