植物科学

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PEG-mediated, Stable, Nuclear and Chloroplast Transformation of Cyanidioschizon merolae

PEG介导的稳定的Cyanidioschizon merolae细胞核和叶绿体转化

Maksymilian Zienkiewicz* TK Tomasz Krupnik AD Anna Drożak KK Kinga Kania

0 Q&A 3590 Views Sep 5, 2019

The ability to achieve nuclear or chloroplast transformation in plants has been a long standing goal, especially in microalgae research. Over past years there has been only little success, but transient and stable nuclear transformation has been achieved in multiple species. Our newly developed method allows for relatively simple transformation of Cyanidioschizon merolae in both nuclear and chloroplast genome by means of homologous recombination between the genome and a transformation vector. The use of chloramphenicol resistance gene as the selectable marker allows for plate-based efficient selection of mutant colonies. Overall, the method allows the generation of mutant strains within 6 months.

Preparation of Knockdown Transformants of Unicellular Charophycean Alga, Closterium peracerosum-strigosum-littorale Complex

轮藻单细胞，新月藻属peracerosum-strigosum-littorale复合体的基因沉默转化子的制备

JA Jun Abe NH Naoko Hirano AK Ayumi Komiya NK Naho Kanda AF Anri Fujiwara SH Sachie Hori YT Yuki Tsuchikane HS Hiroyuki Sekimoto*

0 Q&A 8968 Views May 20, 2016

To prepare the knockdown transformants of the Closterium peracerosum-strigosum-littorale (C. psl.) complex, particle bombardment was applied with a newly constructed vector (pSA0104) with an endogenous constitutive promoter fused to a DNA fragment corresponding to an antisense strand of a target gene. Using a hygromycin resistance gene (aph7”), hygromycin-resistant colonies were selected. After the second screening, integration of the vector into the genome was checked by PCR and the knockdown effect was evaluated by Western blotting using a specific antibody against the target protein.