Development of Brucellosis Vaccine Based on Determinant Antigenic of Outer Membrane Protein (OMP) 36 kDa From Brucella abortus Local Isolate

2017 ◽  
Vol 9 (3) ◽  
Author(s):  
Aulanni’am Aulanni’am ◽  
Wiwiek Tyasningsih ◽  
Dyah Kinasih Wuragil ◽  
Fedik Abdul Rantam
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Membrane Protein ◽  
Outer Membrane Protein ◽  
Brucella Abortus ◽  
Epitope Prediction ◽  
Mhc I ◽  
Mhc Ii ◽  
The Past ◽  
Local Isolate

Brucellosis is a disease that can be prevented through vaccination. Yet, the effectiveness of the vaccination to fight this disease is considered weak. Fortunately, attempts to modify brucellosis vaccine is still keep going. Some brucellosis vaccines have been found and developed in the past time such as the vaccine B.abortus strain 19-BA and 104M which was made from weakened microbes which had been widely used in Uni Soviet and China. The other brucellosis vaccine that were used in the past were the phenolinsoluble peptidoglycan vaccine which was made in France and polysaccharideprotein vaccine which was used in Russia. This research attempted to see the determinant of antigenic Outer Membrane Protein (OM) 36 kDa Brucella abortus local isolation which has immunogenic character to be developed as an advanced brucellosis vaccine. The method used in this research was the Omp2 gene of Brucella abortus of local isolate employed the PCR technique. The result of the PCR was then sequenced to analyze the determinant antigenic and the bounding prediction of either the T cell or the B cell which were responsible for immune response. The result of this study showed that the gen Omp2 which encoded the OMP 36 kDa Brucella abortus of local isolation with primary JPF 5’ GCG CTC AGG CTG CCG ACG CAA 3’ and JPR 5’ CAT TGC GGT CGG TAC CGG AG 3’ targeted the gene 162 bp, was then translated into amino acids to be later undergo the in silico test using Kolaskar and Tongaonkar Antigenicity Prediction method. The epitope prediction resulted were MSRVCDAYGAGYFYI and TETCLRVHGYVRYD. The result of the epitope prediction of MSRVCDAYGAGYFYI showed that there was a bond with MHC I in YGAGYFYI of the 8th amino acid series to the 15th series, while the epitope prediction of TETCLRVHGYVRYD showed that there was a bond to the ETCLRVHGY of the series of amino acids number 2 to 10. Bond with MHC II existed in the amino acid series of MSRVCDAYGAGYFYI, while the bond with the B cells existed in BCSAYGA and CLRVHG amino acid series. This research has been successful in predicting the epitope of the OMP 36 kDa Brucella abortus of local isolate which had immunogenic characteristic for its ability to bond with the MHC I, MHC II and B cells.

scholarly journals Sequence Polymorphism, Predicted Secondary Structures, and Surface-Exposed Conformational Epitopes of Campylobacter Major Outer Membrane Protein

2000 ◽  
Vol 68 (10) ◽  
pp. 5679-5689 ◽  
Author(s):  
Qijing Zhang ◽  
Jerrel C. Meitzler ◽  
Shouxiong Huang ◽  
Teresa Morishita
Keyword(s):  
Amino Acid ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Outer Membrane Proteins ◽  
Major Outer Membrane Protein ◽  
Amino Acid Sequences ◽  
Variable Regions ◽  
Conserved Sequences ◽  
Conformational Epitopes

ABSTRACT The major outer membrane protein (MOMP), a putative porin and a multifunction surface protein of Campylobacter jejuni, may play an important role in the adaptation of the organism to various host environments. To begin to dissect the biological functions and antigenic features of this protein, the gene (designatedcmp) encoding MOMP was identified and characterized from 22 strains of C. jejuni and one strain of C. coli. It was shown that the single-copy cmp locus encoded a protein with characteristics of bacterial outer membrane proteins. Prediction from deduced amino acid sequences suggested that each MOMP subunit consisted of 18 β-strands connected by short periplasmic turns and long irregular external loops. Alignment of the amino acid sequences of MOMP from different strains indicated that there were seven localized variable regions dispersed among highly conserved sequences. The variable regions were located in the putative external loop structures, while the predicted β-strands were formed by conserved sequences. The sequence homology of cmp appeared to reflect the phylogenetic proximity of C. jejuni strains, since strains with identical cmp sequences had indistinguishable or closely related macrorestriction fragment patterns. Using recombinant MOMP and antibodies recognizing linear or conformational epitopes of the protein, it was demonstrated that the surface-exposed epitopes of MOMP were predominantly conformational in nature. These findings are instrumental in the design of MOMP-based diagnostic tools and vaccines.

