Loss of expression, help!

[Dr. Duncan Clark]

In article <NDBBKMADGLKJLODKIFGAEEOMCAAA.wind at biobase.dk>, the eminent
Troels Wind at BIOSCI/MRC Human Genome Mapping Project Resource Centre
wrote
>It is not a question of codon-choice; all codons are selected for optimal
>coli-expression.

Secondary structure i.e. big hairpin in the mRNA of the second
construct.

One of the additional sequences makes your protein lethal? 

The second construct is re-arranging or kicking the plasmid out as it
grows.

I presume sequencing shows the 2nd construct is correct with no
accidental change of reading frame etc. 

Duncan

 
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
GeneSys Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk