High-Efficiency Expression Of VP2Gene Of Infectious Bursal Diease Virus In Yeast And The Optimization Of The Fermentation Process

Infectious bursal disease is one of the unsolved problems in the world. If chickensare infected by IBD, the bursa will be atrophied, even blooding. The chickens alsowill lose immunity. Once, the chicken get the IBD, the mounts of chickens will die.As the genes of Infectious bursal disease virus often change, the IBD is difficult tocontrol. So this disease is inquired by researchers In all countries.The thesis contains two parts. One part is studying that the genes of VP2of IBDVswitch to yeast and studying that the expression of VP2protein in yeast. The other isstudying the optimization of the fermentation process.(1) The genes of cDNA of VP2protein switch to the genes of VP2by thetechnology of RT-PCR.(2) The structure of PMD18-VP2: the genes of VP2connect to PMD18-T(3) First, I design two primers by premier5.0. Then, using the PMD18-VP2astemplate, I switch the genes of VP2to Ppicza. So the structure of Ppicza-VP2isestablished.(4) I switch the genes of VP2to yeast’s chromosomes.(5) To confirm that the genes of VP2have switched to cloning vector、expressionvector and yeast, I use the technology of the enzyme’s cut and send the genes to thecompany of the genes of analysis. The results of the genes of analysis is that thelength of the genes of VP2is1.5kb.(6) I use the single factor experiment to confirm the best level of the fourfactors(Temperature、pH、Rotate Speed and the adding amount of inducers) Theresults are25℃、4.0、190r/min、3%. (7) I use the orthogonal experiment to confirm that the key factor ofinfluencing the expression of VP2protein is the amount of inducers. I also get thebest level from the experiment-A2B3C2.(8) I use the ultraviolet spectrophotometry to get the amount of the expressionof VP2protein-o.6554g/L. The expression is high efficiency.