tributed, no over
> representation of any mutation. Isolated wild type
> clones generate mutants after transformation.
>
> Chun
>
>
>
> From: CCP4 bulletin board
> [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of RubA(c)n
> SA!nchez Eugenia
> Sent: Wed
Sánchez Eugenia
Sent: Wednesday, January 25, 2012 8:52 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] Expression of Viral proteins for crystallography
Dear Gregory and Darren,
Thank you for your answers. They have been very useful.
2012/1/24 Gregory Verdon
it possible that the protein is
Dear Gregory and Darren,
Thank you for your answers. They have been very useful.
2012/1/24 Gregory Verdon
>
> it possible that the protein is toxic (even when slightly expressed from
> your possibly leaky pET vector), so that e.coli select for mutations that
> kill expression of your recombinant
Rubén,
the previous answer probably addresses your problem accurately.
However. If your protein of interest modifies DNA/RNA, it is quite common that
your pET constructs will mutate rapidly in E.coli. Lac operons tend to leak
quite a bit, which is not enough to detect the protein prior to indu
Inspired by the recent post about "quasispecies:"
I have been bothered recently by the following problem: why do species
of genetic uniformity exist at all (or do they?)? This first came up
when I saw a Nature paper describing live bacteria extracted from a
supposedly 250-million-year-old salt cry
I think the explanation is this:
The source is natural viral RNA which is a mixture of naturally mutated
sequences (e.g. flu forms such a quasispecies)
See:
http://www.virology.ws/2009/05/11/the-quasispecies-concept/
The pooled RNA has an average sequence that you see when you sequence the
pooled
Dear everyone,
I am trying to clone a viral protein in the E. Coli BSJ strain and i am
having some problems.
I start from the viral RNA carrying out a reverse transcription and PCR
(RT-PCR) to obtain the protein cDNA. When I sequence this cDNA to check for
mutations, there are no mutations. So th
