草兔卵透明带3(ZP3)基因的克隆及序列分析

根据GenBank中欧洲兔ZP3 mRNA序列设计特异性引物, 以草兔卵巢组织总RNA为模板, 通过RT-PCR技术首次克隆得到草兔ZP3 cDNA编码区,命名为hZP3( GenBank登录号:KC447289)。生物信息学分析表明:扩增出的hZP3基因编码序列长1 260 bp, 编码419个氨基酸,包含一个完整的开放阅读框(ORF)。与欧洲兔ZP3基因核苷酸一致性高达96%。预测的ZP3蛋白具有ZP家族典型特征。系统进化树显示其与欧洲兔亲缘关系最近,其次是啮齿目动物,符合物种进化规律。hZP3基因编码区的成功克隆, 为进一步利用该基因通过免疫不育手段防治森林草兔危害提供了基础。

Cloning and Sequence Analysis of Lepus capensis Zona Pellucida 3 cDNA

In this study, specific primers were designed according to the ZP3 mRNA sequence of European rabbit (Oryctolagus cuniculus)in GenBank. Using total RNA isolated from hare(Lepus capensis)ovaries, the complementary deoxyribonucleic acid(cDNA)encoding hare ZP3, named hZP3, was amplified by reverse transcribed polymerase chain reaction(RT-PCR)for the first time(Accession number:KC447289). The hZP3 cDNA contains an open reading frame of 1 260 nucleotides encoding a polypeptide of 419 amino acid residues. Nucleotide identity between Lepus capensis and Oryctolagus cuniculus was 96%. The predicted ZP3 amino acids had the typical characteristics of ZP family. Phylogenetic tree analysis suggested that Lepus capensis had the closest phylogenetic relationship with the Oryctolagus cuniculus, followed by rodents, in line with the laws of the evolution of species. The successful cloning of hZP3 cDNA provided a theoretical basis for future immune-contraceptive study on hare control.