Restriction enzymes construction

Liu, Q Li, M. Z., Leibham, D., Cortez, D., and Elledge, S. J. (1998). The univector plasmid-fusion system, a method for rapid construction of recombinant DNA without restriction enzymes. Current Biology 8, 1300-1309. 

Figure 9.8 The construction of restriction maps. The relative position of cleavage sites for two hypothetic restriction enzymes is illustrated for a 12 kb piece of DNA.

In your notebook, invent your own restriction enzyme. Show the DNA base sequence where it cleaves. Then construct a double stranded section of DNA with at least 25 base pairs and mark the points of cleavage by your enzyme. Finally, show all fragments formed. 

Workers in the early 1970s recognized that restriction enzymes provided tools not only for DNA mapping but also for construction of new DNA species not found in nature. A collection of recombinant DNA species consisting of many passenger sequences joined to identical vector molecules is called a library. Individual recombinant DNAs are isolated from single clones of the library for detailed analysis and manipulation. 

Electrophoretic analyses of restriction enzyme digests and construction of a restriction enzyme map (see Experiment 15)... 

Restriction endonucleases catalyze the hydrolysis of specific phosphodi-ester bonds in double-stranded DNA. These enzymes are often used to linearize a circular plasmid for hybrid DNA construction. They have also found use in the analysis of DNA and the construction of restriction maps. In this experiment, students will incubate various restriction enzymes with plasmid or viral DNA and analyze the product DNA fragments by agarose gel electrophoresis. 

Experiment 14 introduced students to some principles and techniques involved in recombinant DNA research. Specifically, the experiment outlined the replication, isolation, and analysis of bacterial plasmid vehicles for molecular cloning experiments. This experiment describes another tool that is essential in hybrid plasmid construction and analysis-restriction enzyme action. The procedures introduced here have also found widespread use in the analysis and characterization of all DNA molecules. 

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