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建立哺乳动物的基因文库是分离基因,研究基因结构和功能的重要条件之一。为了分离人食管癌的转化基因,我们首先建成了正常人食管粘膜组织的基因文库。 制备基因文库所用的载体是人噬菌体EMBL_3。这是一种在人1059载体的基础上改建而成的新载体,具有多种优点:1.去掉了人1059中所包含的Co1E1顺序,因而可用克隆到pBR322和其它Co1E1相关质粒中的DNA顺序作为探针,进行文库的筛选。2.在载体的两臂与中间片段之间,插入一段对称的多内切酶切点顺序。用双酶解的方法,可使载体中间片
The establishment of mammalian gene library is one of the important conditions for gene isolation and gene structure and function research. In order to isolate human esophageal cancer transformation genes, we first built a normal human esophageal mucosa gene library. The vector used to prepare the gene library was human bacteriophage EMBL_3. This is a new vector based on the human 1059 vector with a number of advantages: 1. The sequence of Co1E1 contained in human 1059 has been removed so that the DNA sequence cloned into pBR322 and other Co1E1-associated plasmids can be used As a probe, library screening was performed. 2. Insert a symmetrical sequence of multiple restriction endonucleases between the two arms of the vector and the middle segment. With double enzymatic method, the intermediate carrier can be made