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Advice in microbiology needed

Scott Coutts scott.coutts at med.monash.edu.au
Fri May 24 07:55:03 EST 2002


Yes, these are definitely Cm resistant. They have the cat gene on the 
Hte plasmid that contains the extra tRNA genes. They're definitley NOT 
Ap resistant, unless you're maintaining some other plasmid with which 
they have been transformed.

I cant comment on the nutritional requirements with any authority, but 
nothing in their documentation suggests that they should require 
anything special. I thought they were standards BL21s with an extra 
plasmid (containing the tRNA genes).


Michael Witty wrote:

> On 22 May 2002, Vince Mulholland wrote:
> 
> 
>>Vladimir,
>>
>>Use chloramphenicol NOT ampicillin (the bacterium is Cam-R). This seems to
>>be your problem.
>>
>>Hi,
>>
>>BL21 CodonPlus (DE3) RIL from Stratagene has genotype E. coli B F(-) ompT
>>hsdS(rB(-) mB(-)) dcm(+) Tet(r) gal ?(DE3) endA Hte[argU ileY leuW Cam(r)],
>>and appears it should be able to grow in a minimal medium. We tried to grow
>>it in M9 with glucose (and ampicillin for plasmid maintenance) with no
>>avail. Tech. rep. from Stratagene was not of much help suggesting to add
>>proline (?), cysteine (?) and thiamine (?) with no guarantee to work. Any
>>advice what may be a problem and how to make it grow?
>>
>>Thanks,
>>
>>Vladimir Yamshchikov
>>
> 
> 
> Vince, are you sure?  Vladimair, what about the idea of spreading a
> minimal plate with cells and putting a grain or two of all the amino
> acids on your chemical shelf in sectors and see what makes a difference
> (classical experiment).  Regards, Mike.
> 
> 


-- 
Scott J. Coutts
------------------------------------------------------------
Bacterial Pathogenesis Research Group
Monash University, Australia
Phone: +61 3 9905 4838
Email: scott.coutts at med.monash.edu.au
------------------------------------------------------------




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