<i>GBP6</i>: differential expression in pulmonary alveolar macrophages under PRRSV infection and association with blood parameters of its missense mutation

Abstract

The GBP6 gene belongs to the gene family of interferon-gamma inducible GTPases, and is located in a previously characterized QTL region of porcine reproductive and respiratory syndrome virus (PRRSV) resistance in SSC4. In the current study, RT-PCR assay revealed that the GBP6 gene was widely distributed but highly expressed in spleen and lung. Under PRRSV infection, the GBP6 gene was up-regulated and differentially expressed in the pulmonary alveolar macrophages (PAMs) of Large White and Tongcheng pigs. Multiple sequences alignment and phylogenetic analysis revealed that GBP6 is highly conserved across species. We validated and analyzed the polymorphism of a missense mutation (n.932A>G, rs322187731) in the F2 resource population of Duroc × Erhualian, which changed the Valine (V) to Methionine (M) in the 300th residue of porcine GBP6. Through a linear mixed effects model, the polymorphism of the mutation (n.932A>G, rs322187731) was significantly associated with five haematological traits, namely platelet count at 20 days (PLT-20), plateletcrit at 20 days (PCT-20), white blood cell at 33 days (WBC-33), neutrophil count at 33 days (NE-33) and red blood cell count at 80 days (RBC-80). The findings presented here provide basic information for the function and possible involvement of the GBP6 gene in immune regulation and disease resistance. ______________________________________________________________________________________

Association of single nucleotide polymorphism at long non-coding RNA 8138.1 with duration of fertility in egg-laying hens

“…TCTTAGCAGCA). The PCR-RFLP condition for amplifying was pre-denaturing at 95 °C for 3 min, and 32 cycles of denaturation at 95 °C for 30 s, annealing at 64 °C for 30 s, extension at 72 °C for 20 s, and elongation at 72 °C for 7 min (Adeyinka et al, 2017). Next, PCR-RFLP products were digested with an enzyme ( Hind III ) and electrophoresed using 3% agarose gel.…”

&lt;i&gt;RAI14&lt;/i&gt; in the blood feather regulates chicken pigmentation

“…According to the NanoDrop measurements, all RNA samples with optical density (OD) 260/280 ratio of 1.9-2.0 and OD 260/230 ratio of >1.8 were reverse transcribed to cDNA using PrimeScript™ RT reagent kit (Takara Biotechnology Co. Ltd.) according to manufacturer recommendations. The 25 µL qPCR reaction contained 12.5 µL 2× SYBR qPCR mix (Aidlab Biotechnologies Co., Ltd., China), 0.5 µL (10 µM) of each primer, and 1 µL cDNA (Adeyinka et al, 2017). Polymerase chain reaction (PCR) amplification was carried out using a Bio-Rad Laboratories, Inc. device at 95 • C for 2 min, followed by 40 cycles at 95 • C for 10 s and 60 • C for 15 s. The qPCR amplifications were conducted using an independent set of 10 biological and 3 technical replicates per sample.…”