Difference between revisions of "Part:BBa K1217003"

(→‎Expression of ppk1 enzyme)
(→‎Usage and Biology)
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Efficiency of this construct is compared with Two other designs  [https://parts.igem.org/Part:BBa_K1217008 BBa_K1217008] and [https://parts.igem.org/Part:BBa_K1217010 BBa_K1217010].
 
Efficiency of this construct is compared with Two other designs  [https://parts.igem.org/Part:BBa_K1217008 BBa_K1217008] and [https://parts.igem.org/Part:BBa_K1217010 BBa_K1217010].
 
We test the constructs' efficiency from 2 parameters: poly-P synthesis efficiency and phosphate removal efficiency from the medium.
 
We test the constructs' efficiency from 2 parameters: poly-P synthesis efficiency and phosphate removal efficiency from the medium.
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[[File:Ppkonly.png|150px|thumb|none|Expected model of BBa_K1217003 ]]
  
 
==='''Expression of ppk1 enzyme'''===
 
==='''Expression of ppk1 enzyme'''===
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[[File:Ppkonly.png|150px|thumb|none|Expected model of BBa_K1217003 ]][[File:HKU13_KOppkSDS.png|400px|thumb|none|SDS-PAGE analysis of expression of K.oralis PPK1 enzyme inside E.coli. Expression of PPK1 is induced by 0.5mM IPTG at 30 degree for 5 hours. Band marked with star is the PPK1 enzyme, the enzyme expressed is soluble. ]]
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We confirm the expression of PPK1 enzyme in e.coli.
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[[File:HKU13_KOppkSDS.png|400px|thumb|none|SDS-PAGE analysis of expression of K.oralis PPK1 enzyme inside E.coli. Expression of PPK1 is induced by 0.5mM IPTG at 30 degree for 5 hours. Band marked with star is the PPK1 enzyme, the enzyme expressed is soluble. ]]
  
 
==='''Efficiency in Poly-P synthesis'''===
 
==='''Efficiency in Poly-P synthesis'''===

Revision as of 05:31, 5 October 2013

pT7-ppk1(K.oralis)

Polyphosphate kinase (ppk1) (E.C. 2.7.4.1) catalyzes the formation of polyphosphate from ATP. In BBa_K1217003, ppk1 was under the control of T7 promotor. Under iptg induction, over-expression of ppk1 can reduce the phosphate level in the medium by its incorporation into polyphosphate synthesis inside the bacteria.

Usage and Biology

Efficiency of this construct is compared with Two other designs BBa_K1217008 and BBa_K1217010. We test the constructs' efficiency from 2 parameters: poly-P synthesis efficiency and phosphate removal efficiency from the medium.

Expected model of BBa_K1217003

Expression of ppk1 enzyme

We confirm the expression of PPK1 enzyme in e.coli.

SDS-PAGE analysis of expression of K.oralis PPK1 enzyme inside E.coli. Expression of PPK1 is induced by 0.5mM IPTG at 30 degree for 5 hours. Band marked with star is the PPK1 enzyme, the enzyme expressed is soluble.

Efficiency in Poly-P synthesis

Efficiency in phosphate removal from the environment

Testing

Sequence and Features


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 309