Isolation and characterization of alcohol dehydrogenase cDNAs from Pinusradiata

1990 ◽  
Vol 20 (9) ◽  
pp. 1343-1350 ◽  
Author(s):  
C. S. Kinlaw ◽  
D. E. Harry ◽  
R. R. Sederoff
Keyword(s):  
Alcohol Dehydrogenase ◽  
Southern Blot Analysis ◽  
Amino Acid Sequences ◽  
Coding Region ◽  
Coding Sequences ◽  
The Third ◽  
Isolation And Characterization ◽  
Adh Genes ◽  
Restriction Fragment Pattern

Three alcohol dehydrogenase (ADH) cDNAs were isolated from Pinusradiata. Two of the cDNAs appear to correspond to alleles of one ADH locus, and the third cDNA appears to correspond to a second ADH locus. Nucleotide and amino acid sequences of the coding region of ADH genes from the following species were compared: Pinusradiata, Zeamays, Hordeumvulgare, Triticumaestivum, Oryza sativa, Pisumsativum, and Arabidopsisthaliana. A phylogenetic tree was constructed of coding sequences of pine and angiosperm ADH genes. This tree shows three plant ADH clusters: monocot, dicot, and pine. The distance between pine and the two angiosperms is only slightly greater than the distance between either angiosperm, supporting the fossil evidence that suggests that monocots and dicots diverged from each other shortly after angiosperms diverged from gymnosperms. The structure of pine ADH genes was investigated by Southern blot analysis. The restriction fragment pattern of ADH genes from pines is more complex than the pattern from angiosperm genes, suggesting that pine ADH genes are either larger or more numerous than their angiosperm counterparts.

Amino acid sequence of penicillopepsin. III. Isolation and characterization of amino terminal, tryptic, and tryptophanyl peptides

1976 ◽  
Vol 54 (10) ◽  
pp. 895-901
Author(s):  
C. Ian Harris ◽  
Leticia Rao ◽  
Paul Shutsa ◽  
Alexander Kurosky ◽  
Theo Hofmann
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Cyanogen Bromide ◽  
Amino Acid Sequences ◽  
Affinity Column ◽  
Amino Terminal ◽  
Tryptic Digest ◽  
The Third ◽  
Isolation And Characterization

The amino acid sequences of peptides isolated from a tryptic digest of penicillopepsin (EC 3.4.23.7), a subtilisin (EC 3.4.21.14) digest of maleylated penicillopepsin, and a chymotryptic digest of penicillopepsin modified with dinitrophenylsulfenyl (DNPS) chloride have been determined. The first two digests identified four of the five lysyl residues of the enzyme as well as the N-terminal peptide. The third digest provided overlaps at three of the tryptophanyl residues. The DNPS-tryptophan peptides were isolated on an affinity column prepared by coupling dinitrophenyl antibody raised in sheep to cyanogen bromide-activated Sepharose.

Isolation and characterization of the pea cytochrome c oxidase Vb gene

Genome ◽  
2006 ◽  
Vol 49 (11) ◽  
pp. 1481-1489 ◽  
Author(s):  
Nakao Kubo ◽  
Shin-ichi Arimura ◽  
Nobuhiro Tsutsumi ◽  
Koh-ichi Kadowaki ◽  
Masashi Hirai
Keyword(s):  
Amino Acid ◽  
Cytochrome C ◽  
Cytochrome C Oxidase ◽  
Fluorescent Proteins ◽  
Amino Acid Sequences ◽  
Homology Search ◽  
Coding Region ◽  
Angiosperm Evolution ◽  
Isolation And Characterization ◽  
Duplication Events

Three copies of the gene that encodes cytochrome c oxidase subunit Vb were isolated from the pea (PscoxVb-1, PscoxVb-2, and PscoxVb-3). Northern Blot and reverse transcriptase-PCR analyses suggest that all 3 genes are transcribed in the pea. Each pea coxVb gene has an N-terminal extended sequence that can encode a mitochondrial targeting signal, called a presequence. The localization of green fluorescent proteins fused with the presequence strongly suggests the targeting of pea COXVb proteins to mitochondria. Each pea coxVb gene has 5 intron sites within the coding region. These are similar to Arabidopsis and rice, although the intron lengths vary greatly. A phylogenetic analysis of coxVb suggests the occurrence of gene duplication events during angiosperm evolution. In particular, 2 duplication events might have occurred in legumes, grasses, and Solanaceae. A comparison of amino acid sequences in COXVb or its counterpart shows the conservation of several amino acids within a zinc finger motif. Interestingly, a homology search analysis showed that bacterial protein COG4391 and a mitochondrial complex I 13 kDa subunit also have similar amino acid compositions around this motif. Such similarity might reflect evolutionary relationships among the 3 proteins.

Amino acid sequence of penicillopepsin. I. Isolation and characterization of the chymotryptic peptides

1976 ◽  
Vol 54 (10) ◽  
pp. 872-884 ◽  
Author(s):  
Alexander Kurosky ◽  
Theo Hofmann
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Amino Acid Sequences ◽  
Acid Protease ◽  
Terminal Region ◽  
Isolation And Characterization

The amino acid sequences of 48 peptides obtained from a chymotryptic digest of the mould acid protease, penicillopepsin (EC 3.4.23.7), have been determined. These peptides established the sequences of 26 unique fragments of up to 28 residues in length. The 28-residue fragment was identified as the N-terminal region. The C-terminal region is represented by a 13-residue fragment. The amino acids contained in these fragments account for some 85% of the residues of the enzyme.

