Cloning, Sequence Analysis And Detection By In Situ Hybridization Of Chicken Interleukin-1 β CDNA

According to the described sequence of chicken interleukin-1β(ChIL-1β) and by using the extracted total RNA of chicken peripheral blood mononuclear cell (PMBC), a pair of primers were designed to reverse transcriptionally amplify and to compare the expressional degrees of ChIL-1β mRNA by stimulating chicken PMBC for 4,8,12,24 hour with lipopolysaccharide (LPS) and for 24 hour with LPS and actidione respectively. The results showed that stimulating chicken PMBC with LPS and actidione to produce ChIL-1β was a better way of artifical stimulation.Connecting the production of RT-PCR with the plasmid vector of pUC19 and transforming the vector into Ecoli bacillus, the recombinational bodies of ChIL-1β were produced. Then they were examined by PCR amplifying, enzyme digesting and sequence analysis. Analysis of ChIL-1βshowed that it had more than 99% homology identity to the described ChIL- 1β sequence.The double chains of ChIL-1β cDNA were labeled with Dig-11-dUTP to produce probe . And the ChIL-1β mRNA secreation was studied in the tissues of chickens' brains infected with AEV and that of normal chickens'. The results showed that the content of ChIL-1βsecreation in sick chickens' brain was higher than that of the normal chickens'.