ABSTRACT
This chapter reviews the techniques for preparation of P1 DNA for transgenic expression studies, using the human apo-B gene and the human apoE/CI/CIV/CII gene cluster. For many years, investigators using transgenic animals have learned to live with the inherent limitations of these cloning vectors by constructing more compact “minigene” constructs from cDNA and genomic clones. However, minigene constructs invariably force the investigator to make assumptions about the regions of a gene. For studies of protein structure/function and for studies of gene regulation, it would be desirable to generate transgenic mice with genetically modified P1 clones. Sternberg reported methods for interrupting P1 clones with Tn10-based transposons and suggested that this technique would be useful for generating truncated proteins for structure/function studies. The chapter reviews the techniques for purifying the DNA from P1 clones and showed that fragments as large as 91 kb can be used to achieve high levels of expression in both transgenic mice and transgenic rabbits.
