Difference between revisions of "Part:BBa K200005"

(References)
Line 21: Line 21:
 
<partinfo>BBa_K200005 parameters</partinfo>
 
<partinfo>BBa_K200005 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
===Design Notes===
 +
PCR from Escherichia Coli BL21(DE3)
 +
 +
Primers Used:<br>
 +
Forward: <br>
 +
<b>GCTCTAG</b>ATGACAGAACCGTTAACCG <br>
 +
Recomended Temperatures for PCR : 54.5C(without overhang) and 64.8C(with overhang)
 +
<br><br>
 +
Reverse:<br>
 +
<b>GGACTAGTA</b>TTAGATACTACGACTAAACGAC<br>
 +
Recomended Temperatures for PCR : 54.7C(without overhang) and 64.2C(with overhang)<br><br>
 +
<i>Biobrick Overhang shown in Bold</i>
  
 
===References===
 
===References===
 
<biblio>#otsa1 pmid=12105274 </biblio>
 
<biblio>#otsa1 pmid=12105274 </biblio>

Revision as of 15:19, 1 September 2009

OtsA: Part 1 of 2 for trehalose producing enzymes.

Sequence codes for trehalose-6-phosphate synthase enzyme. This enzyme is the first of two required in the conversion of glucose to trehalose.
This enzyme catalyses the following reaction:
UDP-glucose + D-glucose 6-phosphate -> UDP + alpha,alpha-trehalose 6-phosphate

Trehalose is a disaccharide formed from two glucose molecules. Throughout nature, trehalose is associated with resistance to dessication and cold shock otsa1, and is naturally produced in Escherichia Coli. We hope that by upregulating the trehalose production pathways in E.coli we can increase trehalose concentrations within our cell, thereby conferring some resistance to protein degredation in our system. This would allow easy transport and storage of the final product.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 383
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

PCR from Escherichia Coli BL21(DE3)

Primers Used:
Forward:
GCTCTAGATGACAGAACCGTTAACCG
Recomended Temperatures for PCR : 54.5C(without overhang) and 64.8C(with overhang)

Reverse:
GGACTAGTATTAGATACTACGACTAAACGAC
Recomended Temperatures for PCR : 54.7C(without overhang) and 64.2C(with overhang)

Biobrick Overhang shown in Bold

References

<biblio>#otsa1 pmid=12105274 </biblio>