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The raw data and normalized data are available under Gene Expression Omnibus, one of the largest MIAME compatible expression profile resources on the web, under accession GSE17846.
From a list of 8 housekeeping genes (HKG), we chose HMBS and ALDOA, which meet the criteria of less variation between samples and compatible expression level with the studied genes.
A series of four vectors were constructed by Horanyi et al. [25] in which the GATEWAY™ recombination cassette from Invitrogen was combined with the four compatible expression vectors of the Novagen (EMD Chemicals, San Diego, CA) DUET vector system (Table 1).
Once the single-copy status has been documented the respective candidate sites lend themselves to the FLP-mediated integration of compatible expression cassettes.
When, in addition, we look at compatible expression for different iterations (3 and 10) we still find over 50% in agreement.
The versatility of the Gateway system allows easy transfer of cDNAs into any compatible expression system for further interaction or functional studies.
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We also constructed a Gateway-compatible expression vector, pRetroTRexD30, on the MSCV back bone.
For the Tet-On Gateway-compatible expression vector, the original vector was used as a negative control.
An estrogen-inducible, gateway-compatible expression vector, pFZ19, was used to generate transgenic Arabidopsis plants (Zhang et al. 2010).
The clones from the entry vectors can be easily shuttled into different Gateway-compatible expression vectors of many types in a high-throughput fashion [ 5, 6].
Following sequence verification, cDNAs were transferred into the Gateway-compatible expression vectors TSHgwICPB or TgwSHICPB with N- RIPK3) or C-termiN- RIPK3L, MLKL S358D) Strep-HA tag, respectively.
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