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Transformation with AGL-1/pABr1 (Figure 1a) produced stable transformants with a strong and long-lasting SGFP expression (Figure 1b).
A maximum number of transformants with a minimal amount of DNA requires a high transformation efficiency.
When six compound leaves were grown, the plants were sprayed with 100 mL of spore suspension of the transformants with a concentration of 5 × 106 spores mL−1.
As shown in Figure 8A, seed viability and the time course of germination were similar for untransformed control and two transformants with a sharply different rhGCase content (63.5 and 215 mg per kg of flour).
Transformants with a targeted integration of the construct at the hacA locus were screened by Southern blot analysis.
We also isolated green fluorescent protein (GFP -expressing transformants with a nonhomoloGFP -expressingn of transformantsvector.
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Ethidium bromide-resistant transformants were selected in a single step, but at a transformation frequency of 10−6 to 10−7, 10- to 20-fold lower than that for transformants transformed with a spectinomycin resistance cassette under the same conditions.
As control, a sample of transformants with an altered morphology was purified through single conidial isolation.
Effective library sizes varied between 2.4 × 10 and 3.1 × 10 transformants with an average of 1.1 × 10 transformants.
However, the simultaneous insertion of extra copies of the crtYB and crtS (which encodes the astaxanthin synthase) genes in an astaxanthin-overproducing strain obtained by random mutagenesis resulted in transformants with an even higher astaxanthin content [ 20].
To achieve a steady photobiological hydrogen production, a screening protocol was designed to identify C. reinhardtii DNA insertional mutagenesis transformants with an attenuated photosynthesis to respiration capacity ratio (P/R ratio).
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