Analysis of SLC11A1 gene expression in healthy water buffalo (Bubalus bubalis) blood cells using qPCR

Abstract

ABSTRACT. SLC11A1 (solute carrier family 11 member 1 protein) gene influences the initial phase of bacterial cellular infections through macrophage activation. Recent literature on buffalo has attempted to associate the genotype of the polymorphic microsatellite located in the 3ꞌuntranslated region (3ꞌUTR) of the gene, with either susceptibility to brucellosis or with improved macrophage function. Carriers of the (GT) 16 allele have been reported to be resistant to brucellosis. In this study we analyzed the steady-state level of SLC11A1 expression in a serologically negative herd of 26 animals differing by the number of (GT) n microsatellite repeats by using a reverse transcriptase quantitative real-time polymerase chain reaction approach. We evaluated five different reference genes, which had not been reported previously, for use in gene expression experiments in buffalo blood. However, we did not find any significant difference between buffalo carriers of the different microsatellite alleles, with respect to SLC11A1 expression in whole blood or in blood fractions [peripheral blood mononuclear cells (PBMC) and polymorphonuclear leukocytes/granulocytes (PMN/G)]. Conversely, there was a difference between the blood fractions in their SLC11A1 expression levels, with the PMN/G fraction having a higher expression level than the PBMC fraction (P < 0.015).

Molecular comparison of Slc11a1 and Slc11a2 genes of swamp‐ and riverine‐type water buffaloes

“…However, the present study determined five amino acid substitutions in Slc11a1 of swamp‐type buffalo. Recent literatures on cattle (Martinez et al ., ; Paixao et al ., ) and buffalo (Borriello et al ., ; Crisa et al ., ) have attempted to associate with the polymorphic microsatellite located in the 3′untranslated region (3′UTR) of the gene. The 3′UTR region in Slc11a1 of cattle has no significant association with resistance to brucellosis in vivo (Paixao et al ., ), and this is in contrary in vitro (Martinez et al ., ).…”

Identification of the complete coding cDNAs and expression analysis of B4GALT1, LALBA, ST3GAL5, ST6GAL1 in the colostrum and milk of the Garganica and Maltese goat breeds to reveal possible implications for oligosaccharide biosynthesis

“…Primer Express software (Applied Biosystem, Thermo Fisher Scientific, Waltham, USA) was used to design primers for qPCR on the basis of bovine sequences. Five reference genes (RGs) selected in another study [68] were tested here: ATP synthase β polypeptide, a nuclear gene encoding a mitochondrial protein ( ATP5B ), eukaryotic translation initiation factor eIF-2B subunit β ( EIF2B2 ), succinate dehydrogenase complex subunit A flavoprotein ( SDHA ), DNA-directed RNA polymerase fragment RNA polymerase II ( POLR2A ), and a TATA box-binding protein ( TBP ). We designed and tested new primers for the following RGs: a ubiquitously expressed transcript ( UXT ) and ribosomal protein S9 ( RPS9 ).…”

Assessment of Poly(ADP-ribose) Polymerase1 (PARP1) expression and activity in cells purified from blood and milk of dairy cattle

“…The ATP synthase, H + transporting, mitochondrial F1 complex, beta polypeptide (ATP5B) and the mitochondrial ribosomal protein S15 (MRPS15) were tested for reference gene Table 1 Characteristics of the primer pairs used and the efficiency of amplification. suitability based on the findings of previous studies (Bionaz and Loor, 2007;Crisà et al, 2013). Information on PCR primer sets is summarized in Table 1.…”

“…1 (C, D) shows the relative expression of PARP1 mRNA in lymphocytes and monocytes compared to unsorted PBMC and normalized to MRPS15 and ATP5B. These two candidate reference genes (Table 1) were selected through the analysis of literature survey on studies in bovine with RT-qPCR analysis (Bionaz and Loor, 2007;Crisà et al, 2013 and some still unpublished results). They can be considered good as reference genes for the comparison of PARP1 expression in different cell types since their expression is stable with low coefficient of variation CV in different bovine samples tested in different experiments (data not shown) and there are no substantial differences in gene expression between PBMC, lymphocytes and monocytes, both for MRPS15 and ATP5B.…”