Matt Gethers/OWW Syntax

OWW Syntax

The Official Syntax

Offical Syntax

My Short Cuts

Bibliography

1. Arias-Barrau E, Olivera ER, Luengo JM, Fernández C, Galán B, García JL, Díaz E, and Miñambres B. The homogentisate pathway: a central catabolic pathway involved in the degradation of L-phenylalanine, L-tyrosine, and 3-hydroxyphenylacetate in Pseudomonas putida. J Bacteriol. 2004 Aug;186(15):5062-77. DOI:10.1128/JB.186.15.5062-5077.2004 |
2. Fuangthong M, Sallabhan R, Atichartpongkul S, Rangkadilok N, Sriprang R, Satayavivad J, and Mongkolsuk S. The omlA gene is involved in multidrug resistance and its expression is inhibited by coumarins in Xanthomonas campestris pv. phaseoli. Arch Microbiol. 2008 Mar;189(3):211-8. DOI:10.1007/s00203-007-0310-1 |
3. Fuangthong M and Helmann JD. Recognition of DNA by three ferric uptake regulator (Fur) homologs in Bacillus subtilis. J Bacteriol. 2003 Nov;185(21):6348-57. DOI:10.1128/JB.185.21.6348-6357.2003 |

Files

pET-11a

Thumbnails

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Scaffold Map of High Copy Vector (Genbank Files to Follow)

Linking to Pages in OWW

The GFP and Mcherry scaffolds I'm using were constructed by Heather Keller of the Endy Lab.

Linking to Pages outside of OWW

lambda pL promoter (BBaR0011)

Tables

Approach	Pros	Cons	References
DNA (5')/MLS Conjugate	24mer oligo has been successfully imported into isolated yeast mitochondria using this method (Vestweber et al); Reagents are readily accessible and cheap.	It is unknown if the conjugate (specifically the peptide portion) can get across the yeast cell membrane; Crosslinking of DNA to peptide may inhibit expression and recombination.	[4]
DNA (Internal)/MLS Conjugate	~300 bp strand of DNA successfully imported into isolated rat mitochondria (Siebel et al).	Experiment performed with mammalian mitochondria;Crosslinking of DNA to peptide may inhibit expression and recombination.	[5]
DNA/MLS Conjugate (Crosslinking to peptide done in vivo)	Resolves problem of getting peptide across cell membrane.	Singly-bound crosslinker is likely bind any sulfhydryl group it encounters during transformation.	Natalie's Suggestion
DNA/MLS Conjugate with Cytosolic leading signal (Endocytosis)	Resolves problem of getting the conjugate across the yeast cell membrane without attempting to crosslink in vivo	Digestion or rejection during endocytosis? Formation of lipid membrane around conjugate?	Drew's Suggestion