Exploring gene function: use of yeast artificial chromosome transgenesis.
Transgenesis is a very powerful tool in functional analysis of proteins and control of gene expression. One of the main drawbacks has been the low levels of expression, lack of tissue specificity, and inappropriate expression frequently observed for transgenes made with small plasmid-based constructs. The use of much larger DNA fragments cloned in yeast artificial clones (YACs), bacterial artificial clones, or P1-based artificial clones has been found to give much better levels of expression, generally very close to that of an endogenous gene, and tissue-specific expression matching that of the endogenous gene. In addition, the large DNA can easily be subtly modified by homologous recombination. This article describes the background and methods of YAC transgenesis.
Huxley
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Department of Biochemistry and Molecular Genetics, Imperial College School of Medicine at St. Mary's, London, United Kingdom.
D000818:Animals; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D003470:Culture Media / Q000737:chemistry; D016521:Electrophoresis, Gel, Pulsed-Field; D015870:Gene Expression / Q000235:genetics; D018014:Gene Transfer Techniques; D005819:Genetic Markers / Q000235:genetics; D008561:Membrane Fusion / Q000502:physiology; D051379:Mice; D008822:Mice, Transgenic; D008845:Microinjections / Q000379:methods; D010714:Phosphatidylethanolamines / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic / Q000235:genetics; D015183:Restriction Mapping; D013234:Stem Cells / Q000378:metabolism
D003470:Culture Media; D005819:Genetic Markers; D010714:Phosphatidylethanolamines; C020888:1,2-dielaidoylphosphatidylethanolamine