Development of gene function analysis technology in the genus Neopyropia (transient expression)

It was found that for stable expression of the transgene in the cells of the rust fungus, it is important to use a transcription control region called a promoter derived from Neopyropia yezoensis (to promote transcription) and to optimize the DNA sequence of the transgene to match the codon of Neopyropia yezoensis gene (to promote translation).

Four gene constructs introduced into the cells of Neopyropia yezoensis

Description of the introduced gene constructs

35S:
A Promoter of CaMV35S virus, which infects land plants

GUS: β-glucuronidase
Reporter gene from E. coli. When expressed, it turns cells blue.

GAPDH:
Promoter (GAPDH gene) derived from Neopyropia yezoensis.

PyGUS:
Modified GUS (β-glucuronidase) whose DNA sequence is adapted to the Neopyropia yezoensis codon without changing the amino acid sequence.

By examining the number of GUS-expressing cells in the four transgenic constructs, we found that the 35S promoter, a promoter commonly used in land plants, is rarely functional and the E. coli gene is rarely translated in the Neopyropia yezoensis cells.