Difference between revisions of "Part:BBa K1350007"

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===Usage and Biology===
 
===Usage and Biology===
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The expression levels of cel5A is repressed by glucose and induced by cellulose.It can hydrolyze β-1,4glucosan randomly and act on long cellulose chains. The main product is oligomeric glucose. It can digest glucosan to produce glucose monomer in high efficiency with synergy with β-1,4glucosidase.It shows high ability to hydrolyze the crystal cellulose but relative lower enzyme activity towards carboxymethyl cellulose, barley β-glucan, and PASC.
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Different from BBa_K805011 given by iGEM12_HUST-China, we have done the mutation of an ? to ?? and change the sequence in a way.
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However,because of the degeneracy of codon,the different sequence can encode the same enzyme.
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We construct the gene on the carrier displaying on the surface of Pichia pastoris and express it it in Pichia pastoris with electroporation. Then we detect the expression on the surface of Pichia pastoris by observing with fluorescent microscope and by flow cytometry analysis. 
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[[Image:Luorescent_microscope_3.jpg‎]]
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We fused FLAG-tag with N-terminal of cel5A gene on cel5A-pPIC9k vector. Label cells with Rabbit Anti Flag-Tag Polyclonal antibody and Goat Anti Rabbit RPE Polyclonal antibody. Result of flow cytometry analysis is as follows: fluorescence intensity x-mean value of Pichia pastoris with pPIC9k empty vector is 1.14, with cel5A-pPIC9k is 5.55.
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[[Image:Hust_part3.jpg]]
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[[Image:Hust_part4.jpg]]
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Left: pPIC9k negative control (x-mean: 1.14/13.4); Right: cel5A (x-mean: 5.55/16.5)
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1350007 SequenceAndFeatures</partinfo>
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Revision as of 15:54, 13 October 2014

Cel5A, endoglucanase（BBa_K805011 Improved）

we got this endoglucanase from iGEM kit,and we mutate it in a certain spot.[1]

Cel5A, endoglucanases

The endoglucanase genes cel5A can hydrolyze β-1,4glucosan randomly and act on long cellulose chains. The main product is oligomeric glucose. It can digest glucosan to produce glucose monomer in high efficiency with synergy with β-1,4glucosidase.It shows high ability to hydrolyze the crystal cellulose but relative lower enzyme activity towards carboxymethyl cellulose, barley β-glucan, and PASC.

Usage and Biology

Different from BBa_K805011 given by iGEM12_HUST-China, we have done the mutation of an ? to ?? and change the sequence in a way. However,because of the degeneracy of codon,the different sequence can encode the same enzyme.