POLIMORFISME GEN NATURAL RESISTANCE-ASSOCIATED MACROPHAGE PROTEIN-1 (NRAMP-1) DENGAN PCR-RFLP DARI EKSTRAK SALIVA MENGGUNAKAN ENZIM RESTRIKSI AVA II

Telah dilakukan penelitian mengenai Polimorfisme Gen Natural Resistance-Associated Macrophage Protein-1 (NRAMP-1) Dengan PCR-RFLP Dari Ekstrak Saliva Menggunakan Enzim Restriksi Ava II. Penelitian ini dilakukan di Laboratorium Biologi Molekuler dan imunologi Bagian Mikrobiologi, Fakultas Kedokteran, Universitas Hasanuddin, Makassar. Penelitian ini bertujuan untuk mengetahui polimorfisme pada gen Natural resistance-associated macrophage protein-1 (Nramp-1) khususnya pada lokus D543N dari hasil ekstraksi DNA saliva dengan enzim restriksi Ava II. Sampel saliva diekstraksi dengan metode Boom sehingga diperoleh DNA genom, selanjutnya diamplifikasi dengan menggunakan enzim Ava II. Hasil Elektroforesis gel yang dilakukan menunjukkan tidak terdapat perbedaan pola pemotongan pita DNA. Sampel saliva dari 10 responden (100%) tidak terjadi pemotongan. Hasil ini menunjukkan bahwa tidak terdapat perbedaan polimorfisme dari saliva responden

Mycobacterium tuberculosis Expresses a Novel Ph-Dependent Divalent Cation Transporter Belonging to the Nramp Family

Mammalian natural resistance–associated macrophage protein (Nramp) homologues are important determinants of susceptibility to infection by diverse intracellular pathogens including mycobacteria. Eukaryotic Nramp homologues transport divalent cations such as Fe2+, Mn2+, Zn2+, and Cu2+. Mycobacterium tuberculosis and Mycobacterium bovis (bacillus Calmette-Guérin [BCG]) also encode an Nramp homologue (Mramp)

Analysis of genetic variation of inducible nitric oxide synthase and natural resistance-associated macrophage protein 1 loci in Malaysian native chickens

The genetic diversity of 100 Malaysian native chickens was investigated using polymerase chain reaction-restriction fragment polymorphism (PCR-RFLP) for two candidate genes: inducible nitric oxide synthase (INOS) and natural resistance-associated macrophage protein 1 (NRAMP1). The two genes were selected because of their important role in chicken’s immune system. INOS and NRAMP1 PCR products were digested by AluI and SacI restriction enzymes, respectively. The restriction digests produced fragment sizes of 322 and 173 bp for INOS and 722 and 79 bp for NRAMP1 as one allele and an undigested PCR product as the other allele. Both loci were polymorph, however only INOS gene showed Hardy-Weinberg equilibrium. Average heterozygosity and the Shannon information index (I) was 0.43 and 0.62 for INOS and 0.48 and 0.68 for NRAMP1 genes, respectively. The observed polymorphism in this study shows the ability of these candidate genes in marker assisted selection and introgression programs to increase resistance to diseases in both Malaysian native and commercial chickens.Key words: Malaysian native chickens, polymorphism, inducible nitric oxide synthase (INOS), natural resistance-associated macrophage protein 1 (NRAMP1)

In Vitro Modulation Of Gene Expression In Bovine Blood Neutrophils

Concerns for animal welfare, food safety, and security fuel interest in understanding modulators of innate immunity. The accumulation of neutrophils and activation of toll-like receptors (TLRs) at infection sites are critical events of innate host defense. The objectives of this study were to evaluate the effects of Lipopolysaccaride (LPS), Peptidoglycan (PGN), Nystatin (NYS), and Quebracho (Q) extract on bovine neutrophils activation in relation to expression of genes encoding TLR2, TLR4, natural resistance-associated macrophage protein 1 (Nramp1), and the cytokines TNF-α and IL-10

Polimorfismos na região 5' UTR do gene NRAMP1 em equinos da raça pantaneira.

