Avinash Sunder

7 Questions 11 Answers 0 Followers

Questions related from Avinash Sunder

How do I estimate the Km and Vmax values for two substrate (ordered) enzyme kinetics?

I am currently trying to characterize the kinetic behaviour of an oxidoreductase enzyme that requires NAD as the second substrate. I have two initial rate curves that correspond to v vs [A] at...

23 August 2018 5,641 11 View

Can anyone help refining a protein structure with bad geometry (but good R-factors)?

I'm currently refining an enzyme structure (data from X-ray crystallography). Resolution is 2.8 Å; model was generated using MR on a template with 62% sequence identity and Autobuild (Phenix). I'm...

09 November 2015 2,803 8 View

How can I interpret enzyme kinetic data showing apparent cooperativity and substrate inhibition?

I've got a kinetic data (see attached fig) for my enzyme that is very confusing, and I'm neither sure what it signifies nor am I able to plot it and interpret the parameters convincingly (I'm...

10 July 2014 1,504 14 View

Both recombinant and self-ligated plasmid in the same clone?

I am trying to clone my gene in pet22b (using NdeI/XhoI double digestion for 4h, gel extraction and ligation at 16 deg overnight). I got a couple of positive clones in colony PCR, when I extracted...

06 August 2013 9,146 3 View

PCR not working with similar primers

I'm encountering problems with amplifying the same gene when I use different primers - F1: GGC TAG TCT AGA ATG (TGT ACG CGG TTC GTT TAT CTG GAT C) R1: CGA GTA GAT CTC (TAG AGC CCC GCG AAT TCA AAC...

11 July 2013 2,562 3 View

Expression of periplasmic proteins

I want to clone a protein from a gram-negative bacterium that has a natural periplasmic signal peptide. Is it ok to delete the signal sequence, put the N-terminal amino acid directly next to a...

15 June 2013 9,877 1 View

How do I design primers to express a protein with C-terminal His tag?

I'm trying to clone and express a protein in E.coli using pET26b or 33b vector. I am designing primers using the NcoI and XhoI restriction enzymes. I want to know how to design primers to include...

27 May 2013 9,773 9 View