克氏原螯虾核糖体蛋白S24(RPS24)基因的克隆与表达分析

Cloning and expression analyses of ribosomal protein S24 (RPS24) gene in Procambarus clarkii

为了阐明克氏原螯虾(Proambaus clarkii)核糖体蛋白S24(ribosomal protein S24,RPS24)基因序列特征及其在卵巢发育中的作用,本研究利用RACE方法克隆获得了克氏原螯虾RPS24(PcRPS24)基因全长cDNA序列,并利用实时荧光定量PCR(QPCR)方法对该基因的表达模式进行了分析。结果显示:PcRPS24 cDNA序列全长438 bp,编码136个氨基酸,并具有11个磷酸化位点和1个RPS24e标志序列;克氏原螯虾与八棘多刺鱼的RPS24氨基酸序列相似性最高,与昆虫纲动物在进化上的亲缘关系最近;PcRPS24基因表达量在克氏原螯虾发育过程中逐渐升高,在卵巢发育至Ⅰ期的成虾中达到最大值;在成虾的不同组织中,PcRPS24基因在肝胰腺和卵巢中的表达量较高;在Ⅰ-Ⅵ期的卵巢中,PcRPS24基因在Ⅰ期卵巢中的表达量最高。结果表明PcRPS24可能在克氏原螯虾早期卵巢发育中起重要作用。

In order to explore the sequence characteristics and the function in ovarian development of ribosomal protein S24(RPS24) gene in Procambarus clarkii(PcRPS24 gene),the PcRPS24 cDNA sequence was obtained by the rapid amplification of cDNA ends(RACE) and the expression pattern of this gene was determined by quantitative real-time PCR(QPCR).The results showed that the full-length cDNA of PcRPS24 was 438 bp, encoding a deduced protein with 136 amino acids;11 phosphorylation sites and a RPS24 e signature were predicted in the amino acid sequence of PcRPS24;RPS24 amino acid sequence of P.clarkii and Pungitius pungitius were the most consistent, and PcRPS24 was more closely related to RPS24 from invertebrate insects in evolution;The expression level of PcRPS24 gene gradually increased during the developmental period of P.clarkii,and reached a maximum value in adult P.clarkii with stage I ovary.In different tissues of adult P.clarkii,PcRPS24 gene was highly expressed in hepatopancreas and ovary;In the Ⅰ-Ⅵ stage of ovaries in P.clarkii,PcRPS24 gene expression level was the highest in stage I ovary.These results indicate that PcRPS24 may play an important role in the early stage of ovarian development in P.clarkii.