Nucleotide sequences of cDNA clones for C-hordein polypeptides

Abstract

Two partial cDNA clones, pc horl-3 and pc-919, encoding the carboxyterminal portion of two different C-hordein polypeptides have been sequenced, pc horl-3 can be translated into a frame of 74 amino acids, part of which is identical to the amino acid sequence of a C-hordein chymotryptic peptide. The 3' non-coding region is 174 nucleotides in length and extends to the polyadenylation site. The region has two putative polyadenylation signals, AATAAA, located 27 and 80 nucleotides upstream from the poly(A)-tail. Three RNA species, 1800, 2100 and 4300 bases in length, can be detected in the endosperm using pc hor 1-3 as probe. These could code for C-hordein polypeptides of 48, 67 and 144 kD, respectively. Hybridization of pc hor1-3 to restriction endonuclease fragments of nuclear barley DNA suggests that 6-8 genes code for the C-hordein polypeptides.The nucleotide sequence of the second cDNA clone, pc-919, can be translated into 64 amino acids, of which the 22 carboxyterminal amino acid residues are identical to the carboxyterminal reading frame encoded by pc horl-3. The adjacent 42 amino acid long reading frame shows homology to that deduced from pc hor 1-3, but the nucleotide sequence is different and highly homologous to the 5' coding sequence found in a B-hordein genomic clone. Hybridization using this part of pc-919 as a probe detects RNAs of 1200 and 1400 bases only. Thus pc-919 represents a gene containing sequences related to both Hor 1 and the Hor2 locus, and is the first nucleotide sequence data giving evidence that the two loci are related.

The prolamin storage proteins of cereal seeds: structure and evolution

“…4a, part a) have major components with Mr values of about 52000-54000 (as determined by sedimentation equilibrium ultracentrifugation [60] and intrinsic viscosity measurements in 5.9 M-guanidinium chloride [61]). Partial amino acid sequences determined directly [62][63][64] and deduced from cloned cDNAs [65,66] show unique N-terminal and C-terminal domains of 12 andK 6 residues respectively, flanking a repetitive domain calculated (from the Mr values) as consisting of over 400 residues (Fig. 4b).…”

Phytochrome Regulation of Greening in Barley

“…Subsequent isolation of RNA from the transcription mixture, preparation of plasmid DNA, dot blot hybridization, and quantitation of the results are also as described by Mosinger et al (18). In addition to probes for LHCP and Pchl-reductase mRNA species, a probe for cytoplasmic rRNA (11), the barley storage protein hordein (21), and the vector itself, pBR322, were also included as controls in all hybridizations of transcription products. Since in all cases equal counts were used for hybridization of the various samples, the changes in specific transcripts observed represent relative changes in the abundance of the various RNAs with respect to the total, rather than any absolute increases or decreases.…”

Mapping of theHor2 locus in barley by pulsed field gel electrophoresis

“…The 720 bp insert from pchor2-4, a B-hordein eDNA clone (36), cross-hybridizes to all Bhordein genes under the conditions described in materials and methods, and was therefore selected as a probe to identify the Hor2 locus. In order to identify individual fragments of the Hor2 .…”

“…The Hor2 locus has been estimated to contain at least 13 copies of B-hordein genes (25), accounting for the polymorphism in the polypeptides. Horl is less complex than Hor2 (35), while Hot3 only contains 1-2 genes. The genes encoding the ?-hordein polypeptides, a minor storage protein group (39), constitute another small multigenic family in barley (7).…”