S. Mano et al., STROMAL AND THYLAKOID-BOUND ASCORBATE PEROXIDASES ARE PRODUCED BY ALTERNATIVE SPLICING IN PUMPKIN, FEBS letters, 413(1), 1997, pp. 21-26
A cDNA encoding stromal ascorbate peroxidase (sAPX) was isolated using
poly(A)(+) RNA of pumpkin cotyledons by RT-PCR. The cDNA encodes a po
lypeptide with 372 amino acids and shares complete sequence identity w
ith pumpkin thylakoid-bound ascorbate peroxidase (tAPX), except for th
e deletion of a putative membrane spanning region located in the carbo
xy domain of tAPX. Southern blot hybridization and analysis of intron
structure indicated that mRNAs for sAPX and tAPX, whose suborganellar
localizations in chloroplasts are different, are produced by alternati
ve splicing, Immunoblot analysis showed that the accumulation of sAPX
and tAPX was differently regulated during germination and subsequent g
reening of pumpkin cotyledons. (C) 1997 Federation of European Biochem
ical Societies.