第5章DNA导入活细胞.pptVIP

Chapter 5 Introduction of DNA into living cells contents 5.1 Transformation-the uptake of DNA by bacterial cells 5.1.1 Not all species of bacteria are equally efficient at DNA uptake 5.1.2 Preparation of competent E.coli cells 5.1.3 Selection for transformed cells 5.2 Identification of recombinants 5.2.1 Recombinant selection with pBR322-insertional inactivation of an antibiotic resistance gene 5.2.2 Insertional inactivation does not always involve antibiotic resistance contents 5.3 Introduction of phage DNA into bacterial cells 5.3.1 Transfection 5.3.2 In vitro packaging 5.3.3 Phage infection is visualized as plaques on an agar medium 5.4 Identification of recombinant phages 5.4.1 Insertional inactivation of a lacZ’ gene carried by the phage vector 5.4.2 Insertional inactivation of the ? cI gene 5.4.3 Selection using the Spi phenotype 5.4.4 Selection on the basis of ? genome size 5.5 Transformation of non-bacterial cells 5.5.1 Transformation of individual cells 5.5.2 Transformation of whole organisms Two purposes of cloning克隆的两个目的 To get a large number of recombinant DNA Purification Ligation production 连接产物 Unligated vector molecules Unligated DNA fragments Self-ligated vector Recombinant DNA carrying the wrong inserted DNA The desired recombinant DNA molecule Fig 5.2 Cloning is analogous to purification. From a mixture of different molecules, clones containing copies of just one molecule can be obtained. 5.1 Transformation-the uptake of DNA by bacterial cells转化-使细菌细胞获取DNA Transformation (转化): the introduction of any DNA molecule into any living cell. Competent cell(感受态细胞): A culture of bacteria that has been treated to enhance their ability to take up DNA molecules. Reception cell受体细胞 原核生物受体细胞——主要是大肠杆菌 另有：枯草杆菌