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Fifteen and 23 transformants were obtained from transformation with the constructs.

More than 100 positive transformants were obtained after transforming the silencing construct into C. reinhardtii CC425.

Using this technique, up to 145 transformants were obtained per transformation plate, and a ku70 deletion strain was constructed in the ATCC 42464 background and characterized.

During development of the transformation protocol, transformation parameters were modified until paromomycin-resistant transformants were obtained (see below).

The six different modified 35S constructs were introduced through Agrobacterium-mediated transformation and 194 lines of independent transformants were obtained.

Even, no transformants were obtained when plasmid p602/22lux was transformed into B. subtilis BR151.

About 60 transformants were obtained for each of the transformations, ∼95% of which clearly showed the deletion phenotype (data not shown).

A total of ∼280 erythromycin (Erm -resistant transformants wErm -resistantn each of transformantsdent transformation experiments were strain 13.

The recombinant plasmids pPIC9K- BlAmy-opt and pPIC9K- BlAmy were linearized and transformed into P. pastoris GS115 and several thousands of transformants were obtained on MD plates.

Importantly, no transformants were obtained with unmethylated plasmid, validating the necessity to protect transforming DNA against the endogenous CpaAI restriction endonuclease.

Transformation of JWCH006 with methylated pJGW07 was successful, and uracil prototrophic transformants were obtained at a frequency of 22 ± 10 CFU per microgram of DNA (Table  2).