Which plasmid is used full to clone Mtb peptide and clone in M. smegmatis?

10 October 2012 0 5K Report

I want to produce some Mtb peptide in high amount and want to clone in M. smegmatis. Can anyone suggest the best plasmid for cloning and high expression?

Badges
Science topic

More Sudeep Maurya's questions See All

Is there any mouse Cell line producing CCL20/MIP3alpha?

Hello everyone Is there any mouse cell line secreting CCL20/MIP3alpha? Can we use human CCL20 in mouse for its bio-activity or not?

07 August 2016 8,915 3 View

Can any one suggest protein linkers?

Can anyone suggest a eukaryotic linker for proteins with the property of auto cleavage or protease sensitive and size 3-6 amino acids.

03 April 2016 1,998 3 View

Why recombinant gene is not getting expressed?

I am trying to express fusion peptides of mycobacterium tuberculosis in BL21DE3 and Rosseta DE3 but not getting any expression. Vector I have used is pET23a and pET28b. Gene is fusion of 6...

02 March 2016 7,385 5 View

Is it possible to clone Promoter and RBS in to an expression vector?

I want to know that is there any possibility to clone complete promoter and ribosome binding site in a expression vector to increase the cloning site. I want to clone more than one gene to express...

10 November 2014 8,180 2 View

Will the addition of OADC instead of ADC in 7H9 broth affect the mycobacterial growth?

I would like to know whether the addition of OADC instead of ADC will impact the growth of mycobacterium.

08 September 2014 5,462 4 View

How can I carry out co-expression of mammalian gene into mycobacterium?

I want to express two mammalian gene into single plasmid of mycobacterium. In which way should these gene products should get secreted and separated? So which kind of strategy I should follow and...

07 August 2014 849 3 View

Restriction Ligation

I am trying to ligate my plasmid digested with BamH l and Hind lll and same to my gene of interest but I am unable to ligate them. I have used alkaline phosphatase digestion for plasmid but still...

11 December 2013 9,817 13 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 does my protein refolded to beta sheet during thermal denaturation analysis?

Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...

06 August 2024 1,989 3 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

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

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 View

What is the relationship between protein structure and N or C terminal tagging choosing?

I want to do 2,3-butanediol dehydrogenase(BDH) enzyme purification to confirm its activity for 2,3-butanediol. Before that, I need to confirm which N or C terminal tagging is better for enzyme...

28 July 2024 366 3 View

Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...

28 July 2024 3,178 3 View

EDC carbodiimide and NHS easter 's half life among different pH?

It has been long known that EDC carbodiimide and NHS easter's half life agains hydrolysis are highly related to pH. However, very surprisingly, not yet found a paper giving a table/chart of their...

25 July 2024 8,738 0 View

Illustra™ MicroSpin™ G-25 columns what it is used for?

Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...

25 July 2024 4,927 3 View

Does post-translational protein modification cause devisions on observed pI verses calculated pI?

In running two-dimensional gel electrophoresis on bacterial protein, some spots that appear to match a protein sequence have a significantly more acidic isoelectric point than the calculated pI....

24 July 2024 8,076 3 View