Ementary Figure 1). All five putative transcripts contained a single open reading frame; however, only the kcnj2-12 transcript was the same length as the human KCNJ2 gene transcript (Supplementary Table 3). The structure of the KCNJ2 protein consists of two transmembrane domains (M1 and M2) linked by a pore region. The amino- and carboxyl-termini compose the cytoplasmic domain of the ion channel a
Rboxylterminus. The amplicon was subsequently introduced into the pGEM T-Easy vector (Promega) according to the manufacturer's instructions, and the products were bidirectionally sequenced to confirm their identity. The QuikChange SiteDirected Mutagenesis Kit (Stratagene) protocol was used to perform in vitro site-directed mutagenesis to create the 95?98 kcnj2 construct. The primers used in this p
Rminal repeats are typically inflexible, though alternative repeat -1 modules have recently been described (24,25). We tested acBat1 deletion derivatives to test if this paradigm applies to Bat1. First, we tested variants of acBat1 lacking 2 ( 18?0), 4 ( 16?0), 6 ( 14?0) or 8 ( 12?0) core repeats (Figure 5A and Supplementary Figure S10). The later half of repeat 20 and repeat +1 were retained in e
Ats are near identical and repeat order does not change the interface between repeats. Given the numerous nonRVD polymorphisms between Bat1 repeats, deletion or insertion of core repeats will always create novel repeat interfaces and should be experimentally validated before use in downstream applications. We next tested acBat1 derivatives where the 82 residues N-terminal of core repeat 1 (acBat1
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