波斯锐缘蜱HSPC1和HSPC4基因的全长克隆及生物信息学和差异表达分析OA

This study aims to obtain the full-length sequences of the Argas persicus HSPC1(ApHSPC1)and HSPC4(ApHSPC4)genes and analyze the bioinformatic features and differential expression levels of the two genes.The rapid amplification of cDNA ends(RACE)was used to amplify the missing sequences of the 3′and 5′ends of ApHSPC1 and ApHSPC4.The full-length cDNA sequences of ApHSPC1 and ApHSPC4 were deter-mined by sequencing and assembly,and the biological information of the two gene-encoded proteins was analyzed by bioinformatics tools.Phylogenetic trees were constructed to examine evolutionary rela-tionships.qPCR was used to measure the relative expression levels of ApHSPC1 and ApHSPC4 in different developmental stages and tissues of A.persicus.The results showed that the full-length cDNA sequences of ApHSPC1 and ApHSPC4 were determined to be 2 430 bp and 2 637 bp,respectively,with open reading frames of 2 178 bp and 2 370 bp.Both encoded proteins lacked signal peptides and contained distinct localization motifs,namely the cytosolic MEEVD motif in ApHSPC1and the endoplasmic reticulum retention HDEL motif in ApHSPC4.Sequence similarity and phylogenetic analyses showed that both genes exhibited high homology and a close evolutionary relationship with heat shock protein 90(HSP90)gene from Ornithodoros turicata.qPCR analysis revealed that ApHSPC1 expression was consistently higher than ApHSPC4 across all develop-mental stages(P＜0.001).Under unfed conditions,ApHSPC1 expression in the ovary,midgut,and salivary glands was significantly higher than that of ApHSPC4(P＜0.001).After blood feeding,ApHSPC1 remained more highly expressed in the ovary and midgut(P＜0.001),whereas no significant difference was observed between the two genes in the salivary glands(P＞0.05).Additionally,ApHSPC1 showed significantly higher expression in the ovary than in the testis(P＜0.001),while ApHSPC4 displayed the opposite trend(P＜0.001).These findings indicate that ApHSPC1 and ApHSPC4 possess distinct protein localization signals and exhibit differential expression patterns across developmental stages and tissues,suggesting that they may play divergent roles in development,blood meal digestion,and reproduction in A.persicus.