Methods in Molecular Biology

Methods and Protocols

By: Andrew Ward (Editor)

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Hardcover | 10 October 2001

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390 Pages

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Genomic imprinting is the process by which gene activity is regulated according to parent of origin. Usually, this means that either the maternally inherited or the paternally inherited allele of a gene is expressed while the opposite allele is repressed. The phenomenon is largely restricted to mammals and flowering plants and was first recognized at the level of whole genomes. Nuclear transplantation experiments carried out in mice in the late 1970s established the non-equivalence of the maternal and paternal genomes in mammals, and a similar conclusion was drawn from studies of interploidy crosses of flowering plants that extend back to at least the 1930s. Further mouse genetic studies, involving animals carrying balanced translocations (reviewed in Chapter 3), indicated that imprinted genes were likely to be widely scattered and would form a minority within the mammalian genome. The first imprinted genes were identified in the early 1990s; over forty are now known in mammals and the list continues steadily to expand.

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"...a multi-authored volume in which the methods of gene imprinting are discussed in detail in 25 chapters. The protocols are very detailed with multiple schemes and figures." - Journal of Pediatric Endocrinology and Metabolism

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Non-Fiction Science Biology, Life Sciences Life Sciences in General Genetics excluding Medical Molecular Biology Science in General Medicine Pre-Clinical Medicine & Basic Sciences Medical Genetics

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Published: 10th November 2010

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Preface	p. v
Contributors	p. ix
Generation of Monoparental Embryos for Investigation into Genomic Imprinting	p. 1
Deriving and Propagating Mouse Embryonic Stem Cell Lines for Studying Genomic Imprinting	p. 21
Balanced Translocations for the Analysis of Imprinted Regions of the Mouse Genome	p. 41
Production of YAC Transgenic Mice by Pronuclear Injection	p. 55
A Transgenic Approach to Studying Imprinted Genes: Modified BACs and PACs	p. 67
Methylation-Sensitive Genome Scanning	p. 83
Subtraction-Hybridization Method for the Identification of Imprinted Genes	p. 101
Identification of Imprinted Loci by Methylation: Use of Methylation-Sensitive Representational Difference Analysis (Me-RDA)	p. 113
Ribonuclease Protection	p. 133
Quantitative RT-PCR-Based Analysis of Allele-Specific Gene Expression	p. 145
Allele-Specific In Situ Hybridization (ASISH)	p. 153
RNA-FISH to Analyze Allele-Specific Expression	p. 169
Flow Cytometry and FISH to Investigate Allele-Specific Replication Timing and Homologous Association of Imprinted Chromosomes	p. 181
Southern Analysis Using Methyl-Sensitive Restriction Enzymes	p. 193
A PCR-Based Method for Studying DNA Methylation	p. 205
Bisulfite-Based Methylation Analysis of Imprinted Genes	p. 217
Direct Analysis of Chromosome Methylation	p. 229
In Vitro Methylation of Predetermined Regions in Recombinant DNA Constructs	p. 243
In Vitro Methylation of Specific Regions in Recombinant DNA Constructs by Excision and Religation	p. 251
Detection of Methyl-Sensitive DNA-Binding Proteins with Possible Involvement in the Imprinting Phenomenon	p. 259
Probing Chromatin Structure with Nuclease Sensitivity Assays	p. 269
Examining Histone Acetylation at Specific Genomic Regions	p. 285
Purification of the McCP2/Histone Deacetylase Complex from Xenopus Iaevis	p. 297
Reconstitution of Chromatin In Vitro	p. 309
Genomic Imprinting in Plants	p. 327
Index	p. 371
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Methods in Molecular Biology

Methods and Protocols

At a Glance

Hardcover

Industry Reviews

Shipping

How to return your order

Defective items

You Can Find This Book In

Other Editions and Formats

Paperback

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