摘要
采用小片段克隆法构建了方正银鲫[(Carassius auratus gibelio(Block)]部分基因组文库,并采用非影印的方法将转化子转移到硝酸纤维素滤膜上。用人工合成、并经放射性同位素[γ-32P]ATP标记的(CA)15探针对构建的基因组文库进行筛选,获得137个阳性克隆,经测序得到微卫星序列130个,其中完美型、非完美型、混合型分别占64.3%、16.9%和18.8%。应用引物设计软件Primer Primer5.0进行引物设计,得到引物103对。合成30对引物,随机选取其中5对进行PCR扩增。结果表明,这5对引物均可以稳定扩增。
The partial genome library of the silver crucian carp was constructed by method of small fragments DNA cloning and transferred to filter membrane by nonphotocopy method. The probe （CA）15 was synthesized and labeled with [γ-^32p] ATP,and was used to screen the library. One hundzed and thirty-seven positive clones were obtained. After sequencing, there are 130 microsatellite sequences and perfect, imperfect and compound ones take up 64.3 %, 16.9 % and 18.8 %, respectively. One hundred and three pairs of primer were designed with Primer Premier5.0. software and 30 pairs of them were synthesized. Five primers were selected randomly and amplified. The results show they can give good bands and the observed heterozygosity is high.
出处
《中国水产科学》
CAS
CSCD
北大核心
2006年第6期990-994,共5页
Journal of Fishery Sciences of China
基金
国家自然科学基金(30271010)
关键词
雌核发育
银鲫
微卫星DNA
gynogenetic
Carassius auratus gibelio
microsatellite DNA
