scholarly journals Identification and characterization of a resistance gene analog (RGA) from the Caricaceae Dumort family

2006 ◽  
Vol 28 (3) ◽  
pp. 458-462 ◽  
Author(s):  
Paulo de Paiva Rosa Amaral ◽  
Paulo César Martins Alves ◽  
Natália Florêncio Martins ◽  
Felipe Rodrigues da Silva ◽  
Guy de Capdeville ◽  
...  
Keyword(s):  
Carica Papaya ◽  
R Genes ◽  
Solanum Bulbocastanum ◽  
Conserved Motifs ◽  
Vector Sequences ◽  
High Degree ◽  
Identification And Characterization ◽  
Lrr Domain ◽  
Amino Acid Sequence Conservation

The majority of cloned resistance (R) genes characterized so far contain a nucleotide-binding site (NBS) and a leucine-rich repeat (LRR) domain, where highly conserved motifs are found. Resistance genes analogs (RGAs) are genetic markers obtained by a PCR-based strategy using degenerated oligonucleotide primers drawn from these highly conserved "motifs". This strategy has the advantage of the high degree of structural and amino acid sequence conservation that is observed in R genes. The objective of the present study was to search for RGAs in Carica papaya L. and Vasconcellea cauliflora Jacq. A. DC. Out of three combinations of primers tested, only one resulted in amplification. The amplified product was cloned in pCR2.1TOPO and than sequenced using M13 forward and reverse primers. Forty-eight clones were sequenced from each species. The 96 sequences generated for each species were cleaned of vector sequences and clustered using CAP3 assembler. From the GENEBANK, one RGA was identified in C. papaya showing a BlastX e-value of 2x10-61 to the gb|AAP45165.1| putative disease resistant protein RGA3 (Solanum bulbocastanum). To the extent of our knowledge this is the first report of a RGA in the Caricaceae Dumort family. Preliminary structural studies were performed to further characterize this putative NBS-LRR type protein. Efforts to search for other RGAs in papaya should continue, mostly to provide basis for the development of transgenic papaya with resistance to diseases.

scholarly journals Plasmid-mediated trimethoprim-resistance in Staphylococcus aureus. Characterization of the first gram-positive plasmid dihydrofolate reductase (type S1)

1987 ◽  
Vol 243 (1) ◽  
pp. 309-312 ◽  
Author(s):  
H K Young ◽  
R A Skurray ◽  
S G B Amyes
Keyword(s):  
Staphylococcus Aureus ◽  
Dihydrofolate Reductase ◽  
Specific Activity ◽  
Gram Positive ◽  
Heat Stable ◽  
Dihydrofolate Reductases ◽  
High Degree ◽  
Moderately Resistant ◽  
Identification And Characterization

The trimethoprim-resistance gene located on plasmid pSK1, originally identified in a multi-resistant Staphylococcus aureus from Australia, encodes the production of a dihydrofolate reductase (type S1), which confers a high degree of resistance to its host and is quite unlike any plasmid-encoded dihydrofolate reductase hitherto described. It has a low Mr (19,700) and has a higher specific activity than the constitutive Gram-negative plasmid dihydrofolate reductases. The type S1 enzyme is heat-stable and has a relatively low affinity for the substrate, dihydrofolate (Km 10.8 microM). It is moderately resistant to trimethoprim, and is competitively inhibited by this drug with an inhibitor-binding constant of 11.6 microM. This is the first identification and characterization of a plasmid-encoded trimethoprim-resistant dihydrofolate reductase derived from a Gram-positive species.

scholarly journals Antimicrobial Spectrum, Growth/Killing Kinetics, Conventional/Molecular Assay of Characterizing Non-Leguminous Endophytic Bacteria and Fungi from Helianthus annuus, Carica papaya and Lycoperesicum solanum

2021 ◽  
Vol 2 (10) ◽  
pp. 1018-1034
Author(s):  
Osuntokun OT ◽  
Azuh VO ◽  
Adejoro BF ◽  
Akele EO
Keyword(s):  
Helianthus Annuus ◽  
Death Rate ◽  
Endophytic Bacteria ◽  
Carica Papaya ◽  
Molecular Assay ◽  
Molecular Method ◽  
Preliminary Identification ◽  
Bacteria And Fungi ◽  
Identification And Characterization

