Building a multipurpose insertional mutant library for forward and reverse genetics in Chlamydomonas.

ABSTRACT: BACKGROUND:The unicellular green alga, Chlamydomonas reinhardtii, is a classic model for studying flagella and biofuel. However, precise gene editing, such as Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas9) system, is not widely used in this organism. Screening of random insertional mutant libraries by polymerase chain reaction provides an alternate strategy to obtain null mutants of individual gene. But building, screening, and maintaining such a library was time-consuming and expensive. RESULTS:By selecting a suitable parental strain, keeping individual mutants using the agar plate, and designing an insertion cassette-specific primer for library screening, we successfully generated and maintained ~150,000 insertional mutants of Chla

SUBMITTER: Cheng X 

PROVIDER: S-EPMC5430608 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature