Site-Directed Mutagenesis by Fusion of Contiguous DNA Fragments

Carmona, Sergio; Passman, Marc; Kew, Michael C.; Arbuthnot, Patrick

doi:10.2144/99263bm02

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Site-directed Mutagenesis1

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2002

2003

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“…Altered sequences were created according to the method described by Carmona et al (24) with some modi®cations. Sequencing primers, M13 forward and backward, for the pGEM-3Z vector were used as the¯anking primers with the internal mutagenic (site-mutated) primers to introduce mutations into the sequences #1-3 and #1-6.…”

Section: Site-directed Mutagenesismentioning

confidence: 99%

Specific SR protein-dependent splicing substrates identified through genomic SELEX

Kim

Shi

Lee

et al. 2003

Nucleic Acids Research

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The Drosophila pre-mRNA splicing factor B52 (SRp55) is essential for fly development, but splicing of RNAs of specific genes tested previously is normal in B52-null animals, presumably due to partial functional redundancy with other SR proteins. To identify B52-dependent splicing substrates in vivo, we selected genomic sequence fragments whose transcripts bind B52. Almost all of the corresponding genes having a known function encode either transcription factors or components of signal transduction pathways, with the B52- binding fragments located to not only exonic but also intronic regions. Some pre-mRNAs from these genes showed splicing defects in the B52-null mutant. These results indicate that B52 has unique functions in the removal of some introns during development, and plays a critical role in cellular regulatory networks.

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Section: Site-directed Mutagenesismentioning

confidence: 99%