production of protein in yeast

Frank Ho hof at chiremv.chiron.com
Wed May 8 19:00:12 EST 1996


tliang at utmmg.med.uth.tmc.edu (T. Chyau Liang) writes:

>In article <3180C597.7E69 at rp.fr>, jean-dominique.guitton at rp.fr wrote:

>> We try to produce a recombinant protein intracellularly in Yeast. 
>> After cell lysis we are not able to detect the protein by SDS PAGE 
>> using classical denaturation process in the SDS beta-mercaptoethanol 
>> denaturation buffer. If we want to observe something we need first to 
>> heat the lysate at 60°C for 30mn then to add the denaturing buffer. 
>......

>I don't know how you visualize your protein after SDS.  If you used
>western blot and you could not see your protein until you heated it with
>denaturants, we have seen a similar phenomenon from insect cells due to
>the conformational dependence of the epitope. Please see Arch. Biochem.
>Biophys. 305(2), 563-569, 1993.

>Hope this helps.

>T. Chyau Liang
>U Texas - Houston



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