Nutrition of the bacon pig. XIX. The requirement of the bacon pig for certain essential amino acids

1958 ◽  
Vol 50 (2) ◽  
pp. 230-242 ◽  
Author(s):  
R. E. Evans
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Energy Supply ◽  
Essential Amino Acids ◽  
Food Utilization ◽  
Biological Value ◽  
Potential Value ◽  
The Past

The addition of essential amino acids to all-vegetable diets, so as to raise the biological value of their proteins to the level of animal-protein concentrates, has often been advocated in the past by animal nutritionists. This aim was frustrated in practice since it was impossible, until recently, to secure the necessary supplies at prices that would make such additions economical. Feeding stuffs with added amino acids are now being sold commercially. This is possible following the manufacture of synthetic DL-methionine and crude L-lysine monochloride.It seemed desirable, therefore, to carry out investigations into the potential value of these amino acids to the pig feeder. This paper deals with the effect on growth, food utilization and retention of nitrogen of adding small amounts of lysine and methionine to the diet. An attempt is made to correlate the amino acid composition of the pig's diet with its rate of growth, the energy supply being adequate.

scholarly journals Characterization of Specific Immune Responses of Mice Inoculated with Recombinant Vaccinia Virus Expressing an 18-Kilodalton Outer Membrane Protein of Brucella abortus

2000 ◽  
Vol 7 (1) ◽  
pp. 114-118 ◽  
Author(s):  
Ramesh Vemulapalli ◽  
Silvio Cravero ◽  
Christine L. Calvert ◽  
Thomas E. Toth ◽  
Nammalwar Sriranganathan ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Vaccinia Virus ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Brucella Abortus ◽  
Virulent Strain ◽  
Shuttle Vector ◽  
Protein Fusion

ABSTRACT Using the shuttle vector pMCO2 and the vaccinia virus wild-type WR strain, we constructed a recombinant virus expressing an 18-kDa outer membrane protein of Brucella abortus. BALB/c mice inoculated with this virus produced 18-kDa protein-specific antibodies, mostly of immunoglobulin G2a isotype, and in vitro stimulation of splenocytes from these mice with purified maltose binding protein–18-kDa protein fusion resulted in lymphocyte proliferation and gamma interferon production. However, these mice were not protected against a challenge with the virulent strain B. abortus2308. Disruption of the 18-kDa protein's gene in vaccine strainB. abortus RB51 did not affect either the strain's protective capabilities or its in vivo attenuation characteristics. These observations suggest that the 18-kDa protein plays no role in protective immunity.

scholarly journals Molecular Cloning of the Pasteurella haemolytica pomA Gene and Identification of Bovine Antibodies against PomA Surface Domains

1999 ◽  
Vol 67 (9) ◽  
pp. 4968-4973 ◽  
Author(s):  
Hui Zeng ◽  
Karamjeet Pandher ◽  
George L. Murphy
Keyword(s):  
Amino Acid ◽  
Nucleotide Sequence ◽  
Membrane Protein ◽  
Amino Acid Sequence ◽  
Outer Membrane Protein ◽  
Immunoglobulin G ◽  
Pasteurella Haemolytica ◽  
Respiratory Pathogen ◽  
Surface Domains ◽  
Bovine Immunoglobulin

ABSTRACT The gene (pomA) encoding PomA, an OmpA-like major outer membrane protein of the bovine respiratory pathogen Pasteurella haemolytica, was cloned, and its nucleotide sequence was determined. The deduced amino acid sequence of PomA has significant identity with the sequences of other OmpA family proteins. Absorption of three different bovine immune sera with whole P. haemolytica cells resulted in a reduction of bovine immunoglobulin G reactivity with recombinant PomA in Western immunoblots, suggesting the presence of antibodies against PomA surface domains.

scholarly journals Cloning of the Gene Encoding the Actinobacillus actinomycetemcomitans Serotype b OmpA-Like Outer Membrane Protein

