https://eln.excellgen.com/index.php?action=history&feed=atom&title=PKA_alpha_expression_with_Chaperones PKA alpha expression with Chaperones - Revision history 2026-10-18T07:09:11Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=PKA_alpha_expression_with_Chaperones&diff=16636&oldid=prev Paul.xu at 22:09, 20 November 2017 2017-11-20T22:09:47Z <p></p> <p><b>New page</b></p><div>Medium: LB/Kan/Chl<br /> <br /> Screening of PKA alpha expression with chaperones<br /> <br /> [[File:Chaperone.jpg|RTENOTITLE]]<br /> <br /> {| cellspacing=&quot;0&quot; border=&quot;2&quot; rules=&quot;NONE&quot; frame=&quot;VOID&quot;<br /> |-<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | #<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 63px;&quot; | Plasmid<br /> | width=&quot;150&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 132px;&quot; | Chaperone<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 74px;&quot; | Promoter<br /> | width=&quot;120&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 98px;&quot; | Inducer<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | Resistant Marker<br /> |-<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | 1<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 63px;&quot; | pG-KJE8<br /> | width=&quot;150&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 132px;&quot; | dnaK-dnaJ-grpE<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 74px;&quot; | araB<br /> | width=&quot;120&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 98px;&quot; | L-Arabinose<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | Cm<br /> |-<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | &lt;br/&gt;<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 63px;&quot; | &lt;br/&gt;<br /> | width=&quot;150&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 132px;&quot; | groES-groEL<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 74px;&quot; | Pzt1<br /> | width=&quot;120&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 98px;&quot; | Tetracyclin<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | &lt;br/&gt;<br /> |-<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | 2<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 63px;&quot; | pGro7<br /> | width=&quot;150&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 132px;&quot; | groES-groEL<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 74px;&quot; | araB<br /> | width=&quot;120&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 98px;&quot; | L-Arabinose<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | Cm<br /> |-<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | 3<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 63px;&quot; | pKJE7<br /> | width=&quot;150&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 132px;&quot; | dnaK-dnaJ-grpE<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 74px;&quot; | araB<br /> | width=&quot;120&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 98px;&quot; | L-Arabinose<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | Cm<br /> |-<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | 4<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 63px;&quot; | pG-Tf2<br /> | width=&quot;150&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 132px;&quot; | groES-groEL-tig<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 74px;&quot; | Pzt1<br /> | width=&quot;120&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 98px;&quot; | Tetracyclin<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | Cm<br /> |-<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | 5<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 63px;&quot; | pTf16<br /> | width=&quot;150&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 132px;&quot; | tig<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 74px;&quot; | araB<br /> | width=&quot;120&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; style=&quot;width: 98px;&quot; | L-Arabinose<br /> | width=&quot;30&quot; height=&quot;17&quot; bgcolor=&quot;#ffff99&quot; | Cm<br /> |}<br /> <br /> <br /> <br /> Should induce with 0.02% to 0.2% L-Arabinose (and + Tetracyclin )<br /> <br /> weight of wet cell pellet from 1 L culture<br /> <br /> {| width=&quot;200&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | &lt;br/&gt;<br /> | pellet wt<br /> | PKA yeild<br /> |-<br /> | pG-KJE8&lt;br/&gt;<br /> | 4.3<br /> | &lt;br/&gt;<br /> |-<br /> | pGro7&lt;br/&gt;<br /> | 3.9<br /> | &lt;br/&gt;<br /> |-<br /> | pKJE7&lt;br/&gt;<br /> | 3.6<br /> | &lt;br/&gt;<br /> |-<br /> | pG-Tf2&lt;br/&gt;<br /> | 1.4<br /> | &lt;br/&gt;<br /> |-<br /> | pTf16&lt;br/&gt;<br /> | 4.6<br /> | &lt;br/&gt;<br /> |}<br /> <br /> {| width=&quot;200&quot; cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | 2012/11/28&lt;br/&gt;<br /> | pellet wt from 2 L<br /> | PKA yeild<br /> |-<br /> | pG-KJE8&lt;br/&gt;<br /> | 4.0<br /> | &lt;br/&gt;<br /> |-<br /> | pGro7&lt;br/&gt;<br /> | 5.1<br /> | &lt;br/&gt;<br /> |-<br /> | pKJE7&lt;br/&gt;<br /> | 4.5<br /> | &lt;br/&gt;<br /> |}<br /> <br /> <br /> <br /> Expression screening<br /> <br /> 1&amp;nbsp; pTf16<br /> <br /> 2&amp;nbsp; pG-KJE8<br /> <br /> 3&amp;nbsp; pGro7<br /> <br /> 4&amp;nbsp; pG-Tf2<br /> <br /> 5&amp;nbsp; pKJE7<br /> <br /> 6&amp;nbsp; pTf16<br /> <br /> 7 pG-KJE8<br /> <br /> 8 pGro7&amp;nbsp;<br /> <br /> 9 pG-Tf2<br /> <br /> 10 pKJE7<br /> <br /> [[File:PKA-alpha expression screening.jpg|RTENOTITLE]]<br /> <br /> Looks like pG-KJE8 (tet+L-arabinose induction) has better expression<br /> <br /> --[[User:Ruili.su|Ruili.su]] 17:45, 18 March 2011 (EST)<br /> <br /> [[File:PKA alpha gel filtration 4 1 2011.jpg|RTENOTITLE]]<br /> <br /> [[File:PKA alpha gel filtration 4 1 2011 2.jpg|RTENOTITLE]]<br /> <br /> PKA purification using gel-filtration column<br /> [[Category:Protein Expression]]</div> Paul.xu