| dc.description.abstract | Actinidin is an intracellular cysteine protease found in the fruit of Chinese
gooseberry (Actinidia chinensis) or Kiwi fruit. A variant of actinidin-encoding DNA
sequence has been constructed by amplifying a specific region of the full-length
actinidin cDNA using PCR method. Amplification was targeted to construct an
actinidin-encoding DNA sequence consisting of mature actinidin-encoding DNA plus
part of the N-terminal extension (carrying the QRTN motif) and part of the C-terminal
extension without QR sequence motif. The amplified fragment was then cloned in an
expression-secretion vector (pYSV9) and expressed in Saccharomyces cerevisiae.
The results of this study demonstrated that the cloned fragment had a smaller
size as compared to the PCR amplification product, suggesting that a rearrangement
event occured during the cloning process. Yeast culture supernatant was analysed for
expression of actinidin by using anti-actinidin polyclonal antibody raised in chicken.
The expression of the cloned fragment in S. cerevisiae, however, did not result in any
detectable expression product on the immunoblot. | en_US |
