Searching DNA sequence of artificial intron (ie, splice site study) for inserting in plasmid (for drosophila cell system)?

28 March 2019 1 9K Report

I want to construct a plasmid (for Drosophila cell system) containing a intron in order to study splicing process. In detail, I would like to insert wether a weak or a strong splice donor site (followed by small intron+ splice acceptor) in the firefly luciferase to be able to easy monitor the splicing activity by standard luciferase assay. However, if I can find easily the consensus site for 5'SS and 3'SS, I am struggling to find the appropriate full DNA sequences that I would like to practically insert in my plasmid. Is there someone that can help me finding this sequence? (addgene reference? detailled publication with the DNAsequence fully available ?)

Thanks in advance for your help,

Baptiste Panthu

Hi Julie,

Comment vas-tu?

J'ai un plasmide pcdna3.1 (hygro-) avec un intron dans la 5'UTR en amont de la renilla luciferase. L'épissage a bien lieu et semble charger l'ARNm en protéine régulatrice permettant une synthèse protéique forte lors d'une transaction en cellules. Si tu veux je t'envoie le plasmide ainsi que sa séquence annotée (Ape plasmid editor). Baptiste

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