Maniatis T, Hardison R C, Lacy E, Lauer J, O'Connell C, Quon D, Sim G K, Efstratiadis A
Cell. 1978 Oct;15(2):687-701. doi: 10.1016/0092-8674(78)90036-3.
We present a procedure for eucaryotic structural gene isolation which involves the construction and screening of cloned libraries of genomic DNA. Large random DNA fragments are joined to phage lambda vectors by using synthetic DNA linkers. The recombinant molecules are packaged into viable phage particles in vitro and amplified to establish a permanent library. We isolated structural genes together with their associated sequences from three libraries constructed from Drosophila, silkmoth and rabbit genomic DNA. In particular, we obtained a large number of phage recombinants bearing the chorion gene sequence from the silkmoth library and several independent clones of beta-globin genes from the rabbit library. Restriction mapping and hybridization studies reveal the presence of closely linked beta-globin genes.
我们介绍了一种真核生物结构基因分离方法，该方法涉及基因组DNA克隆文库的构建与筛选。通过使用合成DNA接头，将大的随机DNA片段连接到λ噬菌体载体上。重组分子在体外被包装成有活力的噬菌体颗粒并进行扩增，以建立永久性文库。我们从果蝇、家蚕和兔基因组DNA构建的三个文库中分离出了结构基因及其相关序列。特别地，我们从家蚕文库中获得了大量携带绒毛膜基因序列的噬菌体重组体，从兔文库中获得了几个β-珠蛋白基因的独立克隆。限制性图谱分析和杂交研究揭示了紧密连锁的β-珠蛋白基因的存在。