The aim of this study is to comparative study between conventional and molecular assay of isolation, identification and characterization of non-leguminous endophytic bacteria and fungi in the leguminous root samples. The plant root samples, Helianthus annuus, Carica papaya and Lycoperesicum solanum (Sunflower root and stem, pawpaw root and stem, and tomato root and stem from Adekunle Ajasin University School farm, Akungba Akoko, Ondo state, Nigeria. The isolation of endophytic bacteria were performed using the conventional method of isolation (biochemical test) and characterization were done using both the conventional and molecular method of bacteria characterization. The antibiotic susceptibility test (Antibiogram) was observed using disc diffusion. The four bacteria identified were Bacillus cereus, Enterobacter sp. Actnomycoses sp. and Aeromonas sp. for conventional method and Fusarium solani, Fusarium vortecelium and Bacillus thuringiensis for molecular method as confirmatory point of view. In this study, all isolated organisms tends to be Gram positive using the gram staining technique. Antibiogram shows the zones of inhibition with diameter ranging from 0-20 mm, Enterobacter sp. were more sensitive to the various antibiotics used. Ultraviolet spectrophotometer was also used to determine the growth dynamic as well as the death rate of the isolates, the addition of antibiotics (ciprofloxacin) to the isolates at the 24th hour speed up the death rate of the isolates from non-leguminous endophytic bacteria. After the preliminary identification of the bacteria isolates and the confirmatory identification of both bacteria and fungi isolates of the non-leguminous endophytic microorganism, it was noted that the preliminary identification was only able to achieve the genus level of taxonomic characterization, While the molecular method confirm the molecular sub level identification of isolates depletes the absolute taxonomic identification and characterization to the sub-species level. The results of this study validates the use of molecular sequencing for the assay identification and characterization of non-leguminous endophytic bacteria and fungi as the easy and best mode of identification of both bacteria and fungi isolates as a veritable tools for research purposes.

scholarly journals In-Silico Characterization and Expression Analysis of NB-ARC Genes in Response to Erwinia mallotivora in Carica papaya

2021 ◽  
Vol 50 (9) ◽  
pp. 2591-2602
Author(s):  
Nur Syazana Abu Bakar ◽  
Noor Baity Saidi ◽  
Lina Rozano ◽  
Mohd Puad Abdullah ◽  
Suhaina Supian
Keyword(s):  
Disease Resistance ◽  
Plant Disease Resistance ◽  
Carica Papaya ◽  
R Genes ◽  
Binding Surface ◽  
Transcriptome Database ◽  
Defence Signalling ◽  
In Silico Characterization ◽  
Lrr Domain

Disease resistance in plants is commonly associated with resistance (R) genes that encode nucleotide binding site-leucine rich repeat (NBS-LRR) domains that are essential for pathogen recognition and defence signalling. In this study, we identified and analyzed the sequence of putative pathogen-responsive NB-ARC transcripts from Carica papaya transcriptome database, carried out the structural and phylogenetic analysis, and determined the expression profile of the transcripts in C. papaya challenged with Erwinia mallotivora. The findings indicate CpNBS1, the only pathogen-responsive NB-ARC protein identified in this study belongs to the CC-NBS-LRR group. Semi-quantitative PCR showed CpNBS1 was differentially expressed in response to E. mallotivora. Structural analysis of the 4993-Eksotika and 4993-Viorica translated proteins showed striking differences in terms of the number of β-sheets and α-helixes as well their ligand-binding surface, suggesting the role of the LRR domain in determining the specificity of recognition of E. mallotivora effector. Collectively, this study provides new insights into the role of NBS-LRR genes in C. papaya and its implications for enhancing of plant disease resistance through genetic engineering.

Identification and characterization of chymopapains A and B from dried and fresh latex of Carica papaya

1986 ◽  
Vol 14 (6) ◽  
pp. 1226-1227 ◽  
Author(s):  
BALDEV S. BAINES ◽  
KEITH BROCKLEHURST ◽  
RAYMOND McKEE ◽  
MAIREAD O'DRISCOLL ◽  
ERDJAN SALIH ◽  
...  
Keyword(s):  
Carica Papaya ◽  
Identification And Characterization

scholarly journals Rhesus Monkey (Macaca mulatta) Mucosal Antimicrobial Peptides Are Close Homologues of Human Molecules

2001 ◽  
Vol 8 (2) ◽  
pp. 370-375 ◽  
Author(s):  
Robert Bals ◽  
Christiane Lang ◽  
Daniel J. Weiner ◽  
Claus Vogelmeier ◽  
Ulrich Welsch ◽  
...  
Keyword(s):  
Rhesus Monkey ◽  
Antimicrobial Peptides ◽  
Polyclonal Antibodies ◽  
Amino Acid Sequences ◽  
Model Organisms ◽  
Immune Functions ◽  
Mucosal Surfaces ◽  
High Degree ◽  
Identification And Characterization

ABSTRACT One component of host defense at mucosal surfaces appears to be epithelium-derived antimicrobial peptides. Molecules of the defensin and cathelicidin families have been studied in several species, including human and mouse. We describe in this report the identification and characterization of rhesus monkey homologues of human mucosal antimicrobial peptides. Using reverse transcriptase PCR methodology, we cloned the cDNAs of rhesus monkey β-defensin 1 and 2 (rhBD-1 and rhBD-2) and rhesus monkey LL-37/CAP-18 (rhLL-37/rhCAP-18). The predicted amino acid sequences showed a high degree of homology to the human molecules. The expression of the monkey antimicrobial peptides was analyzed using immunohistochemistry with three polyclonal antibodies to the human molecules. As in humans, rhesus monkey antimicrobial peptides are expressed in epithelia of various organs. The present study demonstrates that β-defensins and cathelicidins of rhesus monkeys are close homologues to the human molecules and indicate that nonhuman primates represent valid model organisms to study innate immune functions.

