Reference "Molecular characterization of a gene for alanine aminotransferase from rice (Oryza sativa)"

Reference ID

2041

Title

Molecular characterization of a gene for alanine aminotransferase from rice (Oryza sativa)

Source

Plant molecular biology, 1999, vol. 39, pp. 149-159

Authors (5)

Kikuchi-H	Hirose-S
Toki-S	Akama-K
Takaiwa-F

Abstract

A cDNA clone encoding alanine aminotransferase (AlaAT) has isolated from
randomly sequenced clones derived from a cDNA library of maturing rice seeds by
comparison to previously identified genes. The deduced amino acid sequence was
88% and 91% homologous to those of the enzymes from barley and broomcorn millet
(Panicum miliaceum), respectively. Using this cDNA as a probe, we isolated and
sequenced the corresponding genomic clone. Comparison of the sequences of the
cDNA and the genomic gene revealed that the coding region of the gene was
interrupted by 14 introns 66 to 1547 bp long. Northern and western blotting
analyses showed that the gene was expressed at high levels in developing seeds.
When the 5'-flanking region between -930 and +85 from the site of initiation of
transcription was fused to a reporter gene for beta- glucuronidase (GUS) and
then introduced into the rice genome, histochemical staining revealed strong GUS
activity in the inner endosperm tissue of developing seeds and weak activity in
root tips. Similar tissue-specific expression was also detected by in situ
hybridization. These results suggest that AlaAT is involved in nitrogen
metabolism during the maturation of rice seed.