• Asian-Australasian Journal of Animal Sciences
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• 제27권8호
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• pp.1189-1195
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• 2014

Natural resistance-associated macrophage protein 1 encoding gene (NRAMP1) plays an important role in immune response against intracellular pathogens. To evaluate the effects of NRAMP1 gene on immune capacity in pigs, tissue expression of NRAMP1 mRNA was observed by real time quantitative polymerase chain reaction (PCR), and the results revealed NRAMP1 expressed widely in nine tissues. One single nucleotide polymorphism (SNP) (ENSSSCG00000025058: g.130 C>T) in exon1 and one SNP (ENSSSCG00000025058: g.657 A>G) in intron1 region of porcine NRAMP1 gene were demonstrated by DNA sequencing and PCR-RFLP analysis. A further analysis of SNP genotypes associated with immune traits including contain of white blood cell (WBC), granulocyte, lymphocyte, monocyte (MO), rate of cytotoxin in monocyte (MC) and $CD4^-CD8^+$ T lymphocyte subpopulations in blood was carried out in four pig populations including Large White and three Chinese indigenous breeds (Wannan Black, Huai pig and Wei pig). The results showed that the SNP (ENSSSCG00000025058: g.130 C>T) was significantly associated with level of WBC % (p = 0.031), MO% (p = 0.024), MC% (p = 0.013) and $CD4^-CD8^+$ T lymphocyte (p = 0.023). The other SNP (ENSSSCG00000025058: g.657 A>G) was significantly associated with the level of MO% (p = 0.012), MC% (p = 0.019) and $CD4^-CD8^+$ T lymphocyte (p = 0.037). These results indicate that the NRAMP1 gene can be regarded as a molecular marker for genetic selection of disease susceptibility in pig breeding.

• Asian-Australasian Journal of Animal Sciences
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• 제16권12호
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• pp.1830-1836
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• 2003

Studies using natural resistance-associated macrophage protein (NRAMP) identification indicated that cattle could be selected for immunity. Several studies performed on intracellular organisms such as Mycobacterium, Salmonella, Brucella and Leishmania in human and mouse revealed that resistance against these bacteria was dependent on high activity of NRAMP1 in macrophages. However, hardly any researches have been done on Staphylococcus aureus in bovine mastitis, which is an intracellular organism and the main cause of bovine mastitis. The objectives of this study were to establish reverse transcriptase polymerase chain reaction (RT-PCR) methods, through which NRAMP1 mRNA expression could be compared and analyzed between mastitis-resistant and -susceptible cows. NRAMP1 gene and its expression were investigated using 20 cows (Holstein Friesian) in Korea. Cows were evenly split into two groups, with and without histories of clinical mastitis. Equivalent numbers of cows were randomly selected from each group. Monocytes were isolated from the bovine peripheral blood of each selected cows and activated with lipopolysaccharide (LPS). mRNA was separated from the monocytes and cDNA of NRAMP1 was synthesized and amplified using RT-PCR with amplification of $\beta$-actin as a control. The difference in NRAMP1 expressions of mastitis-resistant (n=10) and -susceptible (n=10) Holstein cows was analyzed. Results demonstrate that resistant cows produced more NRAMP1 mRNA than the susceptible ones, and ratios of NRAMP1:$\beta$-actin expression were higher in resistant cows with or without LPS activation. Therefore, this study could be applied to select bovine mastitis resistant cows before infection based on the expression of NRAMP1.

• Asian-Australasian Journal of Animal Sciences
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• 제17권9호
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• pp.1183-1187
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• 2004

