We are in the process of designing a new version of the screening plasmid to account for some of the shortcomings of the previous version. An early list of possible improvements:
We are in the process of designing a new version of the screening plasmid to account for some of the shortcomings of the previous version. Details can be found here:
#*[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16269719&query_hl=2&itool=pubmed_docsum Propionate-inducible] induction system, has a larger dynamic range than pBAD and doesn't require a specialized strain. (i.e. no knockouts of native systems needed.)
#**[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16333863&itool=pubmed_DocSum more details of this system]
#It looks like MC4100 is missing the propionate metabolism genes and so wouldn't work with this system, these are the two genes in K12 are homologous to the prpBCDE operon:
#*[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=49175990&db=Nucleotide&from=349236&to=350405&view=gbwithparts 349236..350405], [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?val=49175990&db=Nucleotide&from=350439&to=351890&view=gbwithparts 350439..351890] Unfortunately, there is a [http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=12618467 98kb deletion] from the MC4100 (a K12 derivative) genome from 274723-371962, so it's missing these genes.
#*However, I'm not tied to MC4100, would rather get this working in K12 and then we could maybe make some progress on doing directed deletions of K12 as planned in the [[Standard E. coli Strain for BioBricks|Biobricks Standard Strain]].
==References==
==References==
Revision as of 14:04, 30 May 2006
This page is a work in progress.
Introduction
The screening plasmid is designed to enable fast characterization of PoPS-based parts and devices. It consists of 4 components: (1) Tunable input (2) Input measurement (3) Part/Device insertion site (4) Output measurement.
We are in the process of designing a new version of the screening plasmid to account for some of the shortcomings of the previous version. Details can be found here:
[2] Khlebnikov et al,Modulation of gene expression from the arabinose-inducible araBAD promoter. J Ind Microbiol Biotechnol. 2002 Jul;29(1):34-7.
[3] Effect of gene location, mRNA secondary structures, and RNase sites on expression of two genes in an engineered operon. Biotechnol Bioeng. 2002 Dec 30;80(7):762-76.