ArticlePloS one2013
SOMA: a single oligonucleotide mutagenesis and cloning approach.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
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- Cilia-localized GID/CTLH ubiquitin ligase complex regulates protein homeostasis of sonic hedgehog signaling components.Journal of cell science · 2022Article
- Post-transcriptional regulation of MRTF-A by miRNAs during myogenic differentiation of myoblasts.Nucleic acids research · 2020Article
- The GID ubiquitin ligase complex is a regulator of AMPK activity and organismal lifespan.Autophagy · 2020Article
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- Targeting of the AXL receptor tyrosine kinase by small molecule inhibitor leads to AXL cell surface accumulation by impairing the ubiquitin-dependent receptor degradation.Cell communication and signaling : CCS · 2019Article
- Regulation of MRTF-A by JMY via a nucleation-independent mechanism.Cell communication and signaling : CCS · 2018Article
- Ssy5 is a signaling serine protease that exhibits atypical biogenesis and marked S1 specificity.The Journal of biological chemistry · 2018Article
- Myocardin-Related Transcription Factor A Activation by Competition with WH2 Domain Proteins for Actin Binding.Molecular and cellular biology · 2016Article
- RMND5 from Xenopus laevis is an E3 ubiquitin-ligase and functions in early embryonic forebrain development.PloS one · 2015Article
- The conserved AAA-ATPase Msp1 confers organelle specificity to tail-anchored proteins.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
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5 authors.
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Abstract
Modern biology research requires simple techniques for efficient and restriction site-independent modification of genetic material. Classical cloning and mutagenesis strategies are limited by their dependency on restriction sites and the use of complementary primer pairs. Here, we describe the Single Oligonucleotide Mutagenesis and Cloning Approach (SOMA) that is independent of restriction sites and only requires a single mutagenic oligonucleotide to modify a plasmid. We demonstrate the broad application spectrum of SOMA with three examples. First, we present a novel plasmid that in a standardized and rapid fashion can be used as a template for SOMA to generate GFP-reporters. We successfully use such a reporter to assess the in vivo knock-down quality of morpholinos in Xenopus laevis embryos. In a second example, we show how to use a SOMA-based protocol for restriction-site independent cloning to generate chimeric proteins by domain swapping between the two human hRMD5a and hRMD5b isoforms. Last, we show that SOMA simplifies the generation of randomized single-site mutagenized gene libraries. As an example we random-mutagenize a single codon affecting the catalytic activity of the yeast Ssy5 endoprotease and identify a spectrum of tolerated and non-tolerated substitutions. Thus, SOMA represents a highly efficient alternative to classical cloning and mutagenesis strategies.
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