Difference between revisions of "Team:Wageningen UR/Notebook/Chitinases"

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<h1><b>April</b></h1>
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<h1><b>July</b></h1>
 
<h2><b>Week 1</b></h2>
 
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<p>This week, we received our IDT order; this contained chitinase A and B from <i>Serratia marcescens</i> GEI strain, a strain which has been reported to cause higher mortality in <i<Varroa destructor</i> than in worker bees<sup><a href="#ch1" id="refch1">18</a></sup>. The chitinases were optimized for BioBrick use, as is described in their respective BioBrick pages: <a href="http://parts.igem.org/Part:BBa_K1913000"><i>chiA</i></a> and <a href="http://parts.igem.org/Part:BBa_K1913001"><i>chiB</i></a>.Initially, I tried to do a PCR to amplify the synthetic genes, but this resulted in a too small product for <i>chiB</i> (Figure 1).</p>
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<figcaption>Figure 1. Photo of the first plate with colonies (no. 15). These mites were not sterilized, so this was probably contamination.</figcaption>
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<figcaption>Figure 7. Cellular debris of <i>E. coli</i> BL21, the failed Cry1 and Cry2 clones, <i>Bacillus subtilis</i>, Bt HD350 and Bt tenebrionis, as well as isolates V46 and V47. The red and blue arrrow indicate bands which are probably Cry1Aa (133 kDa) and Cry3Aa (73 kDa). The ladder is a BioRad Precision Plus Protein Standard, the gel is a MiniProtean TGX 12%.</figcaption>
 
<figcaption>Figure 7. Cellular debris of <i>E. coli</i> BL21, the failed Cry1 and Cry2 clones, <i>Bacillus subtilis</i>, Bt HD350 and Bt tenebrionis, as well as isolates V46 and V47. The red and blue arrrow indicate bands which are probably Cry1Aa (133 kDa) and Cry3Aa (73 kDa). The ladder is a BioRad Precision Plus Protein Standard, the gel is a MiniProtean TGX 12%.</figcaption>
 
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<a id="ch1" href=http://www.sciencedirect.com/science/article/pii/S0022201110000522>1.</a> Tu, S., Qiu, X., Cao, L., Han, R., Zhang, Y., & Liu, X. (2010). Expression and characterization of the chitinases from Serratia marcescens GEI strain for the control of Varroa destructor, a honey bee parasite. Journal of invertebrate pathology, 104(2), 75-82.
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Revision as of 08:17, 11 October 2016

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Wageningen UR iGEM 2016

 

 

 

Main Results

July

Week 1

This week, we received our IDT order; this contained chitinase A and B from Serratia marcescens GEI strain, a strain which has been reported to cause higher mortality in than in worker bees18. The chitinases were optimized for BioBrick use, as is described in their respective BioBrick pages: chiA and chiB.Initially, I tried to do a PCR to amplify the synthetic genes, but this resulted in a too small product for chiB (Figure 1).

Figure 1. Photo of the first plate with colonies (no. 15). These mites were not sterilized, so this was probably contamination.

Week 2

Figure 1. Photo of the first plate with colonies (no. 15). These mites were not sterilized, so this was probably contamination.

Week 3

Week 4

May

Week 5

Week 6

Figure 5. Protein extracts of Bt HD350, E. coli with Cry1 and Cry2 and isolates V46, V47. The red arrow indicates a band that is probably Cry1. The ladder is a BioRad Precision Plus Protein Standard, the gel is a MiniProtean TGX 12%.

Week 7

Week 8

Figure 7. Cellular debris of E. coli BL21, the failed Cry1 and Cry2 clones, Bacillus subtilis, Bt HD350 and Bt tenebrionis, as well as isolates V46 and V47. The red and blue arrrow indicate bands which are probably Cry1Aa (133 kDa) and Cry3Aa (73 kDa). The ladder is a BioRad Precision Plus Protein Standard, the gel is a MiniProtean TGX 12%.

References

      1. Tu, S., Qiu, X., Cao, L., Han, R., Zhang, Y., & Liu, X. (2010). Expression and characterization of the chitinases from Serratia marcescens GEI strain for the control of Varroa destructor, a honey bee parasite. Journal of invertebrate pathology, 104(2), 75-82. ↩