Bioinformatics analysis and tissue expression detection of Chinese alligator CXCL8 gene
In order to further understand the function of CXCL8 in Chinese alligator and its role in the pathogenesis of gout,CXCL8 gene was cloned from the liver tissue of Chinese alligator,bioinforma-tics analysis,homology analysis and evolutionary tree construction were performed.A prokaryotic ex-pression vector was constructed to express CXCL8 protein,and then mouse anti-Chinese alligator CXCL8 polyclonal antibody was prepared.The transcriptional levels of CXCL8 in Chinese alligator at diffe-rent periods were measured and analyzed.The results showed that the length of ORF sequence of CXCL8 gene was 291 bp,encoding 96 amino acids and molecular weight was 10.67 ku.The results of homology anal-ysis and phylogenetic tree construction showed that CXCL8 gene of Chinese alligator had 95%homology with Alligator mississippiensis,had a strong affinity and was located in the same branch of the phylo-genetic tree.The prepared polyclonal antibody had a titer of 1 ∶102 400 and good immunogenicity.CXCL8 expression was highest in kidney and small intestine,and was higher at the end of hibernation than that at the beginning of hibernation.The expression results were similar to those in the high incidence pe-riod of alligator gout.The results laid a foundation for further understanding of the regulatory mech-anism of CXCL8 gene in Chinese alligator on gout,and provided new insights into the pathogenesis of Chinese alligator gout.
Chinese alligatorCXCL8sequence analysisprokaryotic expressionpolyclonal antibody
国家科技期刊平台
NSTL
万方数据
