Identification of SS-like Peptide Precursors

Two transcripts encoding precursors of SS-like peptides were identified by analysis of A. rubens neural transcriptome sequence data—ArSSP1 (GenBank: KT601708) and ArSSP2 (GenBank: MN257487). The sequence of ArSSP1 has been reported previously [48 (link)]. The sequence of ArSSP2 is reported here for the first time and was identified on account of its similarity to precursors of SS-like peptides that have been identified in other echinoderms [49 (link)]. Informed by the assembled transcript sequences of ArSSP1 and ArSSP2, cDNAs comprising the complete open reading frame of ArSSP1 and ArSSP2 were amplified from A. rubens radial nerve cord cDNA by PCR using specific primers (electronic supplementary material, table S1) and Q5 High-fidelity DNA polymerase (NEB, Hitchin, UK; cat. no. M0491S), cloned into pBluescript II SK+ vector (Agilent Technologies, Santa Clara, USA; cat. no. 212205) and then sequenced (TubeSeq service; Eurofins Genomics, Ebersberg, Germany).

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Zhang Y., Yañez Guerra L.A., Egertová M., Zampronio C.G., Jones A.M, & Elphick M.R. (2020). Molecular and functional characterization of somatostatin-type signalling in a deuterostome invertebrate. Open Biology, 10(9), 200172.

Publication 2020

pBluescript II SK+ vector TubeSeq service

Cdna Cord Dna polymerase Echinoderms Neural Peptides Primers Radial nerve Rubens Transcriptome Vector

Corresponding Organization : Queen Mary University of London

Other organizations : University of Warwick

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Variable analysis

independent variables

Amplification of cDNAs comprising the complete open reading frame of ArSSP1 and ArSSP2 from A. rubens radial nerve cord cDNA by PCR using specific primers

dependent variables

Sequences of ArSSP1 and ArSSP2

control variables

Use of Q5 High-fidelity DNA polymerase (NEB, Hitchin, UK; cat. no. M0491S) for PCR amplification
Cloning of amplified cDNAs into pBluescript II SK+ vector (Agilent Technologies, Santa Clara, USA; cat. no. 212205)
Sequencing of cloned cDNAs (TubeSeq service; Eurofins Genomics, Ebersberg, Germany)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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