Isolation and characterization of an expressed napin gene fromBrassica rapa

1991 ◽  
Vol 1 (4) ◽  
pp. 209-219 ◽  
Author(s):  
Jean C. Kridl ◽  
David W. McCarter ◽  
Ronald E. Rose ◽  
Donna E. Scherer ◽  
Deborah S. Knutzon ◽  
...  
Keyword(s):  
Brassica Rapa ◽  
Storage Protein ◽  
Protein Gene ◽  
Coding Region ◽  
Coding Regions ◽  
Endogenous Expression ◽  
Isolation And Characterization ◽  
Glucuronidase Reporter ◽  
Transgenic Rapeseed

AbstractAn expressed napin storage protein gene fromBrassica rapa, BcNA1, has been cloned and sequenced. The gene is a member of a family of four to seven napin genes inB. rapaand is highly expressed in developing seeds. An expression cassette containing the DNA flanking the napin coding region of BcNA1 has been engineered and demonstrated to function appropriately, as compared with the gene's endogenous expression, in transgenic rapeseed using the β-glucuronidase reporter gene. TheB. rapaBcNA1 gene and aB. napusnapin gene, gNa, share extremely high nucleotide homology not only throughout their coding regions, but over a DNA locus comprising 4.3 kb. We suggest the gNa gene was contributed by the originalB. rapaprogenitor of the amphidiploidB. napus.

Isolation and Characterization of Rhodopseudomonas sp. S9-1 Capable of Degrading Pyrazosulfuron-Ethyl

2011 ◽  
Vol 356-360 ◽  
pp. 1152-1163 ◽  
Author(s):  
Le Bin Yin ◽  
Yong Liu ◽  
De Yong Zhang ◽  
Song Bai Zhang
Keyword(s):  
Contaminated Soil ◽  
Acetolactate Synthase ◽  
Optimal Growth ◽  
Degradation Process ◽  
Amino Acid Sequences ◽  
Rrna Gene ◽  
Photosynthetic Membrane ◽  
Physiological And Biochemical Characteristics ◽  
Isolation And Characterization

A bacterial strain S9-1capable of degrading sulfonylurea herbicide pyrazosulfuron-ethyl (PSE) was isolated from contaminated soil through the enrichment incubation method. Based on morphology, colony and cultural properties, physiological and biochemical characteristics, living-cell absorption spectra, internal photosynthetic membrane, and phylogenetics of its 16S rRNA gene sequence, S9-1was preliminarily identified as belonging to the genus Rhodopseudomonas, a group of photosynthetic bacteria (PSB). The effects of PSE concentration, pH, and temperature on biodegradation were examined. The degradation rate was found to decrease with increasing PSE concentration. Optimal growth pH and temperature were found to be 7.0 and 30°C, respectively. The strain was able to degrade 47.51% of PSE at a concentration of 100 mg ml-1after 7 days of incubation at 30°C and could tolerate 800 mg ml-1PSE. S9-1was also able to completely co-metabolically transform 100 mg ml-1PSE at 30°C, pH 7.0, and 7500 lux in 15 days. As the concentration of PSE increased, the degradation process took longer to complete. The fragment encoding acetolactate synthase (ALS) gene from S9-1was cloned and sequenced. Comparison of deduced amino acid sequences was implemented, and the conserved sites were analyzed. To our knowledge, this is the first report of PSB in PSE biodegradation. These results highlight the potential of this bacterium as a detoxifying agent for use with PSE-contaminated soil and wastewater.

Characterization of the mouse thyrotrophin-releasing hormone receptor gene: an exon corresponds to a deletion in the rat cDNA

1993 ◽  
Vol 11 (2) ◽  
pp. 141-149 ◽  
Author(s):  
S M Duthie ◽  
P L Taylor ◽  
K A Eidne
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Stop Codon ◽  
Amino Acid Sequences ◽  
R Gene ◽  
Coding Region ◽  
Thyrotrophin Releasing Hormone ◽  
Exon 2 ◽  
Exon 4

ABSTRACT The cloning and characterization of the mouse TRH receptor (TRH-R) gene revealed an untranslated exon (exon 1), a single intron and an upstream dinucleotide repeat sequence (d(TG)16.d(AG)21) in the 5′ untranslated region (UTR). The coding region was contained almost entirely on a second exon (exon 2), with the final amino acid and stop codon at the COOH terminus of the gene encoded by a third exon (exon 3) flanked by two introns. The 3′ UTR was contained on the remainder of exon 3 and on the final exon (exon 4). Exon 3 (228 bp) corresponds exactly to a 228 bp deletion that exists in the rat TRH-R cDNA, but not in the mouse cDNA. The mouse TRH-R cDNA encodes a protein of 393 amino acids which is 96% homologous to the rat TRH-R protein of 412 amino acids, but is 19 amino acids shorter at its COOH terminus. The coding sequence for these 19 amino acids (plus 1 extra amino acid) does exist in the mouse TRH-R gene, but the sequence is encoded by exon 4, separated from the rest of the coding region by the stop codon and 223 bp of 3′ UTR on exon 3. Splicing of exon 3 in the mouse TRH-R gene would remove the last amino acid, the stop codon and the 223 bp of 3′ UTR, allowing transcription to continue into the 3′ UTR on exon 4, which encodes the 19 extra amino acids found in the rat cDNA. This would then result in an alternative 412 amino acid version of the mouse TRH-R protein, with 95% homology to the rat TRH-R. This study focused on the structural differences in the intracellular COOH-terminal tail of the receptor, which is known to be a functionally important domain in other members of the G protein-coupled receptor family. We have also recently characterized the human TRH-R cDNA, which revealed a third variant at the COOH terminus. Comparisons between mouse, rat and human TRH-Rs show that the amino acid sequences are virtually identical. However, significant differences between these species exist at the COOH terminus, with each TRH-R having a unique form of the COOH-terminal tail, beginning at exactly the same site and encoding 1, 20 and 6 amino acids in the mouse, rat and human respectively.

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