杏花鸡白细胞介素10(IL--10)基因的克隆及其表达分析

Cloning and Expression Analysis of Interleukin-10(IL-10) Gene in Xinghua Chicken

为研究杏花鸡白细胞介素10(IL-10)基因在抵抗鸡球虫病中的作用,应用PCR、基因克隆和测序等技术获得杏花鸡完整的IL-10基因CDS序列,进行杏花鸡IL-10基因CDS序列和核苷酸序列与其他物种间的同源性分析,比较在QM-7成肌细胞、鸡肠上皮细胞IEC、鸡巨噬细胞HD11和鸡成纤维细胞DF-1中IL-10基因的表达差异以及在不同肠道组织中IL-10的表达量差异。结果表明,完整地扩增了IL-10基因的完整CDS序列,发现了3个同义突变(162A>G、180T>C和303C>T)。构建的发育树表明杏花鸡IL-10基因CDS序列和核苷酸序列与红色原鸡、鹌鹑、火鸡和珍珠鸡的同源性最高。IL-10在鸡肠上皮细胞的表达量最高,在盲肠扁桃体中表达量最高。球虫子孢子的入侵导致DF-1细胞和HD11细胞中的IL-10表达量升高。研究结果为IL-10基因在肠道免疫和抵抗球虫病的研究提供理论基础。

In order to study the role of interleukin-10(IL-10)gene in resisting chicken coccidiosis, the complete CDS sequence of IL-10 gene in Xinghua chicken was obtained by PCR, gene cloning and sequencing. Homology analysis of the CDS sequence and nucleotide sequence of IL-10 gene in Xinghua chicken and other species were performed. The expression differences of IL-10 in different cells and intestinal tissues were verified. The results showed that the complete CDS sequence of IL-10 gene was completely cloned, and three synonymous mutations(162 A>G, 180 T>C and303 C>T) were found. The phylogenetic tree indicated that the CDS sequence and nucleotide sequence of the IL-10 gene of Xinghua chicken had the highest homology with Gallus gallus, Coturnix japonica, Meleagris gallopavo and Numida meleagris. IL-10 has the highest expression in chicken intestinal epithelial cells and cecal tonsil. The infection of coccidia spores resulted in increased expression of IL-10 in DF-1 cell and HD11 cell.The results provide a theoretical basis for further study on IL-10 gene in intestinal immunity and resistance to coccidiosis in chicken.