Chapter title |
From Selective Full-Length Genes Isolation by TAR Cloning in Yeast to Their Expression from HAC Vectors in Human Cells.
|
---|---|
Chapter number | 1 |
Book title |
Bacterial Artificial Chromosomes
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-1652-8_1 |
Pubmed ID | |
Book ISBNs |
978-1-4939-1651-1, 978-1-4939-1652-8
|
Authors |
Natalay Kouprina, Nicholas C O Lee, Artem V Kononenko, Alexander Samoshkin, Vladimir Larionov, Nicholas C. O. Lee, Artem V. Kononenko, Kouprina, Natalay, Lee, Nicholas C. O., Kononenko, Artem V., Samoshkin, Alexander, Larionov, Vladimir |
Abstract |
Transformation-associated recombination (TAR) cloning allows selective isolation of full-length genes and genomic loci as large circular Yeast Artificial Chromosomes (YACs) in yeast. The method has a broad application for structural and functional genomics, long-range haplotyping, characterization of chromosomal rearrangements, and evolutionary studies. In this paper, we describe a basic protocol for gene isolation by TAR as well as a method to convert TAR isolates into Bacterial Artificial Chromosomes (BACs) using a retrofitting vector. The retrofitting vector contains a 3' HPRT-loxP cassette to allow subsequent gene loading into a unique loxP site of the HAC-based (Human Artificial Chromosome) gene delivery vector. The benefit of combining the TAR gene cloning technology with the HAC gene delivery system for gene expression studies is discussed. |
Mendeley readers
Geographical breakdown
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Unknown | 16 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
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Researcher | 6 | 38% |
Student > Master | 4 | 25% |
Other | 1 | 6% |
Student > Bachelor | 1 | 6% |
Professor > Associate Professor | 1 | 6% |
Other | 1 | 6% |
Unknown | 2 | 13% |
Readers by discipline | Count | As % |
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Biochemistry, Genetics and Molecular Biology | 5 | 31% |
Chemical Engineering | 1 | 6% |
Immunology and Microbiology | 1 | 6% |
Unknown | 2 | 13% |