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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We selected the 5'- untranslated region of a highly expressed gene, P36, since the 5’ UTR serves as stabilizing the mRNA for constitutive expression in both Leishmania stages. 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 selected the 5'-untranslated region (UTR) of a highly expressed gene p36 because the 5’UTR stabilizes the mRNA for constitutive expression in both Leishmania stages. We also added a hygromycin-resistant gene as a drug selection marker. This dual-function biobrick enables the regulation of protein expression and drug selection. The 3'UTR serves as the terminator for protein expression in Leishamnia. This system is most commonly and effectively used in Leishmania experiments. Users can insert any protein sequence between the Leish-5'UTR-HYG and Leish-3'UTR for your target protein expression 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>

Revision as of 10:31, 4 December 2016

pSB1C3-5'HYG

We selected the 5'-untranslated region (UTR) of a highly expressed gene p36 because the 5’UTR stabilizes the mRNA for constitutive expression in both Leishmania stages. We also added a hygromycin-resistant gene as a drug selection marker. This dual-function biobrick enables the regulation of protein expression and drug selection. The 3'UTR serves as the terminator for protein expression in Leishamnia. This system is most commonly and effectively used in Leishmania experiments. Users can insert any protein sequence between the Leish-5'UTR-HYG and Leish-3'UTR for your target protein expression 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