By Klemens J. Hertel
Providing a consultant to classical experimental methods to decipher splicing mechanisms and experimental options that depend on novel multi-disciplinary methods, Spliceosomal Pre-mRNA Splicing: equipment and Protocols describes the speculation of different pre-mRNA splicing in seven introductory chapters after which introduces protocols and their theoretical history correct for numerous experimental learn. those protocol chapters conceal easy easy methods to become aware of splicing occasions, analyses of different pre-mRNA splicing in vitro and in vivo manipulation of splicing occasions and high-throughput and bioinformatic analyses of other splicing. Written within the hugely winning Methods in Molecular Biology sequence structure, chapters contain introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, effectively reproducible protocols and tips about troubleshooting and fending off identified pitfalls.
Comprehensive and useful, Spliceosomal Pre-mRNA Splicing: tools and Protocols will relief newbies and professional molecular biologists in realizing the attention-grabbing global of other splicing with the last word target of paving the best way for plenty of new discoveries to come.
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Spliceosomal Pre-mRNA Splicing: Methods and Protocols
Offering a advisor to classical experimental methods to decipher splicing mechanisms and experimental recommendations that depend upon novel multi-disciplinary techniques, Spliceosomal Pre-mRNA Splicing: equipment and Protocols describes the speculation of different pre-mRNA splicing in seven introductory chapters after which introduces protocols and their theoretical historical past suitable for various experimental learn.
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Extra resources for Spliceosomal Pre-mRNA Splicing: Methods and Protocols
Example text
Both reason and evidence suggest that AS would be facilitated by a variety of features of animals’ intron–exon structures: (1) Large numbers of introns provide many opportunities for AS. (2) Heterogeneous intron boundaries, with associated differences in the strength of base pairing with the spliceosomal RNAs, allow for the possibility of regions for which recognition by the spliceosome might be “borderline”—leading to non-constitutive splicing of these regions. (3) Utilization of a variety of heterogeneous splicing signals—exonic and intronic splicing regulators, in addition to core splicing signals—allows for the possibility of regulating local splicing by regulation of the splicing factors that bind subsets of these signals.
In total, then, comparative studies of intronic and exonic sequences over long evolutionary distances within eukaryotes support a model in which ancestral eukaryotes had “animal-like” intron–exon structures, with frequent introns spliced by use of a combination of diffuse motifs including frequent ESEs and heterogeneous core splicing motifs. Over the course of evolution, many lineages have changed significantly, shedding the vast majority of their introns, evolving homogeneous core splicing motifs, and significantly decreasing dependence on auxiliary splicing motifs such as ESEs.
However, as discussed above, genomic-era studies have shown that the story is quite different from this: many of the features associated with AS in animals—frequent introns, heterogeneous splicing boundaries, introns with lengths exceeding “minimal” intron lengths, and utilization of auxiliary splicing signals—are not specific to animals, but are in fact quite common in modern eukaryotes as well as characteristic of eukaryotic ancestors [22]. Thus, the hypothesis that widespread productive AS in animals is “due” to these features, a hypothesis still commonly invoked in passing in publications, is strongly rejected, since these features are common in organisms with little or no productive AS.