The average number of sequence reads that contained P. maxima diagnostic SNPs within this Olaparib other P. maxima database was 103 (± SE 9.15) and 62 (± SE 17.81) for the P. margaritifera SNPs within the P. margaritifera database. All putative biomineralisation genes (N = 7) were found to be expressed by the donor oyster (Fig. 2). Three of these genes N66, Perline and N44, were solely expressed by the donor with no expression from the host oyster. Here, the P. maxima diagnostic SNPs only detected expression of N66, Perline and N44 in the xenografts where P. maxima was the donor oyster

(Bs) and the P. margaritifera diagnostic SNPs only detected expression from the xenografts where P. margaritifera was the donor oyster (Sb) ( Fig. 2). For four of the seven biomineralisation genes (Linkine, Selleck BMS 907351 PfCHS1, MSI60 and Calreticulin), both donor and host oyster transcripts were detected within the xenografted pearl sacs (Bs, Sb; Fig. 2). Here, P. margaritifera SNPs detected expression of Linkine, PfCHS1, MSI60 and Calreticulin in the xenografts where P. margaritifera was the donor and host oyster (Sb, Bs) and P. maxima SNPs detected expression in the xenografts where P. maxima was the donor and host oyster (Bs, Sb), with the exception of Linkine ( Fig. 2). Gene transcripts from Calreticulin and MSI60, however, were detected in gonad

tissue samples from P. maxima and P. margaritifera. No specific amplification of Linkine and PfCHS1 transcripts was detected in the gonad samples. To further confirm the expression of biomineralisation genes from the host oyster and to validate the sequencing data (Illumina GAII), a highly informative region (40 bp in length) of Linkine was sequenced that contained five known species diagnostic single nucleotide polymorphisms (SNPs). Individuals from the allografted pearl sacs (Ss, N = 2; Bb, N = 2) were also sequenced to validate that the SNPs were species diagnostic, followed by sequencing of individuals from the xenografted pearl sacs (Sb, N = 5; Bs, N = 5) to determine whether the host or donor oyster species diagnostic SNPs were present ( Table 3). All P.

margaritifera allografted pearl sacs (Bb) showed an A nucleotide at a particular isothipendyl SNP site, whilst P. maxima allografted pearl sacs (Ss) had a T nucleotide at the SNP site. All five xenografted pearl sacs, where P. maxima was the donor oyster (Bs), had a P. maxima diagnostic SNP (T). Whilst four of the xenografts where P. margaritifera was the donor oyster (Sb) possessed the P. margaritifera SNP (A). However, one of these xenografted pearl sacs where P. margaritifera is the donor possessed the P. maxima diagnostic SNP (T), suggesting that the host was expressing Linkine in this individual ( Table 3). The other four diagnostic SNPs within the region sequenced for Linkine showed the same pattern as the above mentioned SNP site.