Dear Dr. artem
thank you for your suggestion. But I have done lots of transformations
with E.coli as the host vector for my different constructs with the same
plasmids, and I take care to use strains which are all dcm-, dam-. I never
had any problem in this regard. We have also standardised protocols for
various other Bacillus species which works out on a regular basis. But here
the problem is I am not sure about the type of strain it is, even on a
16SrRNA analysis and visual examination, it does look like a strain of B.
subtilis. But the natural competency method did not work out.
regards
jayakumar
----- Original Message -----
From: "Dr. Artem Evdokimov" <eudokima at mail.ncifcrf.gov>
To: <microbio at hgmp.mrc.ac.uk>
Sent: Monday, June 11, 2001 1:05 AM
Subject: Re: transformation of Bacillus
> Are you sure that your DNA is compatible ? What I mean is - perhaps it
> is so toxic that all the transformed cells croak ?
>> Have you tried transforming your DNA into other bacteria, just as a
> control ?
>> Just a thought.
>> A.
>> "R. Jayakumar" wrote:
> >
> > hi..
> > I badly need some help here. I have been trying my best to transform
my
> > bacillus isolate with a plasmid construct. The isolate looks like a new
> > strain of Bacillus subtilis. But I need to put my construct into it. I
> > have tried natural competency method, electroporation etc..and even
> > triparental mating. But to no avail. The strain stubbornly refuses to
> > accept the DNA> Can any good samaritan give me some ideas on how to go
> > about it. I will be eternally grateful to him.
> > thank you
> > sincerley
> > jayakumar
> >
> >
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> > \\\\\\\\\\\\\\\\\\\\\\\\\\\
> > R. Jayakumar
> > Dept. of Molecular Microbiology,
> > School of Biotechnology,
> > Madurai Kamaraj University,
> > Madurai - 625021. India.
> > tel: +91-452-859141(night hours), +91-452-858471 (ask to connect to 374)
> > fax:+91-452-859105
> > Efax: (603)-688-4665
> > Email: jakkku at mrna.tn.nic.in, (send a copy to jakku at zxmail.com ....just
in
> > case)
> >
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> > \\\\\\\\\\\\\\\\\\\\\\\\\\\
> > Science is constructed of facts like a house is of bricks, but a mere
> > accumulation of facts does not make science nor does a pile of bricks a
> > house
> > -Henri Poincare
> >
> > ---
>> --
> |Dr. Artem Evdokimov Protein Engineering |
> | NCI-Frederick Tel. (301)846-5401 |
> | FAX (301)846-7148 |
> | eudokima at mail.ncifcrf.gov |
> | http://www.ncifcrf.gov/plague |
>>
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