pone.0069647.g001

Pleurodeles waltl DNA polymerase ?.Schematic representation of P. waltl Pol? mRNAs (A). The short variant (Pol? ? Ex 8) presents a 103 bp deletion corresponding to exon 8. This deletion removes one part of the active site of the protein. The BRCT (breast cancer suppressor protein carboxy-terminal) and the Pol X domain, which contains the active site, are required for Pol? function. Pol? variants were amplified using primers indicated by horizontal arrows (B). Quantification of the obtained PCR products using a Gel Doc 2000 and the Quantity One v.4.3.1 software (Bio-Rad, Hercules, CA, USA) indicates that the short isoform represents 16% and 6% of the Pol? transcripts in the spleen and testis of P. waltl, respectively. Ctl??=?negative control. MW?=?100 bp DNA ladder. A cladogram was then constructed by neighbor joining (C) supported with 1000 bootstrap replications using the MEGA4 software (www.megasoftware.net/) and the following sequences: Terminal deoxynucleotidyl transferase (TdT); Bos taurus (DAA14763), Homo sapiens (BAB72001), Mus musculus (NP_001036693), Gallus gallus (NP_990720), Ambystoma mexicanum (AA092254), Xenopus laevis (NP_001079251), Danio rerio (AAS89780). Polymerase beta; Mus musculus (NP_035260), Homo sapiens (NP_002681), Xenopus laevis (NP_001081643), Xenopus tropicalis (AAH74537), Xiphophorus maculatus (AAU11319). Polymerase lambda; Danio rerio (NP_998408), Xenopus tropicalis (NP_001093716), Mus musculus (NP_064416), Homo sapiens (NP_001167555). Polymerase mu; Homo sapiens (NP_037416), Mus musculus (NP_059097), Danio rerio (NP_956542), Xenopus tropicalis (NP_001164987), Pleurodeles waltl (HE583591). The scale bar corresponds to the evolutionary distance.

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