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Histone like protein extraction from Gram positive - (Nov/16/2005 )

I want to extract HU(Histone like protein) from a gram positive bacetria. I have tried a protocol with PCA 5% but I have had very little protein.
I am looking for a good protocol specially for the lysis of the cell wall.
I really need this protocol and will welcome any answers.
Nikeh rolleyes.gif


I work with S aureus.

I have a chromosomal prep protocol with a step for lysis of the gram positive cell wall, but I don't know if your proteins are recoverable after this step?

basically, you grow up some cells....spin, resuspend in TEN....add some lysostaphin, cook at 37C for a bit....add some SDS and some proteinase K and cook a bit longer....then extract the DNA from the proteins and cell-wall components

could you use the protocol without the proteinase K? I don't know if the rest of the protocol would hurt your protein, but it is a quick and easy way to crack the cells open. let me know if you want any details.

when we look at S aureus proteins, we clone them up and express them in e coli, so that is all I know. let me know if you want any details but it may not be useful for you.