Ank3-exon1e Mouse Luciferase Reporter (Royal College of Surgeons in Ireland)
Royal College of Surgeons in Ireland
12 St Stephens Green
Resource:mouse ank3-promoter firefly luciferase reporter gene. The following primers were used to PCR amplify mouse ank3 gene specific reporter-gene constructs from mouse total DNA preparations. 5’-GCTAGCTAGCCTTTCCTTGAGATCAGGGGAAGT-3’ and 5’-TTAGGTACCGGGTGGGGCAACCGC-3’, with a PCR product size of 1903 bp, which were cloned into pGL3-Basic vector (Promega) using the restriction enzymes, KpnI and NheI (Fermentas, Life Technologies, Dun Laoghaire, Ireland). The amplified sequence is located directly upstream and contained in the region encoding exon 1e 5, expressed in ankyrin-G transcripts of approximately 93 kDa, 192 kDa, 209-214 kDa and also 500 kDa. Primers for obtaining the first and second half segments of the promoter were used in conjunction with the above primers to obtain these promoter constructs as follows, 5’- GCTAGCTAGCCTGGCTTTTTCTGTGCCAGAA-3’ and 5’- TTAGGTACCTTCTGGCACAGAAAAAGCCAG-3’. Amplified sequences were verified by sequencing using RV3 standard sequencing primers (MWG, Germany). For preparation of the ankyrin-3 mRNA directed constructs we used the previously published sequence ‘CT19’ -5’-GAGACATAAACTGGCCAAC-3’ 2 cloned into pSilencer2.1.
Neurodegenerative disease in general
Genetic Material (e.g. DNA, RNA, vectors)
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