鸭疫里默杆菌与大肠杆菌融合株的构建

Establishment of the protoplast fusion cells of Riemerella anatipestifer and Escherichia coli

对鸭疫里默杆菌(YL1)和大肠杆菌(ZYD2)原生质体的制备、再生、融合条件进行优化,成功获得了64株具有双亲菌株耐药性,能够同时凝集兔抗ZYD2血清和兔抗YL1血清,且能够稳定遗传的融合菌株;通过培养特性、菌落特征、融合菌株形态染色特性、生化试验等鉴定,筛选出4株典型融合菌株DL4、DL5、DL8、DL29。攻毒试验表明,ZYD2的半数致死量为1.5×107,YL1的半数致死量为5.0×107,4株典型融合菌株的半数致死量分别为2.8×108、1.7×108、1.0×109、4.4×109,融合菌株毒性比亲本菌株毒性减弱。免疫保护试验和免疫鸭抗体水平测定结果证实,融合菌株具有良好免疫原性。

This experiment optimized the conditions of formation,regeneration and recombination of protoplas in Riemerella anatipestifer（YL1） and Escherichia coli（ZYD2）,successfully got 64 strains with parents＇ drug resistance and genetic stability,which could both agglutinated the rabbit anti-ZYD2 serum and the rabbit anti-YL1 serum.Thorough the identification of culture characteristics,colony characteristics,protoplast fusion morphology,dyeing properties,biochemistry experiment,we screened 4 typical protoplast strains DL4,DL5,DL8,DL29.The challenge test showed that ZYD2＇ LD50 was 1.5×107,YL1＇ LD50 was 5.0×107,4 typical protoplast strains＇ LD50 was 2.8×108,1.7×108,1.0×109,4.4×109 respectively.Toxicity of protoplast fusion is decreased.It was proved that the fusion strain had nicer immunogenicity with the immunity protect test and antibody level mensuration in the immuned ducks.