help!!! for plasmid YIElD

Steven Goldberg goldberg at bms.com
Tue Oct 8 08:59:35 EST 1996


In article <01bbb38f$3e926f60$8c91fccc at ------>, "Takbum Ohn"
<ahnss at hitel.kol.co.kr> wrote:

> Hello!! netters...
> 
> I have cloned a cell wall hydrolase (probably toxic to E. coli host) from
> Moraxella sp. and now, I subcloned the insert as short as 1 kb. 
> 
> The subclone expressed the gene very highly  but, oddly, the copy of the
> subclone plasmid became very low. 
> 
> I don't know what make my plasmid like that.
> 
> The mother plasmid is pT7T3 19u, a high-copy number phasimid vector.
>

You said the product of your cloned gene was toxic to E. coli...so I would
guess the subclones you isolated were plasmids with mutant (low) copy
number.  E. coli probably cannot survive if expression is from a high copy
number plasmid in this instance.

Steve



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