Reference "Analysis of barley nitrate reductase cDNA and genomic clones"

Reference ID

8700

Title

Analysis of barley nitrate reductase cDNA and genomic clones

Source

Molecular & General Genetics, 1991, vol. 227, pp. 411-416

Authors (5)

Schnorr-K-M	Juricek-M
Huang-C-X	Culley-D
Kleinhofs-A

Abstract

Barley nitrate reductase cDNA and genomic clones were isolated by homology with
the barley nitrate reductase cDNA clone bNRp10 and sequenced. This is the first
reported analysis of a full-length nitrate reductase gene and its corresponding
cDNA in the same species. The longest cDNA clone extends to within 9 bp of the
ATG start codon and the sequence is similar to that reported for the higher
plant NR sequences. As expected, the amino acid sequence of barley nitrate
reductase is more related closely to the rice (84% homology) than to the
Arabidopsis (62%) sequence. Four different polyA addition sites were identified
from sequence analysis of nine barley NR cDNA clones. A 7.3 kb region of a
genomic recombinant lambda clone was subcloned as two contiguous BamHI fragments
into p Bluescript, designated pMJ7 and pMJ8, and sequenced. These clones include
the entire nitrate reductase coding region, one large intron, 2.7 kb of
untranslated sequence 5' to the translation start codon and 0.25 kb 3' to the
translation termination codon. The mRNA cap site was identified as a cytosine,
111 bases upstream of the ATG translation start codon. The putative CAAT and
TATA boxes were identified at -115 and -33 bp, respectively, with the mRNA cap
site designated as +1. The barley nitrate reductase gene coding region strongly
favors G or C in the third codon position.