1999 ◽  
Vol 67 (2) ◽  
pp. 942-945 ◽  
Author(s):  
Hitoshi Komatsuzawa ◽  
Toshihisa Kawai ◽  
Mark E. Wilson ◽  
Martin A. Taubman ◽  
Motoyuki Sugai ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Amino Acid ◽  
Membrane Protein ◽  
Amino Acid Sequence ◽  
Molecular Mass ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Actinobacillus Actinomycetemcomitans ◽  
Gene Encoding ◽  
Serotype B

ABSTRACT The gene encoding an outer membrane protein A (OmpA)-like, heat-modifiable Omp of Actinobacillus actinomycetemcomitans ATCC 43718 (strain Y4, serotype b) was cloned by a PCR cloning procedure. DNA sequence analysis revealed that the gene encodes a protein of 346 amino acid residues with a molecular mass of 36.9 kDa. The protein expressed by the cloned gene reacted with a monoclonal antibody to the previously described 29-kDa Omp (Omp29) of strain Y4. This monoclonal antibody reacted specifically with Omp29 of A. actinomycetemcomitans (serotype b), but not with any Omp of Escherichia coli, including OmpA. This protein exhibited characteristic heat modifiability on sodium dodecyl sulfate-polyacrylamide gels, showing an apparent molecular mass of 29 kDa when unheated and a mass of 34 kDa when heated. The N-terminal amino acid sequence of the protein expressed inE. coli perfectly matched those deduced from the purified Omp29 of strain Y4. The deduced amino acid sequence of the gene coding for Omp29 from serotype b matched completely (except for valine at position 321) that of a recently reported omp34gene described for A. actinomycetemcomitans serotype c (NCTC 9710). Because of the conserved nature of the gene within these serotypes, we designated the gene described herein from serotype b asomp34.

scholarly journals High-resolution mapping of serovar-specific and common antigenic determinants of the major outer membrane protein of Chlamydia trachomatis.

1988 ◽  
Vol 167 (3) ◽  
pp. 817-831 ◽  
Author(s):  
R S Stephens ◽  
E A Wagar ◽  
G K Schoolnik
Keyword(s):  
Amino Acid ◽  
Chlamydia Trachomatis ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Antigenic Determinants ◽  
Amino Acid Peptide ◽  
Antigenic Domains ◽  
Species Specific ◽  
Specific Determinant

The principal surface protein antigen of Chlamydia trachomatis is the major outer membrane protein (MOMP). The MOMP is antigenically complex. Among the 15 serovars of C. trachomatis, mAbs define serovar-, subspecies-, and species-specific determinants on MOMP. The molecular basis of the antigenic diversity of these proteins is reflected in amino acid variable sequence domains. We have mapped the dominant topographic antigenic determinants of MOMP that are defined by mAbs. Using recombinant DNA approaches we have identified the linear distribution of two antigenic domains. One domain contains a serovar-specific determinant and the other contains subspecies- and species-specific determinants. These antigenic domains correspond to two amino acid sequence variable domains. Synthetic peptides were immunogenic and these resolved the serovar-specific determinant within a 14-amino acid peptide. The subspecies- and species-specific determinants were overlapping within a 16-amino acid peptide.

scholarly journals Novel Genetic and Phenotypic Heterogeneity in Bordetella bronchiseptica Pertactin

2001 ◽  
Vol 69 (3) ◽  
pp. 1917-1921 ◽  
Author(s):  
Karen B. Register
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Dna Sequence ◽  
Outer Membrane Protein ◽  
Phenotypic Heterogeneity ◽  
Bordetella Bronchiseptica ◽  
Amino Acid Repeat ◽  
Sequence Heterogeneity

ABSTRACT The Bordetella bronchiseptica outer membrane protein pertactin is believed to function as an adhesin and is an important protective immunogen. Previous sequence analysis of the pertactin gene identified two regions predicted to encode amino acid repeat motifs. Recent studies have documented DNA sequence heterogeneity in both regions. The present study describes additional variants in these regions, which form the basis for six novel pertactin types. Immunoblotting demonstrated phenotypic heterogeneity in pertactin consistent with the predicted combined sizes of the repeat regions. A revised system for classifying B. bronchiseptica pertactin variants is proposed.

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