Construction Of High Efficient Expression Strain Of Bovine Interferon

Interferon is a kind of highly active and multifunctional glycoprotein which has antiviral,antitumor and immune regulation effects induced by specific cells under the action of interferon inducer.Interferon is highly species-specific and can inhibit multiple RNA viruses as well as multiple DNA viruses.Bovine interferon is divided into two types,type I interferon and type II interferon,according to its type of action.Type I interferon is mainly divided into five subtypes of ?,?,?,? and ?.Type II interferon mainly has Gamma interferon.BoIFN-? mainly has antiviral function,and its antiviral ability is the strongest in interferon,which has a good therapeutic and preventive effect on animal viral diseases.Therefore,the study of bovine interferon is of great significance for the prevention and treatment of animal viral diseases.Four expression vectors were successfully constructed in this experiment:(1)This experiment successfully cloned the bovine interferon gene and ligated it to the pET-28 a vector.After sequencing,the result was completely correct.(2)In order to increase the expression of the protein,the interferon sequence was optimized by using the codon preference of E.coli without changing the amino acid sequence of the interferon,and was ligated to the pET-28 a vector.(3)In order to increase the protein expression level,the elements were expressed in tandem on pET-28a-BoIFN-?-t,and the sequenced expression elements were successfully ligated to pET-28a-BoIFN-?.(4)In order to improve the soluble expression of protein,the BoIFN-? gene was ligated into the low-temperature expression vector expression vector pET-28a-sumo,and the results were completely correct.BoIFN-? was successfully expressed by prokaryotic expression system,and high-expression strains have been screened to improve protein expression by optimizing sequences,tandem expression elements,improving medium components,and changing induction conditions.In order to increase the soluble expression of the protein,the low temperature expression vector pET-28a-sumo was selected to compare the soluble expression of the protein.It was determined that the optimal induction condition of BoIFN-? was optimized.The sequence of IPTG was 1mmol/L at 37°C in LB medium,and induced at 180 rpm for 4 hours,and the expression amount was 34.5% of total protein.The use of a low temperature expression vector can increase the soluble expression of the protein,but the total protein expression is also reduced.