Subject culture-based methods

New Test Methods - A procedure has been established for treat-ing cutaneous Candida albicans infections of the rabbit triamcinolone acetonide (Kenalog Parenteral ) was used to depress host responses before and after localized infection of the ear lobe.15 Therapeutic efficacy of topically applied agents was based on a subjective evaluation of the lesions and culture scores. 

The berberine bridge enzyme utilized in tet-rahydroisoquinoline alkaloid pathways was cloned by this method [19]. This enzyme was purified from elicited Eschscholzia califomica cell suspension cultures in six to seven steps, where approximately 10 pg of pure protein was obtained from 700 mg of crude protein. The purified protein was digested with trypsin and the resulting peptides were subjected to N-terminal sequencing. An oligonucleotide primer based on one of these amino acid sequences was shown to hybridize to a clone Irom an E. califomica cDNA library. This clone was sequenced and shown to encode the amino acid sequences observed by N-terminal sequencing of the purified protein. The clone was then het-erologously expressed in S. cerevisea and was shown to catalyze the expected biochemical activity - formation of (5)-reticuline to (S)-scoulerine - as evidenced by a number of spectroscopic characterizations. 

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