期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：1975
卷号：72
期号：3
页码：1072-1076
DOI：10.1073/pnas.72.3.1072
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：An in vitro system for the production of integrative recombinant DNA of bacteriophage lambda is described. The in vitro recombination mimics the in vivo integration of viral DNA into host DNA in its requirement for int gene product, for the presence of a thermolabile component, and for the limitation of the recombination to a pair of specialized sites (attachment sites) on the DNA. The enzymes are extracted from Escherichia coli containing phage lambda gene products. The substrate is the DNA from lambda-attB-attP, a phage variant that contains two attachment sites on the same chromosome. The product is a recombinant phage chromosome that has lost the DNA between the attachment sites. The parental and recombinant DNA are distinguished following transfection to mature phage in spheroplasts. The reaction requires ATP, Mg++, spermidine, and a monovalent cation. Recombination occurs preferentially between attachment sites on the same molecule. The enzymatic activity is completely inhibited by extracts containing xis gene product.
