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Manual gene annotation was performed by finding missing start/stop codons, by defining correct exon/intron borders, by analyzing perfectly matching ESTs and by taking into account the amino acid conservation of the thaumatin domain (Pfam: PF00314) and in particular the position of conserved cysteines.
They are classified according to the relative position of conserved N-terminal cysteine residues, in which CC chemokines represent the most abundant family and have the first 2 cysteines placed adjacently [26], [27].
Visual inspection of the conserved sequences among the human, chimpanzee, mouse, rat, and chicken genomes (UCSC genome browser hg16 build, table mxPt1 Mm3RnGg_pHMM) along the regions covered by our array suggested a correlation of origin peaks with the position of conserved elements.
The most obvious result in our study relates the variation in the position of conserved motif IV with a structural explanation for the poor base specificity and promiscuous nucleotide hydrolysis reported in nucleotide binding studies of FtsY by Shan and Walter [24].
Chemokines are a family of chemoattractive cytokines and are grouped into C, CC, CXC, and CX3C subfamilies based on the position of conserved cysteine residues [ 3, 4].
This observation was supported with a genome-wide analysis whereby the relative position of conserved and divergent CTCF sites was determined with respect to all domains in the mouse genome.
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Three dimensional modeling of the protein sequence showed that the mature Da-CTSL1 protein folds into an expected cathepsin L structure producing a substrate binding pocket with appropriate positioning of conserved amino acid residues.
It appears that T/A→G/C substitutions at 5' flanking positions of conserved CpG sites are significantly more frequent in fish genes compared to sea squirt genes.
We took the α of Cs genes as a reference for those of the fish genes and assumed that the two flanking positions of conserved CpG sites have equal opportunities in obtaining C or G (Figure 1).
In the test, the nucleotide frequencies at the TpG flanking positions of conserved NNTGNN dicodons (the dicodon presents in both homologous fragments) were obtained using the homologous fragments between Ci and Cs.
The model quality for these domains was superior to that for the IR domains and the models adopted a similar fold to characterised TK domains, including the positioning of conserved residues important for function, such as Tyr1189 and 1190 in HIR, which are the most striking features of the 1IRK structure and are autophosphorylated upon insulin binding the active site of the enzyme.
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