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 (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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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>
  
 
<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>

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