sanger sequencing service  (Azenta)

Journal: The Journal of Cell Biology

Article Title: Insights into retinal disease and non-tubulin glutamylation from a RPGR–TTLL5 complex structure

doi: 10.1083/jcb.202508020

Figure Lengend Snippet: A minimal C-terminal helix in the RPGR BD is sufficient for CID binding and intercalates into the CID helical bundle through a conserved aromatic network. (A) Domain organization of human TTLL5 and RPGR. TTL catalytic core, blue; cMTBD, maroon; CID, green. RCC1-like β-propeller domain in RPGR, yellow, with each repeat shown as a separate propeller blade. Sequence for the minimal BD peptide required for TTLL5 targeting to RPGR shown in magenta at the top. (B) ITC measurements of WT miniBD peptide titrated into CID. Best fit of n = 2 independent experimental replicates. (C) 2.8-Å X-ray crystal structure of human TTLL5 CID in complex with human RPGR miniBD shown in cartoon representation; CID, green; RPGR, magenta. (D and E) Key conserved hydrophobic interactions at the CID–BD interface. Solid black boxes around residue labels designate disease-associated mutations, dashed boxes designate mutations without disease annotation in the ClinVar database that we predict are pathogenic based on our structure. RPGR and TTLL5 are colored as in B; H-bonds are shown as dashed lines.

Article Snippet: Vectors were sequenced by Psomagen standard Sanger sequencing service.

Techniques: Binding Assay, Sequencing, Residue

Journal: The Journal of Cell Biology

Article Title: Insights into retinal disease and non-tubulin glutamylation from a RPGR–TTLL5 complex structure

doi: 10.1083/jcb.202508020

Figure Lengend Snippet: Multiple sequence alignment of vertebrate TTLL5 CID and RPGR BD. (A) TTLL5 CID. (B) RPGR miniBD. Secondary structure elements indicated above the sequence and based on our X-ray crystal structure of the complex; gaps of disordered, unmodeled residues are indicated by a dotted line. Hydrophobic residues at the CID–BD interface indicated with blue boxes. Residues mutated for ITC experiments indicated with black arrows above the sequence.

Article Snippet: Vectors were sequenced by Psomagen standard Sanger sequencing service.

Techniques: Sequencing

Journal: Biofilm

Article Title: Experimental evolution in the cystic fibrosis chemical environment reveals early TCA cycle flux as a central regulator of Mycobacterium abscessus biofilm formation

doi: 10.1016/j.bioflm.2025.100343

Figure Lengend Snippet: Experimental evolution of MAB 0253a in SCFM1 selects for novel biofilm phenotypes. A) Graphical overview of the evolution experiment. SNG stands for S CFM1 n o g lucose B) Colony morphologies of all evolved populations from passage 15 of the evolution experiment on TYEM agar plates. The smooth variant MAB 0253a (ancestor) and the rough variant MAB 0253b are included for comparison. C) Aggregation dynamics of all evolved populations from passage 15 of the evolution experiment in TYEM media. The left plot depicts the peak aggregate OD 600 measurement of each lineage, taken within 40–52 h post-inoculation. The right plot depicts the aggregate OD 600 readout taken at 68 h post-inoculation. P-value obtained by unpaired Student's T-test against the Ancestor. ∗ denotes P-value <0.05. D) Graphical overview of MAB_0812 ( mraA ) and MAB_0813c ( mraB ) on the positive and negative strands, with nucleotide positions on the contiguous genome sequence labelled on the horizontal axis. Mutations observed in laboratory evolution experiments are denoted with a blue point. The helix-turn-helix (HTH) DNA-binding domain is green and the C-terminal effector-binding domain is red. Allelic variants for orthologs isolated from people with cystic fibrosis are denoted with red points and are labelled with their amino acid positions on each gene. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: PCR products with the correct fragment size were purified and sequenced using the Sanger Sequencing Services at Azenta Life Sciences.

Techniques: Variant Assay, Comparison, Sequencing, Binding Assay, Isolation