Difference between revisions of "Part:BBa K731480"

Revision as of 15:28, 30 August 2012

IPTG Inducible sfGFP tagged Cysteine desulfhydrase (CysDes)

A sfGFP tagged CysDes inducible by IPTG.

Usage and Biology

CysDes is a unique 45 KDa hemolysin cysteine dependent, that was shown to have also aminostransferase activity. (1, 2) The enzyme catalyze the degradation of L-cysteine to produce hydrogen sulfide, ammonia and pyruvate. (FIGURE con pathway)

This part produces high levels of CysDes enzyme upon IPTG induction. Protein expression levels have been monitored with the sfGFP tagged composite part BBa_K731480.

Part BBa_K731400 has been fully characterized in psB1C3 and also in the low copy vector psB4K5 using E.coli strain NEB10b.

Please note that this part produces hydrogen sulfide, which is toxic if inhaled in high concentrations. Cells handling should be done under a chemical hood.

FIGURE 1. NEB10b cells transformed with BBa_K731480 were grown in LB at 37C until OD of 0.7 and induced with different concentrations of IPTG. After 4 hours a 1.5 mL aliquot was taken from each sample. Cells were span down and resuspended in 1.5 mL of PBS. Fluorescence measurements were taken with a Varian Cary Eclipse Spectrophometer using an excitation wavelenght of 464 nm.

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 2460
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1559
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 2490

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 Please note that this part produces hydrogen sulfide, which is toxic if inhaled in high concentrations. Cells handling should be done under a chemical hood.
-[[Image:Andrea2.jpg]]
+<div style="text-align:center;">[[Image:Andrea2.jpg]]</div><br/>
+'''FIGURE 1.''' NEB10b cells transformed with BBa_K731480 were grown in LB at 37C until OD of 0.7 and induced with different concentrations of IPTG. After 4 hours a 1.5 mL aliquot was taken from each sample. Cells were span down and resuspended in 1.5 mL of PBS. Fluorescence measurements were taken with a Varian Cary Eclipse Spectrophometer using an excitation wavelenght of 464 nm.