Sequence analysis and regulation of expression of a gene coding for carbonic anhydrase in Chlamydomonas reinhardtii

1991 ◽  
Vol 69 (5) ◽  
pp. 1097-1102 ◽  
Author(s):  
J. R. Coleman ◽  
Irene Luinenburg ◽  
Nathalie Majeau ◽  
Nicholas Provart
Keyword(s):  
Sequence Analysis ◽  
Carbonic Anhydrase ◽  
Chlamydomonas Reinhardtii ◽  
Genomic Dna ◽  
Immunogold Labelling ◽  
Gene Organization ◽  
Amino Acid Sequences ◽  
Higher Plant ◽  
Regulation Of Expression ◽  
Dna Fragment

Using immunological and molecular biological techniques, we studied the localization, gene organization, and regulation of expression of the extracellular carbonic anhydrase in the eukaryotic green alga Chlamydomonas reinhardtii. Electron microscopy data using immunogold labelling suggest an association of the protein with the inner face of the cell wall. The same polyclonal antibody was previously used to select an immunoreactive 2.5-kb genomic DNA fragment coding for a portion of the carbonic anhydrase monomer. Using the known carbonic anhydrase cDNA sequence and sequence analysis of the genomic DNA fragment, we have deduced the exon–intron organization of the genomic clone. The similarities between Chlamydomonas and higher plant carbonic anhydrase amino acid sequences and the effect of photoheterotrophic growth on the expression of the algal carbonic anhydrase are also examined. Key words: carbonic anhydrase, Chlamydomonas, immunogold labelling, sequence, expression.

Cloning and sequence analysis of the genomic DNA fragment encoding oryzacystatin

Gene ◽  
1989 ◽  
Vol 81 (2) ◽  
pp. 259-265 ◽  
Author(s):  
Kendo Hiroto ◽  
Emori Yasufumi ◽  
Abe Keiko ◽  
Suzuki Koichi ◽  
Arai Soichi
Keyword(s):  
Sequence Analysis ◽  
Genomic Dna ◽  
Dna Fragment

Isolation and sequence analysis of the genomic DNA fragment encoding an aspartic proteinase inhibitor homologue from potato (Solanum tuberosum L.)

1992 ◽  
Vol 20 (2) ◽  
pp. 311-313 ◽  
Author(s):  
Darja Barlič Maganja ◽  
Borut Štrukelj ◽  
Jože Pungerčar ◽  
Franc Gubenšek ◽  
Vito Turk ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Sequence Analysis ◽  
Proteinase Inhibitor ◽  
Genomic Dna ◽  
Solanum Tuberosum L ◽  
Aspartic Proteinase ◽  
Dna Fragment

Molecular Markers for Rapidly Identifying Candidate Genes in Chlamydomonas reinhardtii: ERY1 and ERY2 Encode Chloroplast Ribosomal Proteins

Genetics ◽  
2003 ◽  
Vol 164 (4) ◽  
pp. 1345-1353
Author(s):  
Amber K Bowers ◽  
Jennifer A Keller ◽  
Susan K Dutcher
Keyword(s):  
Molecular Markers ◽  
Chlamydomonas Reinhardtii ◽  
Candidate Genes ◽  
Splice Site ◽  
Genomic Dna ◽  
Ribosomal Proteins ◽  
Genomic Sequence ◽  
Point Mutations ◽  
Acceptor Site ◽  
Wild Type

Abstract To take advantage of available expressed sequence tags and genomic sequence, we have developed 64 PCR-based molecular markers in Chlamydomonas reinhardtii that map to the 17 linkage groups. These markers will allow the rapid association of a candidate gene sequence with previously identified mutations. As proof of principle, we have identified the genes encoded by the ERY1 and ERY2 loci. Mendelian mutations that confer resistance to erythromycin define three unlinked nuclear loci in C. reinhardtii. Candidate genes ribosomal protein L4 (RPL4) and L22 (RPL22) are tightly linked to the ERY1 locus and ERY2 locus, respectively. Genomic DNA for RPL4 from wild type and five mutant ery1 alleles was amplified and sequenced and three different point mutations were found. Two different glycine residues (G102 and G112) are replaced by aspartic acid and both are in the unstructured region of RPL4 that lines the peptide exit tunnel of the chloroplast ribosome. The other two alleles change a splice site acceptor site. Genomic DNA for RPL22 from wild type and three mutant ery2 alleles was amplified and sequenced and revealed three different point mutations. Two alleles have premature stop codons and one allele changes a splice site acceptor site.

The clp1 Gene of the Mushroom Coprinus cinereus Is Essential for A-Regulated Sexual Development

Genetics ◽  
2001 ◽  
Vol 157 (1) ◽  
pp. 133-140
Author(s):  
Kazumi Inada ◽  
Yoshinori Morimoto ◽  
Toshihide Arima ◽  
Yukio Murata ◽  
Takashi Kamada
Keyword(s):  
Sexual Development ◽  
Genomic Dna ◽  
Mutant Allele ◽  
Coprinus Cinereus ◽  
Mating Partner ◽  
Essential Step ◽  
Genes Encoding ◽  
Dna Fragment ◽  
Necessary And Sufficient ◽  
Novel Protein

