case_images: PMC5996431_01
case_images: Schematic representation of in silico analysis of the c.2678-15C>A variant identified in intron 22 of the FBN1 gene by various computational tools. (a) Representation of the position of the canonical (c.2678, black triangle) and cryptic (c.2678-13, grey triangle) splice site identified in the reference (RS) and mutated sequence (MS), respectively. Arrow: the C to A point mutation (c.2678-15C>A). The intron 22 (i22) sequence is shown in lower case and exon 23 (E23) sequence in upper case. (b) The predicted splice score of the canonical site decreases by 29.8% and 49.7% in the mutated sequence (grey parallelogram) versus the reference sequence (black parallelogram) as revealed by the MES and GSP algorithm, respectively. (c) The predicted splice scores of a new cryptic splice site due to the c.2678-15C>A mutation.
case_images: BMRI2018-3536495.001.jpg
case_images: PMC5996431_01_BMRI2018-3536495.001.jpg
case_images: fig1
case_images: PCR fragments were cloned into a linearized dual tagged (N-terminal 3xFLAG and C-terminal c-Myc) expression vector (p3xFLAG-Myc-CMV-26, Sigma) as detailed in Supp.  Figure 1A. Plasmids grown in XL1-Blue Supercompetent E. coli cells (Stratagene) were purified by using a PureLink HiPure plasmid filter purification kit (Invitrogen) according to the manufacturer's protocol. Plasmid 1283 was selected as the FBN1-Ref minigene, and plasmid 1288 was selected as the FBN1-Mut minigene (Supp.  Figure 1B).