Difference between revisions of "Part:BBa K2938005"
 
Line 4: Line 4:
  
 
Cry11Aa Bti toxin with Strep-tag attached in the N- terminal in order to be able to detect the translated protein in western blot (using antibodies against the tag)
 
Cry11Aa Bti toxin with Strep-tag attached in the N- terminal in order to be able to detect the translated protein in western blot (using antibodies against the tag)
 +
 +
===Description===
 +
 +
Pesticidal crystal protein Cry11Aa is a 72 kDa protein which can be isolated from Bacillus thuringiensis subsp. israelensis (Bti).
 +
The pBtoxis is a toxin-encoding plasmid that its origin is the Bti and it encodes to four Cry toxins (Cry4Aa, Cry4Ba, Cry10Aa and Cry11Aa) and three Cyt toxins (Cyt1Aa, Cyt2Ba and Cyt1Ca), which are specifically toxic against Aedes, Anopheles and Culex mosquitoes which are vectors to many diseases caused by parasites and viruses, such as Malaria, Zika and Dengue.
 +
Each of the toxins hold a low level of larvicidal effect but their combination has a synergetic effect which is stronger than the linear combination we expect. The Cyt1Aa toxin effect in the synergetic effect is mainly the prevention of resistance in the mosquito to the Cry toxins.
 +
The bigger Cry toxins, Cry4Aa and Cry4Ba have a conserved C-terminal end which helps the toxin fold correctly and form crystal structures.
 +
 +
[[File:ToxinTable.png]]
 +
 +
Cry11Aa is toxic mostly against Aedes Aegypti mosquitoes’ larvae
 +
 +
 +
===Amplification===
 +
[[File:Cry11Aa PCR.jpeg|200px|thumb|left|PCR  amplification of Cry11Aa by PCR.]]
 +
  
 
<!-- Add more about the biology of this part here -->
 
<!-- Add more about the biology of this part here -->

Latest revision as of 09:32, 12 October 2019


Cry11Aa + Strep tag

Cry11Aa Bti toxin with Strep-tag attached in the N- terminal in order to be able to detect the translated protein in western blot (using antibodies against the tag)

Description

Pesticidal crystal protein Cry11Aa is a 72 kDa protein which can be isolated from Bacillus thuringiensis subsp. israelensis (Bti). The pBtoxis is a toxin-encoding plasmid that its origin is the Bti and it encodes to four Cry toxins (Cry4Aa, Cry4Ba, Cry10Aa and Cry11Aa) and three Cyt toxins (Cyt1Aa, Cyt2Ba and Cyt1Ca), which are specifically toxic against Aedes, Anopheles and Culex mosquitoes which are vectors to many diseases caused by parasites and viruses, such as Malaria, Zika and Dengue. Each of the toxins hold a low level of larvicidal effect but their combination has a synergetic effect which is stronger than the linear combination we expect. The Cyt1Aa toxin effect in the synergetic effect is mainly the prevention of resistance in the mosquito to the Cry toxins. The bigger Cry toxins, Cry4Aa and Cry4Ba have a conserved C-terminal end which helps the toxin fold correctly and form crystal structures.

ToxinTable.png

Cry11Aa is toxic mostly against Aedes Aegypti mosquitoes’ larvae


Amplification

PCR amplification of Cry11Aa by PCR.


Usage and Biology

The Cry11Aa is a toxin with larvicidal effect on mosquitos’ larvae, and strep-tag is used to detect proteins in western blot using antibodies specific to the tag.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1719
    Illegal SpeI site found at 831
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1719
    Illegal SpeI site found at 831
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1719
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1719
    Illegal SpeI site found at 831
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1719
    Illegal SpeI site found at 831
  • 1000
    COMPATIBLE WITH RFC[1000]