Autoinduction of pGEX5x3?

09 September 2017 0 6K Report

Hello everyone,

I wish to express a gene cloned in pGEX5x3 through auto-induction and compare with the normal IPTG induction. Any one has experience doing autoinduction with pGEX vectors?

Badges
Science topic

Similar topics
Vectorization

More Robert Adamu Shey's questions See All

How do I purify rabbit IgG4?

I want to purify IgG4 antibodies from immunized rabbits. I have tried to look for protocols to no avail. Commercial kits on the other hand exist for humans and a few other animals but I have not...

03 April 2019 10,153 3 View

Any bioinformatics tools to predict immuno-modulatory or immuno-regulatory epitopes in proteins?

Hello everyone, we are characterizing some proteins in the context of vaccine development and we are interested in determining there are any epitopes that may rather lead to immune evasion. Don't...

03 April 2019 3,080 3 View

How do I avoid impurities from my SEC purification?

Hello everyone, so I expressed this protein and I am trying to further purify using SEC and still having the impurities in the fraction I expected to have the protein. How do I eliminate these. I...

03 April 2019 7,714 3 View

How do i prepare water saturated phenol:chloroform mixture (5:1)?

I am doing RNA, DNA and protein extraction from the same sample and my protocol recommends this water saturated phenol:chloroform mixture. I am not not very versed with how to go about with the...

03 April 2018 4,443 8 View

What are the best (recommended) tools for 3D protein structure prediction, refinement, and validation; also what is the best tool for MD simulation?

Hello everyone, I am trying to do some hands-on bioinformatics and will be grateful to get your orientation.

31 December 2017 1,479 5 View

How do I interpret protein 3D structure refinement data?

Hello everyone, I did 3D structure prediction (using I-TASSER) for a protein and submitted the best model for structure refinement (with GalaxyRefine). Unfortunately, I don't have a very good...

11 December 2017 1,362 0 View

What is the best buffer to us in antigen-capture ELISAs?

Hello everyone, I am about doing an antigen capture ELISA and a little confused about which buffer can be used in absorbing the capture antibody to the microplates. I am using Nunc MaxiSorb 96...

10 November 2017 2,991 3 View

What is the effects of coating ELISA plates with antigen for longer than recommended durations?

Hello everyone, So I planned this ELISA to do, unfortunately I coated plates and for some reason I am not able to continue with the experiment as planned. So basically, instead of coating...

09 October 2017 9,378 4 View

Isolation of inclusion bodies?

I am working with a pET30a+ construct and my protein is C-terminal His-tagged. My protein is forming inclusion bodies and I need to get a protocol for isolation of a pure protein from this. An...

07 August 2017 2,097 4 View

Signal peptides in secretory proteins?

I wish to find out if all secreted proteins have a signal peptide. Also, if yes, are these signal peptides always cut?

07 August 2017 10,016 2 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Why did the authors extrapolate a phenotype that they experimentally proved in one bacterial strain across the whole genus of the organism?

I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...

05 August 2024 6,787 4 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

How to load and plot 2D-PIV (particle image velocimetry) recorded velocity vector field in Techplot?

Dear Researchers I need to know, how to load and plot 2D-PIV (particle image velocimetry) recorded velocity vector field data in Tecplot? Thank You

02 August 2024 3,615 1 View

How can we now if the promotor in the vector is well?

Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...

02 August 2024 3,987 1 View

What is the acceptable p-value cutoff for GO enrichment analysis ?

I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...

31 July 2024 225 2 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

Inquiry on Maximum Nucleic Acid Volume for 2.5 mL Liposome Solution?

I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...

30 July 2024 6,420 1 View

How are aPTT and PT values reduced after hemodialysis compared to before dialysis value in patients with chronic kidney disease?

I am conducting my research regarding blood coagulation in patients with CKD. I observed a reduction in post hemodialysis aPTT and PT values compared to pre hemodialysis values. But most of...

30 July 2024 3,467 0 View

Are these cassettes suitable for expressing PETase mutant in E. coli?

I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...

28 July 2024 7,559 1 View