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090			_aB-18663
245	1	0	_aThe pST44 polycistronic expression system for producing protein complexes in Escherichia coli.
490	0		_vProtein expression and purification, 40(2), p.385-395, 2005
520	3		_aProtein complexes are responsible for key biological processes, but methods to produce recombinant protein complexes for biochemical and biophysical studies are limited. We have developed a second generation Escherichia coli polycistronic expression system which improves on the modularity of our original pST39 polycistronic system. This pST44 expression system simplifies the construction of polycis-tronic plasmids, particularly of variant plasmids expressing deletion or point mutations in any subunit. To facilitate purification of the expressed complex, we have prepared a suite of 72 plasmids which allows individual subunits to be tagged at the N- or C-terminus with six permanent or cleavable peptide affinity tags. We demonstrate these new features in a detailed deletion analysis of a three protein yeast Piccolo NuA4 histone acetyltransferase complex, and in the affinity purification of a human Piccolo NuA4 complex. We also utilize the modular design to show that the order of expression of the three subunits along the polycistronic plasmid does not affect the reconstitution of the yeast Piccolo complex in E. coli.
650	1	4	_aPOLYCISTRONIC EXPRESSION
650	1	4	_aCOEXPRESSION OF PROTEIN COMPLEXES
650	1	4	_aAFFINITY TAGS
650	1	4	_aHISTONE ACETYLTRANSFERASE
650	1	4	_aRECOMBINANT EXPRESSION
700	1	2	_aTan, S.
700	1	2	_aKern, R. C.
700	1	2	_aSelleck, W.
856	4	0	_uhttps://drive.google.com/file/d/1Kc7Hf3-z7gb1HgFlVxbrmJVWsZ3-YEaN/view?usp=drivesdk _zPara ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx
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