Part:BBa_K231001
Phytochelatin Synthase, HA-tag
Codes for phytochelatin synthase (PCS) which catalyzes the conversion of glutathione (GSH) to phytochelatin (PC). HA Tag appended after a TG scar/linker to enhance protein detection ability. The precursors for PC are naturally present in E. coli and have been shown to be upregulated in the presence of PCS and heavy metals (Li, Dhankher, Carriera. 2004). PC is a cysteine rich molecule capable of binding heavy metals. The PCS gene is endogenous to A. thaliana.
Growth of E. coli was not affected by insertion of this gene or induction by ITPG. Culture conditions consisted of shaking at 37 degrees in Lauria Broth.
Phytochelatin, the result of the reaction catalyzed by PCS binds arsenic. The activity assay was run to test the ability of PCS expressing E. coli to absorb arsenic from liquid broth. A time series of samples were taken from samples growing in arsenic. The bacteria was removed from the bacteria by centrifugation and arsenic levels in the supernatant were measured. Arsenic levels are measured by HPLC.
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Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1
Illegal PstI site found at 100
Illegal PstI site found at 1363
Illegal PstI site found at 1396
Illegal PstI site found at 1507 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal PstI site found at 100
Illegal PstI site found at 1363
Illegal PstI site found at 1396
Illegal PstI site found at 1507 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1
Illegal PstI site found at 100
Illegal PstI site found at 1363
Illegal PstI site found at 1396
Illegal PstI site found at 1507 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1
Illegal PstI site found at 100
Illegal PstI site found at 1363
Illegal PstI site found at 1396
Illegal PstI site found at 1507
Illegal NgoMIV site found at 1449 - 1000COMPATIBLE WITH RFC[1000]
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