肠型点状气单胞菌和鱼害粘球菌融合子的构建  

Establishment of fusants of Aeromonas punctta f. intestinalis and Myxococcs piscicola

作　　者：钟蕾[1] 肖克宇[1] 戴良英[2] 

机构地区：[1]湖南农业大学动物科学技术学院,湖南长沙410128 [2]湖南农业大学生物安全科学技术学院,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2010年第1期65-68,76,共5页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家科技支撑计划项目(2007BAD87B11)

摘　　要：为探索鱼类病原菌融合疫苗制备的可行性,以肠型点状气单细胞菌58-20-9菌株和鱼害粘球菌G4菌株作为初发菌株进行原生质体融合,构建融合子.在酶解温度37℃下,筛选出最适酶解时间为40min,最适溶菌酶质量浓度为4mg/mL.以PEG为促融剂的条件下,58-20-9菌株与G4菌株的原生质体进行了融合,58-20-9菌株的原生质体形成率为54.7%,原生质体再生率为18.2%,G4菌株的原生质体形成率为50.8%,原生质体再生率为12.2%,初筛的原生质体融合率为4‰.点种500个初筛融合子菌落在双抗培养基上连续传代12次,得到1个稳定的融合子菌落,其稳定率为0.2%.In order to grope for feasibility and basic theory of preparing fusants bacterins of fish pathogeny, strains, Aerornonas punctta f.intestinalis 58-20-9 and Myxococcs piscicola G4 were employed as arch-strains to conduct protoplast fusion and establish fusants. When the temperature of enzymolysis was 37 ℃, the best enzymolysis time was singled out to be 40 minutes and the best lysozyme density to be 4 mg/mL. Under the condition of operating PEG as agent of accelerating fusion,Aeromonas punctta f.intestinalis 58-20-9 strains and Myxococcs piscicola G4 strains performed a protoplast fusion, and their protoplast formation rates were 54.7%, 50.8% respectively and their protoplast regenerative rates of 58-20-9 strains were 18.2% and 12.2% respectively. Arch-screening protoplast fusion rates were 49/00. After dibbling seeds of 500 fusants colony from arch-screening on duplex repelling selected culture medium and generating continuously 12 times, a steady fusant colony could be derived, whose steady rates were 0.2%.

关 键 词：肠型点状气单胞菌 鱼害粘球菌 原生质体融合 草鱼 

分 类 号：Q813.2[生物学—生物工程] S917.1[农业科学—水产科学]