Difference between revisions of "Part:BBa K1949020:Design"

(Material and method)
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yafN gene fragment amplified by PCR contains XbaI site at N-terminal and SpeI and PstI site at C-terminal. Both terminals were cut by respective restriction enzymes and the fragment used for ligation with iGEM plasmid vectors.
 
yafN gene fragment amplified by PCR contains XbaI site at N-terminal and SpeI and PstI site at C-terminal. Both terminals were cut by respective restriction enzymes and the fragment used for ligation with iGEM plasmid vectors.
 
 
===Material and method===
 
 
  
  

Revision as of 01:53, 16 October 2016


yafN


Design Notes

yafN(BBa_K1949020) was gained by PCR using E. coli MG1655 genomic DNA and the following primers.

fwd:5’-aaatctagatgcatcgaattctcgctgaaaaatcg-3’

rev:5’-cccctgcagcggccgctactagtattattattccttaaagtcattgaaatcatcatccgtc-3’

yafN gene fragment amplified by PCR contains XbaI site at N-terminal and SpeI and PstI site at C-terminal. Both terminals were cut by respective restriction enzymes and the fragment used for ligation with iGEM plasmid vectors.


Source

From E.coli MG1655.

Material and method

References

[1] Zhang Y, Yamaguchi Y, Inoue M. Characterization of YafO, an Escherichia coli Toxin. J Biol Chem. 2009 Sep; 284(38): 25522–25531.

[2] Dalsgaard M, Jørgensen M, Gerdes K. Three new RelE-homologous mRNA interferases of Escherichia coli differentially induced by environmental stresses. Mol Microbiol. 2010 Jan; 75(2): 333–348.

[3] Singletary LA, Gibson JL, Tanner EJ, McKenzie GJ, Lee PL, Gonzalez C, et all. An SOS-Regulated Type 2 Toxin-Antitoxin System. J Bacteriol. 2009 Dec;191(24):7456-65.