Difference between revisions of "Part:BBa K2938006"

Latest revision as of 09:34, 12 October 2019

Cry4Ba + HA tag

Cry4Ba Bti toxin with HA-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 Cry4Ba is a 128 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)[1], 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.

Cry4Ba is toxic specifically against Aedes Aegypti mosquitoes’ larvae (compared to Cry4Aa) [2]

Amplification

PCR amplification of Cry4Ba by PCR.

Usage and Biology

The Cry4Ba is a toxin with larvicidal effect on mosquitos’ larvae, and HA-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 XbaI site found at 1708
Illegal SpeI site found at 2397
Illegal PstI site found at 212
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 96
Illegal SpeI site found at 2397
Illegal PstI site found at 212
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 1708
Illegal SpeI site found at 2397
Illegal PstI site found at 212
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 1708
Illegal SpeI site found at 2397
Illegal PstI site found at 212
Illegal NgoMIV site found at 2520
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 502

@@ Line 5: / Line 5: @@
 Cry4Ba Bti toxin with HA-tag attached in the N- terminal in order to be able to detect the translated protein in western blot (using antibodies against the tag)
-<!-- Add more about the biology of this part here
+===Description===
-===Usage and Biology===
+Pesticidal crystal protein Cry4Ba is a 128 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)[1], 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]]
+Cry4Ba is toxic specifically against Aedes Aegypti mosquitoes’ larvae (compared to Cry4Aa) [2]
+===Amplification===
+[[File:Cry4Ba PCR.jpeg|200px|thumb|left|PCR  amplification of Cry4Ba by PCR.]]
+<!-- Add more about the biology of this part here -->
+===Usage and Biology===
+The Cry4Ba is a toxin with larvicidal effect on mosquitos’ larvae, and HA-tag is used to detect proteins in western blot using antibodies specific to the tag.
 <!-- -->
 <span class='h3bb'>Sequence and Features</span>