O gene NRAMP1 (Natural Resistance Associated Macrophage Protein 1) foi identificado em várias espécies animais e está relacionado à resistência natural a patógenos intracelulares. Este trabalho teve por objetivo avaliar, via sequenciamento, polimorfismos existentes na região 5' não traduzida (UTR) deste gene

Natural Resistance-Associated Macrophage Protein Is a Cellular Receptor for Sindbis Virus in Both Insect and Mammalian Hosts

SummaryAlphaviruses, including several emerging human pathogens, are a large family of mosquito-borne viruses with Sindbis virus being a prototypical member of the genus. The host factor requirements and receptors for entry of this class of viruses remain obscure. Using a Drosophila system, we identified the divalent metal ion transporter natural resistance-associated macrophage protein (NRAMP) as a host cell surface molecule required for Sindbis virus binding and entry into Drosophila cells. Consequently, flies mutant for dNRAMP were protected from virus infection. NRAMP2, the ubiquitously expressed vertebrate homolog, mediated binding and infection of Sindbis virus into mammalian cells, and murine cells deficient for NRAMP2 were nonpermissive to infection. Alphavirus glycoprotein chimeras demonstrated that the requirement for NRAMP2 is at the level of Sindbis virus entry. Given the conserved structure of alphavirus glycoproteins, and the widespread use of transporters for viral entry, other alphaviruses may use conserved multipass membrane proteins for infection

Molecular characterization of the NRAMP1 gene in buffalo

NRAMP1 (natural-resistance-associated macrophage protein) gene influences the initial phase of bacterial cellular infections, regulating macrophage activation. Recent literature on buffalo has attempted to associate the genotypes at the polymorphic microsatellite, that is located in the 3'-UTR of the gene, with either susceptibility to brucellosis or improved macrophage function. However, contradictory results were reported. In the present work, we have sequenced the whole coding region, as well as part of the introns and UTRs, of the NRAMP1 gene in 49 Mediterranean buffaloes, including both serologically positive and negative animals to Brucella abortus test. We have detected 12 mutations. Nineteen haplotypes were built from the detected variant alleles, so demonstrating the high variability of this gene in buffalo, but no significant differences in haplotype frequencies were found between serologically positive/negative animals

Natural resistance-associated macrophage protein 1 gene polymorphisms in thalassemia patients with tuberculosis infection

that needs regular blood transfusions leading to accumulation of iron in the cells. This iron overload level in macrophage might cause intracellular bacteria, particularly Mycobacterium tuberculosis (MTB) to multiply. Polymorphisms in natural resistance-associated macrophage protein 1 (NRAMP1), a metal transporter across the phagosome membrane, play important role in regulating iron, which is also needed by MTB. Increased iron in thalassemia patients may have an increased potential risk for TB. Objective To compare natural resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms (INT4, D543N, and 3’UTR) in thalassemia patients with and without tuberculosis (TB) infection. Methods A cross-sectional measurement of NRAMP1 genetic polymorphisms was performed in pediatric thalassemia patients with TB (n=40) and without TB (n=50). Iron status including serum iron, total iron-binding capacity (TIBC), and ferritin, was compared between the two groups. The NRAMP1 genetic polymorphisms were analysed using polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP). Allelic and genotypic distributions of each polymorphism were assessed for possible associations with TB infection. Results Mean serum iron and TIBC in thalassemia patients with TB were higher compared to thalassemia patients without TB (mean serum: 166.26 vs. 134.92 μmol/L, respectively; P=0.026) and (mean TIBC: 236.78 vs. 195.84 μmol/L, respectively; P=0.029). In thalassemia patients with TB, we observed significantly higher frequency of the C allele in INT4 (10% vs. 2%, respectively; OR=5.44; 95%CI 1.1 to 26.4; P=0.02) and the TGTG deletion allele (78.8% vs. 51%, respectively; OR=3.56; 95%CI 1.83 to 6.9; P=0.0002) in 3’UTR polymorphisms than in thalassemia patients without TB. There were no significant  differences in distributions of the A allele between TB and non-TB groups (16.3% vs. 15%, respectively; P=0.84) or the GA genotype (32.5% vs. 30%, respectively; P=0.79) in D543N. Conclusion The NRAMP1 polymorphisms are known to be associated with major gene susceptibility to TB, and in our thalassemia patients this association is even more pronounced