Difference between revisions of "Part:BBa K1955003"
 
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<b style="font-size:23px;">pSB1C3-5'HYG</b><br><br>
 
<b style="font-size:23px;">pSB1C3-5'HYG</b><br><br>
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The promoter and the ribosomal binding site of Leishmania genome has not been elucidated yet. We selected the 5'- untranslated region of a highly expressed gene, P36, to be the promoter, RBS binding site and other extra function needed for Leishmania protein expression. We also added a Hygromycin resistant gene as drug selection marker. AS a dual functional biobrick of regulatory and selection marker, we provide the user the regulation of protein expression and also the drug selection system that is most commonly and effectively used in Leishmania experiments.
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We replaced p36 and nagt genes with a cloning site for an exogenous gene and with hyg as a selection marker, respectively. This dual-function biobrick enables stable expression of foreign proteins by drug selection in Leishmania.<br><br>
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This biobrick needs to use with the 3'UTR that we provide as the terminator of the Leishmania to perform a complete protein expression system for Leishamnia. Add the protein sequence in between the Leish-5'UTR-HYG and Leish-3'UTR for leishmania to expression your target protein.<br><br>
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<b style="font-size:20px;">(1) The basic part checked by PCR: </b>
 
<b style="font-size:20px;">(1) The basic part checked by PCR: </b>
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Latest revision as of 08:12, 5 December 2016

pSB1C3-5'HYG

We replaced p36 and nagt genes with a cloning site for an exogenous gene and with hyg as a selection marker, respectively. This dual-function biobrick enables stable expression of foreign proteins by drug selection in Leishmania.

(1) The basic part checked by PCR:

We used pSB1C3-5’HYG, pSB1C3-3’UTR, pSB1C3-HA, pSB1C3-OVA as template, to check the length of the inserts. The PCR reaction was performed with Taq polymerase, and screened in 0.8％ agarose gel by electrophoresis.





Sequence and Features BBa_K1955003 SequenceAndFeatures