Genomic organization and polymorphisms detected by denaturing high-performance liquid chromatography of porcine SLC11A1 gene.
SLC11A1 (also known as Natural Resistance Associated Macrophage Protein1, NRAMP1) plays a crucial role in resistance of inbred mice to infection with several intracellular pathogens such as Mycobacterium, Leishmania and Salmonella. In this study, PCR amplification and sequencing were performed to obtain the genomic organization and sequence of porcine SLC11A1 gene by comparative genomic analysis. Results showed that porcine SLC11A1 gene consists of 15 exons and 14 introns, which is consistent with that of mice and human. All introns were sequenced and their nucleotide sequences were submitted to GenBank. The exon/intron boundaries were determined by comparing cDNA sequence with amplified genomic DNA sequences. Mutational analysis was performed on exonic and neighboring intronic region by denaturing high-performance liquid chromatography (DHPLC) and sequencing confirmation. Forty polymorphisms were identified; six are located in exons and thirty-four in introns. Two exonic polymorphisms are nonsynonymous changes (D6H and V175I), three are synonymous changes (S23, G33 and I155), and one is in 3' UTR. The availability of the fine genomic organization and identification of the polymorphisms will facilitate the evaluation of porcine SLC11A1 functional role in diseases resistance or susceptibility.
Wu
Zhen-Fang
ZF
College of Animal Science, South China Agricultural University, Guangzhou, People's Republic of China. wzf@scau.edu.cn
Luo
Wen-Hua
WH
Yang
Guan-Fu
GF
Zhang
Xi-Quan
XQ
D000818:Animals; D001483:Base Sequence; D027682:Cation Transport Proteins / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004252:DNA Mutational Analysis; D017931:DNA Primers; D018076:DNA, Complementary; D005091:Exons; D016678:Genome; D007438:Introns; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D011110:Polymorphism, Genetic; D017422:Sequence Analysis, DNA; D034421:Sus scrofa; D000099173:Natural Resistance-associated Macrophage Protein 1
D027682:Cation Transport Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D000099173:Natural Resistance-associated Macrophage Protein 1