Apr 8, 2020

Chlamydomonas reinhardtii Tubulin-Gene Disruptants for Efficient Isolation of Strains Bearing Novel Tubulin Mutations

BioRxiv : the Preprint Server for Biology
Takako Kato-MinouraR. Kamiya

Abstract

The single-cell green alga Chlamydomonas reinhardtii possesses two -tubulin genes (tua1 and tua2) and two {beta}-tubulin genes (tub1 and tub2), with the two genes in each pair encoding identical amino acid sequences. Here, we used an aphVIII gene cassette insertional library to establish eight disruptants with defective tua2, tub1, or tub2 expression. None of the disruptants exhibited apparent defects in cell growth, flagellar length, or flagellar regeneration after amputation. Because few tubulin mutants of C. reinhardtii have been reported to date, we then used our disruptants, together with a tua1 disruptant obtained from the Chlamydomonas Library Project (CLiP), to isolate novel tubulin-mutants resistant to the anti-tubulin agents propyzamide and oryzalin. As a result of several trials, we obtained 8 strains bearing 7 different a-tubulin mutations and 24 strains bearing 12 different b-tubulin mutations. Some of these mutations are known to confer drug resistance in human cancer cells. Thus, single-tubulin-gene disruptants are an efficient means of isolating novel C. reinhardtii tubulin mutants.

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Mentioned in this Paper

Biochemical Pathway
T-Lymphocyte
Virus
Genes
Virus Replication
Cell Culture Techniques
HIV Infections
IFNA2 gene
Obstruction
IFNA1 gene

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