scholarly journals Identification and characterization of Saccharomyces cerevisiae yapsin 3, a new member of the yapsin family of aspartic proteases encoded by the YPS3 gene

1999 ◽  
Vol 339 (2) ◽  
pp. 407-411 ◽  
Author(s):  
Vicki OLSEN ◽  
Niamh X. CAWLEY ◽  
Jakob BRANDT ◽  
Michi EGEL-MITANI ◽  
Y. Peng LOH
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Amino Acid ◽  
Cleavage Sites ◽  
Basic Residue ◽  
Aspartic Proteases ◽  
N Terminus ◽  
High Degree ◽  
Identification And Characterization ◽  
Degree Of Similarity

A new aspartic protease from Saccharomyces cerevisiae, with a high degree of similarity with yapsin 1 and yapsin 2 and a specificity for basic residue cleavage sites of prohormones, has been cloned. This enzyme was named yapsin 3. Expression of a C-terminally truncated non-membrane anchored yapsin 3 in yeast yielded a heterogeneous protein between 135–200 kDa which, upon treatment with endoglycosidase H, migrated as a 60 kDa form. Amino-acid analysis of the N-terminus of expressed yapsin 3 revealed two different N-terminal residues, serine-48 and phenylalanine-54, which followed a dibasic and a monobasic residue respectively. Cleavage of several prohormones by non-anchored yapsin 3 revealed a specificity distinct from that of yapsin 1.

scholarly journals Morphological characterization of selected papaya (Carica papaya L.) inbreds and hybrids

2009 ◽  
pp. 102-112
Author(s):  
Florence Lasalita-Zapico ◽  
Violeta Villegas
Keyword(s):  
Carica Papaya ◽  
Genetic Relatedness ◽  
Morphological Characteristics ◽  
Morphological Characterization ◽  
Accurate Estimation ◽  
Phenotypic Traits ◽  
Complete Dominance ◽  
Phylogenetic Affinities ◽  
High Degree

Morphological analysis was undertaken to assess the degree of genetic relatedness and to characterize selected papaya (Carica papaya L.) inbreds and hybrids. Transmission of phenotypic traits from inbred parents to hybrid progeny followed the Mendelian pattern (complete dominance). The clustering mechanism separated the papaya genotypes into two groups. It was also revealed that some inbred lines presumably of very diverse origins exhibited similar morphological characteristics, raising the possibility that they have phylogenetic affinities and/or common origins. Screening for morphological traits with a high degree of polymorphism and with invariable expressions of the phenotypes would help in the identification of markers for hybrid identification and also in the accurate estimation of genetic relatedness among these hybrids and their parents.

scholarly journals Identification and characterization of REC66, a Ty1-copia-like retrotransposon in the genome of red flower of Mirabilis jalapa L.

2017 ◽  
Vol 69 (2) ◽  
pp. 315-322
Author(s):  
Jiang Shunri ◽  
Liang Joshua ◽  
Feng Haiyan ◽  
Luo Dan ◽  
Blake Shester ◽  
...  
Keyword(s):  
Genetic Research ◽  
Plant Genome ◽  
Degenerate Primers ◽  
Genetic Studies ◽  
Gag Protein ◽  
Mirabilis Jalapa ◽  
High Degree ◽  
Plant Genome Evolution ◽  
Identification And Characterization

Mirabilis jalapa Lis the most commonly grown ornamental species of Mirabilis and is available in a range of brilliant colors. However, genetic research on Mirabilis jalapa Lis limited. Using fluorescent differential display (FDD) screening, we report the identification of a novel Ty1-copia-like retrotransposon in the genome of the red flower of Mirabilis jalapa L, and we named it REC66based on its sequence homology to the GAG protein from Ty1-copiaretrotransposon. Using degenerate primers based on the DNA sequence of REC66, a total of fourteen different variants in reverse transcriptase (RT) sequence were recovered from the genomic DNA. These RT sequences show a high degree of heterogeneity characterized mainly by deletion mutation; they can be divided into three subfamilies, of which the majority encode defective RT. This is the first report of a Ty1-copiaretrotransposon in Mirabilis jalapa L. The finding could be helpful for the development of new molecular markers for genetic studies, particularly on the origin and evolutionary relationships of M. jalapa L, and the study of Ty1-copiaretrotransposons and plant genome evolution in the genus Mirabilisor family Nyctaginaceae.

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