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采用RT-PCR、PCR-SSCP等分子生物学技术和关联分析等方法对虾夷马粪海胆Strongylocentrotusintermedius CYP51基因的cDNA序列进行了克隆和单核苷酸多态性分析。结果表明:虾夷马粪海胆的CYP51基因包括一个1 491 bp的开放阅读框,编码496个氨基酸;其开放阅读框的第161个核苷酸处存在1个单核苷酸多态性,即核苷酸发生了A→G突变。关联分析结果表明,该单核苷酸多态性与虾夷马粪海胆的体质量、性腺质量之间存在显著相关性(P<0.05)。本研究中首次克隆了虾夷马粪海胆CYP51基因的cDNA序列并进行了SNPs分析,研究结果可为虾夷马粪海胆的分子标记辅助育种提供参考。
The cDNA sequence of Strongylocentrotus intermedius CYP51 gene was analyzed by RT-PCR, PCR-SSCP and other molecular biology techniques and correlation analysis, and the single nucleotide polymorphism analysis was performed. The results showed that the CYP51 gene of horseradish shrimp (Eisenia fetipoensis) consisted of a 1 491 bp open reading frame encoding 496 amino acids and a single nucleotide polymorphism (SNP) at the 161st nucleotide of the open reading frame A → G mutations occur in nucleotides. Correlation analysis showed that there was a significant correlation between the single nucleotide polymorphism and the body weight and gonadal quality of sea urchin (E) (P <0.05). In this study, the cDNA sequence of CYP51 gene in sea urchin and shrimp (Eisenia abyonoidea) was cloned for the first time and SNPs were analyzed. The results of this study may provide reference for molecular marker-assisted breeding of Eriocheir japonica.