【摘要】 采用cDNA末端快速扩增技术(rapid amplification of cDNAends,RACE)获得了1 672 bp的斑节对虾钙网蛋白(Penaeusmonodon calreticulin,PmCRT)基因cDNA序列。该序列包含37 bp的5’非编码区(UTR)和414 bp的3’非编码区(UTR)以及1 221 bp的开放阅读框(ORF),可编码406个氨基酸。预测的分子量约为46.8ku,理论等电点为4.13。序列比对分析表明PmCRT与中国明对虾(Fenneropenaeus chinensis)的相似性和同源性最高,分别为100%和98.8%。实时荧光定量PCR对组织表达检测的结果可知,PmCRT的mRNA在卵巢、肝胰腺、肌肉、脑、心脏、胃、眼柄和肠均有表达,卵巢的表达量最高,肌肉次之;对卵巢发育6期变化检测可知,PmCRT的mRNA在Ⅲ期卵巢表达量最高,Ⅰ期的表达量最低。以期为进一步研究该基因在斑节对虾卵巢发育中的作用提供基础数据。研究亮点:目前对斑节对虾卵巢发育的分子调控机理还知之甚少,有大量卵巢发育相关基因还不为人所知。本文研究了钙网蛋白在斑节对虾卵巢发育各期的表达谱,为阐明斑节对虾卵巢发育分子调控机理提供基础数据。更多还原

【Abstract】 The techniques of Rapid Amplification of cDNA Ends were used to clone the Penaeus monodon calreticulin gene(PmCRT).The full length cDNA of PmCRT contained a 5′untranslated region(UTR) of 37 bp,an ORF of 1 221 bp encoding a polypeptide of 406 amino acids with an estimated molecular mass of 46.8 ku and a 3′UTR of 414 bp.Sequence alignment analysis showed that the PmCRT and Fenneropenaeus chinensis shared a similarity of 100%,and a homology of 98.8%.The quantification result of PmCRT mRNA in 8 tissues of Penaeus monodon showed that,in ovary the highest levels,followed by muscle,intestines,stomach,eyestalk,hepatopancreas,heart and brain.Furthermore,the PmCRT expression was found to be at high level in the six ovarian stages of development and the expression of ovarian growth levels in the third growth phase was significantly higher than that of others,while the lowest in the first growth phase.The study provided essential materials and laid a firm foundation for the further research of the function of PmCRT in ovary development.更多还原