Knowledge Resource Center for Ecological Environment in Arid Area
DOI | 10.1186/s12917-023-03709-x |
Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes | |
Souza, Jorge Lucas Nascimento; Silva, Fernanda Cavalcante; da Silva, Carlikelly Gleicy; Bomfim, Isabela Maria Fortaleza Neves; de Medeiros, Henrique Rocha; Zaros, Lilian Giotto | |
通讯作者 | Zaros, LG |
来源期刊 | BMC VETERINARY RESEARCH
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EISSN | 1746-6148 |
出版年 | 2023 |
卷号 | 19期号:1 |
英文摘要 | Analyzing the stability of reference genes already described as universal is an important methodology to lead gene expression analysis because different studies have shown that the expression of universal reference genes may vary between experimental treatments. In this sense, the glyceraldehyde 3-phosphate dehydrogenase (GAPDH), Succinate dehydrogenase complex subunit A (SDHA) and Ribosomal Protein L-19 (RPL-19) reference genes (already described in other studies with sheep from different regions, breeds and infectious agents or in organisms evolutionarily close to sheep) were investigated in the abomasum, small and large intestines of resistant and susceptible crossbred sheep groups to gastrointestinal nematode infections in the Semi-arid region in Northeast of Brazil. The animals were naturally infected to determine the resistance or susceptibility status by counting eggs per gram (EPG) of feces from the gastrointestinal tract after 33 weeks of observations of infection evolution. Relative gene expression was performed by RT-qPCR methodology using Sybr green and relative gene expression stability was tested by different software programs such as REST, BestKeeper, geNorm and Normfinder. Our results showed the susceptible animals had increase in egg counts per gram of feces than resistant animals (p < 0.001), and both groups showed a mixed infection by nematodes of the genus Haemonchus, Trichostrongylus, Oesophagostomum and Trichuris. Furthermore, we show the importance of analyzing different genes in different software programs and the importance to choose ideal reference genes. In this sense, GAPDH was the most stable gene in the abomasum, whereas SDHA was the most stable in the small and large intestines. In addition, we discuss about variables which can interfere in relative expression such as breed, species, climate and tissue. However, utilizing other reference genes already described in other studies with the same and different variables should be performed. |
英文关键词 | GAPDH Gastrointestinal tract Sheep and RT-qPCR |
类型 | Article |
语种 | 英语 |
开放获取类型 | gold |
收录类别 | SCI-E |
WOS记录号 | WOS:001064981600003 |
WOS关键词 | TIME QUANTITATIVE PCR ; HAEMONCHUS-CONTORTUS ; EXPRESSION ANALYSIS ; RT-PCR ; RESISTANCE ; MECHANISMS ; IVERMECTIN ; SELECTION ; COOPERIA ; HISTORY |
WOS类目 | Veterinary Sciences |
WOS研究方向 | Veterinary Sciences |
资源类型 | 期刊论文 |
条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/395597 |
推荐引用方式 GB/T 7714 | Souza, Jorge Lucas Nascimento,Silva, Fernanda Cavalcante,da Silva, Carlikelly Gleicy,et al. Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes[J],2023,19(1). |
APA | Souza, Jorge Lucas Nascimento,Silva, Fernanda Cavalcante,da Silva, Carlikelly Gleicy,Bomfim, Isabela Maria Fortaleza Neves,de Medeiros, Henrique Rocha,&Zaros, Lilian Giotto.(2023).Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes.BMC VETERINARY RESEARCH,19(1). |
MLA | Souza, Jorge Lucas Nascimento,et al."Analysis of the stability of the reference genes GAPDH, SDHA and RPL-19 in sheep from a semi-arid region infected by gastrointestinal nematodes".BMC VETERINARY RESEARCH 19.1(2023). |
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