乌金猪基因组文库的构建

Construction of Genomic Library of Wujin Pig

为探究与我国优良地方猪种乌金猪特性相关的基因机制,拟构建其基因组文库。取乌金猪肝脏组织,提取大小50 kb以上的基因组DNA,利用限制性内切酶Bcl I对基因组DNA进行随机酶切和低熔点琼脂糖电泳方法回收10～23 kb的DNA片段。以EMBL3作为载体,经BamH I酶切和去磷酸化处理后,与上述纯化的目的 DNA片段连接,在体外包装成重组噬菌体,重组噬菌体转染宿主菌KW251,构建成乌金猪基因组文库。文库的效价为2.4×109pfu/mL。乌金猪基因组文库的成功构建,为进行其相关功能基因和基因组区域的识别,基因组DNA和调控元件的克隆与功能分析等后续研究奠定了良好的基础。

To identify the genomic mechanisms for specific traits of Wujin pig, a domestic Chi- nese swine breed, we constructed its genomic library. The genomic DNA larger than 50 kb was extracted from Wujin pig liver tissue and was digested randomly by restriction enzyme Bcl I. DNA fragments ranging from 10 to 23 kb were recovered by agarose gel electrophoresis. EMBL3 vector was digested by BamH I, treated with calf intestine alkaline phosphatase, and then ligated with purified DNA fragments forementioned. The genomic library of Wujin pig was constructed by packing recombinant DNA in vitro with phage package protein and transecting KW251 host cells. The result showed that the titer of the library was 2.4 × 10^9 pfu/mL. The genomic li- brary of Wujin pig could be used for identification of genes and genomic regions of interest, and provide valuable data for further study such as cloning of genomic DNA and functional analysis of regulatory element.