The natural resistance-associated macrophage protein 1 (NRAMP1) gene was identified as a candidate gene controlling the resistance and susceptibility to a number of intracellular parasites in pigs. The genetic variations in a 1.6 kb region spanning exon 1 and exon 3 of the porcine NRAMP1 gene were investigated by PCR-HinfI-RFLP in samples of 1347 individuals from 21 Chinese indigenous pig populations and 3 western pig breeds. Three alleles (A, B, C) and four genotypes (AA, BB, AB, BC) were detected. Significant differences in genotype and allele frequencies were observed between Chinese indigenous pig populations and exotic pig breeds, while in general the differences in genotype and allele frequencies among Chinese indigenous pig populations were not significant. The allele C was detected only in Duroc, Leping Spotted and Dongxiang Spotted pig, and the two Chinese pig populations showed similar genotype and allele frequencies. Four Chinese Tibetan pig populations displayed genetic differentiation at the NRAMP1 gene locus. In addition, intron 5 of the NRAMP1 gene was isolated and characterized by directly sequencing the PCR products encompassing intron 5. The alignment of intron 5 of the porcine, human, equine and ovine NRAMP1 gene showed a similarity of 45.38% between pig and human, 52.55% between pig and horse, 63.47% between pig and sheep, respectively.

• Tuberculosis and Respiratory Diseases
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• 제62권6호
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• pp.492-498
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• 2007

연구배경: 결핵균에 노출된 후 임상적 결핵으로 발병하는데 유전적 인자가 관여할 수 있으며, 치료 반응에도 숙주의 유전적 요인이 관여할 가능성이 있다. 이에 저자들은 NRAMP1 유전자 다형성을 감수성결핵과 다제내성결핵환자로 나누어서 비교하였다. 방 법: 100명의 약제감수성군, 102명의 다제내성군, 96명의 건강대조군을 대상으로 274C/T, 469+14G/C, 577-18G/A, 823C/T, A318V, 1465-85G/A, D543N, 1729+55del4 의 NRAMP1 유전자 다형성의 빈도를 중합효소 연쇄반응기법과 중합효소 연쇄반응-제한분절길이 다형성법을 이용하여 분석하였다. 결 과: NRAMP1 유전자의 D543N 이형접합체의 빈도는 건강대조군에 비해 약제감수성군에서 높았으나(OR=2.10, 95% CI=1.00 to 4.41, p=0.049) 다제내성군에서는 차이가 없었다. 823C/T의 이형접합체의 빈도는 건강대조군에 비해 약제감수성군(OR=2.79, 95% CI=1.11 to 7.04, p=0.029)과 다제내성군 (OR=3.30, 95% CI=1.33 to 8.18, p=0.010)에서 유의하게 높았으나 약제감수성군과 다제내성군 사이에는 유의한 차이가 없었다. 결 론: NRAMP1 유전자의 823C/T 이형접합체의 빈도는 약제감수성군과 다제내성군에서 유의하게 높아, 폐결핵의 발생과 연관되는 후보유전자일 가능성이 있으며 약제감수성결과에 따른 결핵이환에는 차이를 보이지 않았다.

• Asian-Australasian Journal of Animal Sciences
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• 제23권8호
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• pp.1089-1095
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• 2010

Four biological candidate genes, natural resistance associated macrophage protein 1 (SLC11A1 or NRAMP), prosaposin (PSAP), interferon Gamma (IFNG), and toll-like receptor 4 (TLR4), were examined to identify single nucleotide polymorphisms (SNP) and associations of the SNP with antibody response kinetics in hens. An $F_2$ population was produced by mating $G_0$ highly inbred (<99%) males of two MHC-congenic Fayoumi lines with highly inbred Leghorn hens. The $F_2$ hens (n = 158) were injected twice with SRBC and whole, fixed Brucella abortus (BA). Blood samples were obtained before each immunization, at 7 d after primary immunization, and at several time points after secondary immunization. Minimum titers (Ymin) and the time needed to reach them (Tmin), and maximum (Ymax) titers and the time needed to reach them (Tmax), were estimated from the seven post-secondary immunization titers using a nonlinear regression model. The $F_2$ hens were genotyped for the four candidate genes by using PCR-RFLP for one SNP per gene, which identified the parental allele. General linear models were used to test associations of SNP genotypes with antibody response parameters and BW measured at 4 ages. The IFNG SNP was highly significantly (p<0.0125) associated with primary response to SRBC, Tmin to BA, Ymin to BA, and 12-week BW. The current study demonstrated that the novel IFNG promoter SNP was associated with antibody kinetics for BA and SRBC in laying hens, and also with BW, suggesting that this cytokine may play a pivotal role in the relationship between immune function and growth.