摘要
为筛选、克隆泥鳅性别特异基因奠定基础,以黄河中下游雌性和雄性成体泥鳅为材料,采用抑制性消减杂交技术(SSH)构建基因组DNA文库,并结合蓝白斑筛选和PCR方法鉴定文库质量。结果表明:雌雄泥鳅基因组DNA的正反向抑制消减文库构建成功,其中,正向文库包含240个克隆,反向文库包含216个克隆,阳性克隆率为95.61%,插入片段范围为0.2～1.0kb。
In order to provide a basis for screening and cloning the gender-specific genes,the genomic DNA library was constructed by suppression subtractive hybridization（SSH） technique taking M.anguillicaudatus as experimental materials.And blue-white blot and PCR were used to detect the subtracted DNA library quality.The results showed that the forward and reverse suppression subtracted genomic DNA library was successfully constructed,the forward library contained 240 clones and 216 clones in the reverse library,the positive ratio of constructed genomic DNA libraries was 95.61% and the length of DNA fragments ranged from 0.2 to 1.0 kp.
出处
《贵州农业科学》
CAS
北大核心
2012年第10期141-144,共4页
Guizhou Agricultural Sciences
基金
河南师范大学引进人才科研项目"DMRT1基因不同剪接体的克隆和时空表达模式分析"(01046500109)
关键词
泥鳅
抑制性消减杂交(SSH)
基因组DNA文库
性别特异基因
Misgurnus anguillicaudatus
suppression subtractive hybridization（SSH）
genomic DNA library
gender-specific genes
