貉微卫星DNA富集文库的构建与鉴定  

Construction and Identification of DNA Libraries Enriched for Microsatellite Repeat Sequences of Raccoon Dog

作　　者：闫守庆[1] 李冰[1] 祝万菊[2] 

机构地区：[1]吉林大学,长春130062 [2]吉林大学畜牧兽医学院

出　　处：《东北林业大学学报》2008年第6期48-50,共3页Journal of Northeast Forestry University

基　　金：吉林大学青年教师基金资助

摘　　要：貉基因组DNA经限制性内切酶MboI酶切,用2.0%琼脂糖凝胶电泳回收200～800bp的DNA片段,与MboI连接头连接,连接产物与生素物标记的(AC)n微卫星探针变性及退火,再通过链亲和素偶联磁珠亲和捕捉,经吸附、洗涤及洗脱,然后以洗脱产物为模板,通过PCR扩增,与pUCm-T载体连接,转化大肠杆菌DH5α,构建貉(AC)n微卫星DNA富集文库,经测序分析表明该文库含重组克隆约2800个,其中含有(AC)n微卫星DNA序列的阳性克隆占71.4%。Genomic DNAs from raccoon dog were cleaved with restriction enzymes MboI. The fragments in size from 200 bp to 800 bp were recovered by 2.0% agarose gel electrophoresis and ligated with adaptor, and then denatured and hybridized to a biotinylated （AC）n oligonucleotide. The biotinylated hybrids were retained on the magnetic particles according to the strong affinity between biotin and strept-avidin. The eluate was amplified by PCR and cloned into pUCm-T plasmid vector, and then transformed into Escherichia c.oli DH5α to construct DNA libraries enriched for microsatellite repeat sequences of raccoon dog. Sequence analysis shows that the library contains 2 800 clones and 71.4% of the clones contain （AC）n repeats.

关 键 词：貉 微卫星 遗传标记 

分 类 号：Q959.838[生物学—动物学]