Difference between revisions of "Part:BBa K1432001:Design"
 
Line 6: Line 6:
  
 
===Design Notes===
 
===Design Notes===
 +
We will construct a mlr promoter-GPF-mlrA cluster and clone it into the E. coli-L. lactis shuttle expression vector. So it can be easily constructed in the laboratory and could avoid secondary pollution in the natural environment of E.coli.
 +
we synthesized mlrA gene, there are 336 codons in Sphingomonas species ACM-3962. We replace 246 of them in order to increase the expression of mlrA because they are not abundant codons in Lactococcus lactis.
 +
And we successfully did it both in E.coli and L.lactis. When we induced the expression of protein, it shows that L.lactis could be more efficiently expressed than E.coli.
 +
 +
 +
The codons of Lactococcus lactis subsp. we used.
 
* TAA  
 
* TAA  
 
A GCT
 
A GCT
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===References===
 
===References===
 +
Environ Toxicol. 2001;16(6):523-34
 +
Lactococcus lactis subsp. cremoris SK11 [gbbct]: 2504 CDS's (696252 codons)
 +
http://www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi?species=272622

Latest revision as of 21:50, 2 November 2014

A Synthetic microcystin-degrading Mlr A gene


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

We will construct a mlr promoter-GPF-mlrA cluster and clone it into the E. coli-L. lactis shuttle expression vector. So it can be easily constructed in the laboratory and could avoid secondary pollution in the natural environment of E.coli. we synthesized mlrA gene, there are 336 codons in Sphingomonas species ACM-3962. We replace 246 of them in order to increase the expression of mlrA because they are not abundant codons in Lactococcus lactis. And we successfully did it both in E.coli and L.lactis. When we induced the expression of protein, it shows that L.lactis could be more efficiently expressed than E.coli.


The codons of Lactococcus lactis subsp. we used.

  • TAA

A GCT C TGT D GAT E GAA F TTT G GGA H CAT I ATT K AAA L TTA M ATG N AAT P CCA Q CAA R CGA S TCA T ACT V GTT W TGG Y TAT



Source

Sphingomonas sp. ACM-3962

References

Environ Toxicol. 2001;16(6):523-34 Lactococcus lactis subsp. cremoris SK11 [gbbct]: 2504 CDS's (696252 codons) http://www.kazusa.or.jp/codon/cgi-bin/showcodon.cgi?species=272622