&CHAPTER 7

Recombinant DNA, Vector Design,

and Construction

MARK D. CURTIS

Institute of Plant Biology, University of Zurich, Switzerland

7.0. CHAPTER SUMMARY AND OBJECTIVES

7.0.1 Summary

Genomics, biotechnology, and biology in general have been enabled by methodologies to

manipulate DNA in a test tube (a very tiny test tube). Restriction enzymes are used as mol-

ecular scissors, and ligases are used as molecular “glue.” The polymerase chain reaction has

become invaluable in amplifying and cloning DNA. In addition, recombination systems

have been developed as alternatives to restriction enzymes as cloning tools. All these

methods are useful in creating plasmids containing chimeric DNA constructs that will be

transformed into plants.

7.0.2 Discussion Questions

1. What basic elements should be included in the design and construction of an efficient
   ubiquitous and constitutive plant gene expression vector?


2. Discuss the advantages and disadvantages of recombination cloning technologies
   versus traditional restriction digestion and ligation technology.


3. Describe a novel strategy to generate a T-DNA vector that allows the expression of
   several genes from a single position in the genome.


4. Discuss the advantages and disadvantages of using plastid vectors for plant trans-
   formation and gene expression.


5. Describe ways in which transgene technology could be made more acceptable to
   the public.

Plant Biotechnology and Genetics: Principles, Techniques, and Applications, Edited by C. Neal Stewart, Jr.

Copyright#2008 John Wiley & Sons, Inc.

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