Abstract Sexual development in the mushroom Coprinus cinereus is under the control of the A and B mating-type loci, both of which must be different for a compatible, dikaryotic mycelium to form between two parents. The A genes, encoding proteins with homeodomain motifs, regulate conjugate division of the two nuclei from each mating partner and promote the formation of clamp connections. The latter are hyphal configurations required for the maintenance of the nuclear status in the dikaryotic phase of basidiomycetes. The B genes encode pheromones and pheromone receptors. They regulate the cellular fusions that complete clamp connections during growth, as well as the nuclear migration required for dikaryosis. The AmutBmut strain (326) of C. cinereus, in which both A- and B-regulated pathways are constitutively activated by mutations, produces, without mating, dikaryon-like, fertile hyphae with clamp connections. In this study we isolated and characterized clampless1-1 (clp1-1), a mutation that blocks clamp formation, an essential step in A-regulated sexual development, in the AmutBmut background. A genomic DNA fragment that rescues the clp1-1 mutation was identified by transformations. Sequencing of the genomic DNA, together with RACE experiments, identified an ORF interrupted by one intron, encoding a novel protein of 365 amino acids. The clp1-1 mutant allele carries a deletion of four nucleotides, which is predicted to cause elimination of codon 128 and frameshifts thereafter. The clp1 transcript was normally detected only in the presence of the A protein heterodimer formed when homokaryons with compatible A genes were mated. Forced expression of clp1 by promoter replacements induced clamp development without the need for a compatible A gene combination. These results indicate that expression of clp1 is necessary and sufficient for induction of the A-regulated pathway that leads to clamp development.

scholarly journals Selective enrichment of a large size genomic DNA fragment by affinity capture: an approach for genome mapping

1990 ◽  
Vol 18 (7) ◽  
pp. 1789-1795 ◽  
Author(s):  
Rajendra P. Kandpal ◽  
David C. Ward ◽  
Sherman M. Weissman
Keyword(s):  
Genomic Dna ◽  
Genome Mapping ◽  
Affinity Capture ◽  
Selective Enrichment ◽  
Large Size ◽  
Dna Fragment

scholarly journals Factor D̄ of the alternative pathway of human complement. Purification, alignment and N-terminal amino acid sequences of the major cyanogen bromide fragments, and localization of the serine residue at the active site

1980 ◽  
Vol 187 (3) ◽  
pp. 863-874 ◽  
Author(s):  
D M Johnson ◽  
J Gagnon ◽  
K B Reid
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Amino Acid Sequence ◽  
Alternative Pathway ◽  
Amino Acid Sequences ◽  
Serine Residue ◽  
Amino Acid Sequence Analysis ◽  
Terminal Amino Acid ◽  
Terminal Amino ◽  
Terminal Amino Acid Sequence

The serine esterase factor D of the complement system was purified from outdated human plasma with a yield of 20% of the initial haemolytic activity found in serum. This represented an approx. 60 000-fold purification. The final product was homogeneous as judged by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis (with an apparent mol.wt. of 24 000), its migration as a single component in a variety of fractionation procedures based on size and charge, and its N-terminal amino-acid-sequence analysis. The N-terminal amino acid sequence of the first 36 residues of the intact molecule was found to be homologous with the N-terminal amino acid sequences of the catalytic chains of other serine esterases. Factor D showed an especially strong homology (greater than 60% identity) with rat ‘group-specific protease’ [Woodbury, Katunuma, Kobayashi, Titani, & Neurath (1978) Biochemistry 17, 811-819] over the first 16 amino acid residues. This similarity is of interest since it is considered that both enzymes may be synthesized in their active, rather than zymogen, forms. The three major CNBr fragments of factor D, which had apparent mol.wts. of 15 800, 6600 and 1700, were purified and then aligned by N-terminal amino acid sequence analysis and amino acid analysis. By using factor D labelled with di-[1,3-14C]isopropylphosphofluoridate it was shown that the CNBr fragment of apparent mol.wt. 6600, which is located in the C-terminal region of factor D, contained the active serine residue. The amino acid sequence around this residue was determined.

scholarly journals One of the tightly clustered genes of the mouse surfeit locus is a highly expressed member of a multigene family whose other members are predominantly processed pseudogenes.

1988 ◽  
Vol 8 (9) ◽  
pp. 3898-3905 ◽  
Author(s):  
C Huxley ◽  
T Williams ◽  
M Fried
Keyword(s):  
Multigene Family ◽  
Open Reading Frame ◽  
The Other ◽  
Base Pairs ◽  
Regulation Of Expression ◽  
Reading Frame ◽  
Processed Pseudogenes ◽  
Dna Fragment ◽  
Basic Polypeptide

The mouse surfeit locus is unusual in that it contains a number of closely clustered genes (Surf-1, -2, and -4) that alternate in their direction of transcription (T. Williams, J. Yon, C. Huxley, and M. Fried, Proc. Natl. Acad. Sci. USA 85:3527-3530, 1988). The heterogeneous 5' ends of Surf-1 and Surf-2 are separated by 15 to 73 base pairs (bp), and the 3' ends of Surf-2 and Surf-4 overlap by 133 bp (T. Williams and M. Fried, Mol. Cell. Biol. 6:4558-4569, 1986; T. Williams and M. Fried, Nature (London) 322:275-279, 1986). A fourth gene in this locus, Surf-3, which is a member of a multigene family, has been identified. The poly(A) addition site of Surf-3 lies only 70 bp from the poly(A) addition site of Surf-1. Transcription of Surf-3 has been studied in the absence of the other members of its multigene family after transfection of a cloned genomic mouse DNA fragment, containing the Surf-3 gene, into heterologous monkey cells. Surf-3 specifies a highly expressed 1.0-kilobase mRNA that contains a long open reading frame of 266 amino acids, which would encode a highly basic polypeptide (23% Arg plus Lys). The other members of the Surf-3 multigene family are predominantly, if not entirely, intronless pseudogenes with the hallmarks of being generated by reverse transcription. The role of the very tight clustering on regulation of expression of the genes in the surfeit locus is discussed.

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