Biology-Online • View topic - HFQ Protein from bacteria
Login

Join for Free!
122393 members
Advertisement
Advertisement

HFQ Protein from bacteria

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderator: BioTeam

HFQ Protein from bacteria

Postby kishsel » Thu Dec 31, 2015 7:14 am

I m currently working on Proteus mirabilis HFQ protein binding sRNA identification and characterization. I have constructed the recombinant plasmid by ligating HFQ gene from Proteus mirabilis into pET 28 recombinant vector and subsequently overexpressed the HFQ protein using IPTG induction in a competetnt BL-21 E.coli cell.. This protein was further purified by elution with an appropriate concentration of imidazole.. I have also extracted the total RNA from Proteus mirabilis using Trizol procedure.

I am ready for the Protein-RNA binding assay, but i need help in a standardized procedure to execute the Protein-RNA binding assay.

Thank you
kishsel
Garter
Garter
 
Posts: 2
Joined: Wed Dec 30, 2015 6:28 am

Postby claudepa » Thu Dec 31, 2015 4:21 pm

Hi. I am sure you will have a precise answer from a researcher using this technic. I am only curious about sRNA. I know many RNAs but not this one. I suppose that the principle of the technic after binding is the digestion of unprotected RNA by RNase. Then digestion of the protein by a protease and finally analysis of the protected RNA ? Have a good work.
claudepa
Death Adder
Death Adder
 
Posts: 68
Joined: Tue Nov 03, 2015 12:43 pm

Re: HFQ Protein from bacteria

Postby kishsel » Sat Jan 02, 2016 12:36 pm

sRNA are bacterial small non-coding RNAs. My objective is in the identification and characterization of Proteus mirabilis HFQ protein binding small non-coding RNAs.
kishsel
Garter
Garter
 
Posts: 2
Joined: Wed Dec 30, 2015 6:28 am


Re: HFQ Protein from bacteria

Postby claudepa » Sat Jan 02, 2016 7:55 pm

Thank you. I am more focused on eukaryotic cells, where there are many regulatory small RNAs. Good to know that there are also non coding RNAs in bacterias. I know that RNase protection assay can be used to identify RNA sequences for RNA binding proteins.
claudepa
Death Adder
Death Adder
 
Posts: 68
Joined: Tue Nov 03, 2015 12:43 pm


Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 1 guest