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The transformants were streaked on the SD/-Trp or SD/-Ade/-His/-Leu/-Trp medium supplemented with different concentration of 3-AT (0, 5 and 15 mM) and incubated at 30°C for 4 d, followed by evaluation of growth status of the transformants.

A β-glucan plate assay was used for preliminary evaluation of enzyme production by the transformants and for selection of representative clones from the transformation for further analysis.

This was confirmed through functional evaluation of their silenced transformants.

For easy evaluation of transactivation activity, transformants were dissolved in sterile distilled water and 10-fold serial dilutions prepared.

A majority of the transformants resistant to the antibiotic G418 were capable of synthesizing and secreting biologically active human interferon.

Multi-copy integration transformants had higher lipid contents than mono-copy integration transformants; the lipid content of the transformants was consistent with the copy number of acl gene integrated.

Next, we analyzed the growth of the transformants in liquid media using a Cell Growth Quantifier (Aquila Biolabs)39.

Southern blot analysis suggested that 87.5 % of transformants obtained with this protocol displayed single hybridization bands, indicating a single insert of T-DNA in each of the transformants.

Over 97% of the transformants retained the sulfonylurea resistance phenotype under non-selective conditions.

Although several of the vector-gpt DNAs can replicate in monkey and mouse cells, none of the transformants contain autonomously replicating vector-gpt DNA.

Intracellular growth of the transformants into free-living amoeba and murine and human macrophages decreased.