Molecular cloning and in situ hybridization of alpha-l-arabinofuranosidase from carrot cells.
The cDNA of extracellular alpha-l-arabinofuranosidase (alpha-l-AFase, EC 3.2.1.55) secreted from suspension-cultured carrot cells (Daucus carota L. cv. Kintoki) was isolated and characterized. The nucleotide sequence of the cDNA (2.4 kb) revealed an open reading frame consisting of 655 amino acid residues. Sequence homology research showed 28.4% identity to the alpha-l-AFase A protein of Aspergillus niger. The genomic DNA was cloned by PCR, and the nucleotide ligature sequence showed 18 exons and 17 introns. The first intron was upstream of the initiation codon. In situ hybridization revealed that the alpha-l-AFase gene is expressed in the root meristem, elongation zone and the root hair of carrot seedlings, indicating that this enzyme may participate in cell proliferation and development of carrot root cells.
Tanaka
Reiji
R
Shionogi Aburahi Laboratories, 1405 Gotanda, Koka, Shiga, 520-3423, Japan Research Institute for Bioresources, Okayama University, Kurashiki, Okayama, 710-0046, Japan.
Ikeda
Minoru
M
Funatsuki
Kenzo
K
Yukioka
Hideo
H
Katoh
Kenji
K
Konno
Haruyoshi
H