草原兔尾鼠卵透明带3(ZP3)cDNA保守序列的克隆与序列分析  

Cloning and Sequencing Analysis of Lagurus lagurus Zona Pellucida 3 cDNA Conservative Fragment

作　　者：张富春[1] 张蕴斌[1] 李轶杰[1] 张渝疆[2] 李阔宇[1] 钟哲[1] 马正海[1] 恩特马克[1] 

机构地区：[1]新疆大学生命科学与技术学院分子生物学重点开放实验室,新疆乌鲁木齐830046 [2]新疆地方病防治研究所,新疆乌鲁木齐830002

出　　处：《地方病通报》2001年第4期70-73,共4页Endemic Diseases Bulletin

基　　金：中国科学院西部之光项目;科技部攻关项目 ( TJ99LA0 1 );教育部骨干教师项目资助

摘　　要：根据 Gene Bank数据库中 5种啮齿目动物卵透明带 3 (ZP3 ) c DNA的序列 ,利用 DNAMAN和 NIH NCBI BLAST同源性分析程序设计了特异性引物。从 6周龄的未孕雌性的草原兔尾鼠卵巢中用柱离心法得到总 RNA,利用反转录技术得到 c DNA,通过特异性引物进行 PCR扩增草原兔尾鼠的 ZP3 c DNA保守序列。将扩增产物连接到 p MD1 8- T载体上 ,通过蓝白斑筛选得到阳性克隆 ,酶切鉴定后进行测序。将其与 Gene Bank中啮齿目动物 ZP3 c DNA进行同源性比较 ,获得了草原兔尾鼠 ZP3 c DNA的保守基因序列 ,与啮齿类同类基因比较 ,具有较高的同源性。The sequences of five Rodent zona pellucida 3 (ZP3) cDNAs were obtained from the Gene Bank. According to the sequences, the special primers were designed by DNAMAN and NIH NCBI BLAST. The ovaries were prepared by dissecting the nonpregnant female Lagurus lagurus distributed in Xinjiang. The total RNA was abstracted from the ovaries and cDNA of Lagurus lagurus ZP3(lZP3) was cloned and ligated into pMD18 T Vector. The correction of the recombinant vector pMD18 lZP3 was tested by blue and white spot screen and restriction enzyme digestion. Finally, the nucleotide sequence of the cloned lZP3 cDNA was defined by PE 377 DNA Sequencer (Using RV M and reverse M13 47 as the primers). The results showed that the conservative sequence of the cloned lZP3 was high similar to five published Rodent ZP3 cDNAs.

关 键 词：草原兔尾鼠 卵透明带3 cDNA保守序列 序列分析 

分 类 号：Q959.837[生物学—动物学] Q78