Comparative analysis of genomic sequence and expression of a lipid transfer protein gene family in winter barley

Three genomic clones (gb/*4.2, gb/f4.6, gb/t4.9) have been isolatedfrom a barley [Hordeum vulgare L.) cv. Igri genomic libraryusing a cDNA clone of the low temperature responsive gene b/r4.1as the probe. The genomic clones are closely related and fromsequence homology are believed to be three further members ofa barley gene family encoding lipid transfer proteins. One clone,gb/t4.9, corresponds to a second cDNA clone (Jb/f4.9). Northernblot analysis revealed that the three genes reported here areinduced by low temperature and drought; however, the relativelevel of response to these stimuli was shown to differ betweenthe genes. Further, the developmental response of gb/?4.2 isdifferent from that found for other members of the gene family.The degree of low temperature induction of the blt4 gene familyappears to be cultivar-dependent. Comparative analysis of the putative promoter regions of thegenomic clones indicates three regions that show over 80 sequenceconservation between gb/f4.9 and gb/f4.6. Also identified areputative cis acting elements previously identified in ABA responsivegenes and low temperature inducible genes. The possible functionalsignificance of the conserved regions and elements is discussedin relation to the expression of the blt4 gene family. Key words: Barley, gene expression, genomic sequence, lipid transfer protein, low temperature     

Molecular evolution and structure--function relationships of the superoxide dismutase gene families in angiosperms and their relationship to other eukaryotic and prokaryotic superoxide dismutases

This study assesses whether the phylogenetic relationships between SODs from different organisms could assist in elucidating the functional relationships among these enzymes from evolutionarily distinct species. Phylogenetic trees and intron positions were compared to determine the relationships among these enzymes. Alignment of Cu/ZnSOD amino acid sequences indicates high homology among plant sequences, with some features that distinguish chloroplastic from cytosolic Cu/ZnSODs. Among eukaryotes, the plant SODs group together. Alignment of the Mn and FeSOD amino acid sequences indicates a higher degree of homology within the group of MnSODs (>70%) than within FeSODs (approximately 60%). Tree topologies are similar and reflect the taxonomic classification of the corresponding species. Intron number and position in the Cu/Zn Sod genes are highly conserved in plants. Genes encoding cytosolic SODs have seven introns and genes encoding chloroplastic SODs have eight introns, except the chloroplastic maize Sod1, which has seven. In Mn Sod genes the number and position of introns are highly conserved among plant species, but not among nonplant species. The link between the phylogenetic relationships and SOD functions remains unclear. Our findings suggest that the 5' region of these genes played a pivotal role in the evolution of function of these enzymes. Nevertheless, the system of SODs is highly structured and it is critical to understand the physiological differences between the SODs in response to different stresses in order to compare their functions and evolutionary history.     

Murine ferritin heavy chain: isolation and characterization of a functional gene

A mouse liver genomic library screened with a full-length cDNA encoding murine ferritin heavy chain (mFHC) [Torti et al., J. Biol. Chem. 263 (1988) 12638-12644] yielded a functional genomic clone mFHC. The genomic clone isolated included a region of approximately 3 kb containing four exons and three introns. Sequence comparisons of the mouse genomic clone with other genomic clones from rat, human and chicken showed a high degree of similarity among species in the coding regions. Introns and flanking sequences were less conserved. However, comparison of mFHC promoter elements with FHC genes from other species revealed common elements. Analysis of the genomic structure of FHC suggested the presence of pseudogenes. S1 nuclease analysis, however, confirmed that this mouse clone, when transfected into human MRC-5 fibroblasts, was transcribed, indicating that this clone contains an FHC functional gene.     

Expression of an Arabidopsis potassium channel gene in guard cells.

The Arabidopsis thaliana KAT1 cDNA encodes a voltage-gated inward-rectifying K+ channel. A KAT1 genomic DNA clone was isolated and sequenced, and a 5' promoter and coding sequences containing eight introns were identified. Reporter gene analysis of transgenic plants containing the KAT1 promoter fused to bacterial beta-glucuronidase showed robust beta-glucuronidase activity primarily in guard cells.