Giggsy Matinny

6 Questions 2 Answers 0 Followers

Questions related from Giggsy Matinny

E. coli strain Lemo21(DE3) can express transmembrane protein in secretion form?

My recombinant protein is transmembrane protein that cloned into pET-22b that contain PelB signaling peptide at the N-terminal. I need to express recombinant protein in a secretion form that...

23 June 2016 4,212 1 View

Question about plasmid digested by two-cut-site RE at MSC?

Here I have a basic question. My cutting vector run faster than uncut when cut with MscI and XhoI. Who can tell me it could happen in the normal situation?

13 February 2016 430 1 View

Which is the optimal OD for protein induction?

My protein is expressed at 20C for 18h with 0.1mM IPTG Anyone can suggest optimal OD for induction?

08 November 2015 2,708 5 View

Can anyone help with my problem with periplasmic protein extraction by osmotic shock?

I'm very confuse about time for incubation of protein after add sucrose buffer and osmotic shock buffer(MgSO4). and volume of each buffer were added?

08 November 2015 1,312 1 View

How do I express a secreted protein at 30-37’C?

What is the optimal IPTG concentration and incubation time after induction? Normally, My protein were induced with 0.5mM at 25'C for O/N but now I have limitation about shaker, so i'll to try new...

22 September 2015 9,747 3 View

Why is my protein not exported to periplasmic space of E. coli?

My protein is fused with signal peptide of E. coli and I'm express in BL21(DE3) at 37'C for 3hr after added 1mM IPTG. Can anyone suggest Why my protein can not export?

19 June 2015 3,129 4 View