Anna Edleson Posted July 24, 2018 Posted July 24, 2018 (edited) Hi! I am trying to conduct an experiment in which I will need to increase the gene expression of specific heat shock proteins (DnaK, HtpG, and GroEl) in E. Coli bacteria. What is the best method to do this? Edited July 24, 2018 by Anna Edleson accidentally omitted a word
CharonY Posted July 26, 2018 Posted July 26, 2018 Generally you would use whatever protocols are established in your lab, or what your supervisor suggests you to do. There are a variety of options and suitability is determined by the precise question of your project. Expression can be manipulated by manipulating cis-or trans-regulatory elements, have them in (overexpression)-vectors, make inducible or not etc. The question is also whether you just want pure gene expression or whether you also want the protein numbers to increase (both are not the same).
StringJunky Posted July 26, 2018 Posted July 26, 2018 (edited) 24 minutes ago, CharonY said: ... The question is also whether you just want pure gene expression or whether you also want the protein numbers to increase (both are not the same). Does it mean increasing the elements that create the protein as well as actually increasing that protein, depending on context? Edited July 26, 2018 by StringJunky
CharonY Posted July 26, 2018 Posted July 26, 2018 15 minutes ago, StringJunky said: Does it mean increasing the elements that create the protein as well as actually increasing that protein, depending on context? Not really. Gene expression commonly refers to transcription of genes. While the transcript levels will rise, it does not necessary result in significant translation rate. And even if it does the, increase may not be biologically relevant. There can be several layers of regulation that make direct correlation between gene expression and protein levels quite difficult.
StringJunky Posted July 26, 2018 Posted July 26, 2018 1 minute ago, CharonY said: Not really. Gene expression commonly refers to transcription of genes. While the transcript levels will rise, it does not necessary result in significant translation rate. And even if it does the, increase may not be biologically relevant. There can be several layers of regulation that make direct correlation between gene expression and protein levels quite difficult. Right. Cheers.
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