毛足棒角蝗基因组DNA文库的构建

Construction of a Genomic DNA Library of the Locust,Dasyhippus barbipes

摘要: 纯化毛足棒角蝗（Dasyhi ppus barbipes）虫体组织的高分子量基因组DNA，经限制性内切酶Sau3AI部分消化后，10%-40%蔗糖密度梯度离心分离10-20 kb的DNA片段。以λEMBL3为克隆载体，与10-20 kb的DNA片段进行连接和包装，构建了毛足棒角蝗的基因组DNA文库。经测定该文库的滴度是2×105 pfu/mL，根据计算，99%的毛足棒角蝗的基因包含在该基因组文库中。

Abstract: High molecular weight genomic DNA from the grasshopper,Dasyhippus barbipes was extracted and partially digested with Sau3AI,10-20 kb DNA fragments were collected by 10%-40% sucrose density gradient centrifugation,and ligated into lambda EMBL3 vector. Recombinant DNA was packaged in vitro,then construction of the genomic DNA library in Dasyhippus barbipes was completed.The titer of the library is 2× 105 pfu/mL,and the genomic DNA library consists of 99% genes of Dasyhippus barbipes.