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In silico validation of this assembled transcript is achieved by translating the sequence (http://web.expasy.org/translate) and by searching protein databases using the Protein Basic Local Alignment Search Tool Standard Protein BLASTT, http://blast.ncbi.nlm.nih.gov; Figure S4B).
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We also evaluated the potential protein coding capacity of sRNAs by translating the sequences in all three open reading frames.
The coding regions were aligned in a codon-based manner, by translating the nucleotide sequences into peptide sequences with transeq included in the EMBOSS package and than aligning them with MUSCLE, version 3.6 [ 41].
Analysis was then performed by translating the DNA sequence to protein using the ExPASy translate tool (Swiss Institute of Bioinformatics; http://web.expasy.org/translate/).org/translate/
Putative protein sequences were obtained by translating the cDNA sequences using the EMBOSS program "Transeq" [ 57].
Comparisons were made at the DNA level using Nucmer and then the protein level using Promer by translating the DNA sequences in all six reading frames.
Putative introns were identified by translating the DNA sequences in the three frames and comparing the results with a manual alignment of known amylase proteins.
The most likely ORF of the Hpa set of sequences was identified by translating the assembled EST sequences and singletons in 3 positive frames and selecting the longest ORF.
The first step is to prepare protein sequence databases by translating the RefSeq DNA into all six frames: genome NC_004088, and plasmids NC_004836, NC_004837, NC_004838.
By translating the CDS to an amino acid sequence, we can make further analysis at the protein level, such as the statistics of amino acids, the identification of signal peptide, the prediction of potential TM regions, helical wheel display of these potential TM helices and generally gain an overview of the protein.
Bear protein sequences were obtained by translating the bear transcripts using the same open reading frame as in other species.
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