ArticlePlant methods2017
A robust protocol for efficient generation, and genomic characterization of insertional mutants of
Article in Plant methods, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 21 citations in OpenAlex.
- Increasing lipid production inFrontiers in bioengineering and biotechnology · 2024Article
- How abiotic stress-induced socialization leads to the formation of massive aggregates in Chlamydomonas.Plant physiology · 2022Article
- Discovery of photosynthesis genes through whole-genome sequencing of acetate-requiring mutants of Chlamydomonas reinhardtii.PLoS genetics · 2021Article
- Article
- Simple innovative adaptor to improve genome walking with convenient PCR.Journal, genetic engineering & biotechnology · 2020Article
- Metabolic engineering of ketocarotenoids biosynthetic pathway in Chlamydomonas reinhardtii strain CC-4102.Scientific reports · 2020Article
- TIM, a targeted insertional mutagenesis method utilizing CRISPR/Cas9 in Chlamydomonas reinhardtii.PloS one · 2020Article
- MAPINS, a Highly Efficient Detection Method That Identifies Insertional Mutations and Complex DNA Rearrangements.Plant physiology · 2018Article
- Identification and characterization of a solute carrier, CIA8, involved in inorganic carbon acclimation in Chlamydomonas reinhardtii.Journal of experimental botany · 2017Article
- Efficient and stable transformation ofBioImpacts : BI · 2017Article
Corrections and comments
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Authors and funding
7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundRandom insertional mutagenesis of
resultsInsertional mutants were generated using a DNA cassette that confers paromomycin resistance. This protocol identified the cassette insertion site for greater than 80% of the transformants. In the majority of cases the insertion event was found to be simple, without large deletions of flanking genomic DNA. Multiple insertions were observed in less than 10% of recovered transformants.
conclusionThe method is quick, relatively inexpensive and does not require any special equipment beyond an electroporator. The protocol was tailored to ensure that the sequence of the Chlamydomonas genomic DNA flanking the random insertion is consistently obtained in a high proportion of transformants. A detailed protocol is presented to aid in the experimental design and implementation of mutant screens in Chlamydomonas.
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Registered trials
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