【作者】 龙淼； 李鹏； 张燚； 刘国文； 朱连勤； 王哲；

【Author】 LONG Miao1,2 Li Peng 1,2 Zhang Yi 1,2 LIU Guo-wen2~* ZHU Lian-qin 3 Wang Zhe2 (1 Shenyang Agricultural University,Shenyang 110866;2 College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062 3 Qingdao Agricultural University,Qingdao 266109)

【机构】 沈阳农业大学； 吉林大学畜牧兽医学院； 青岛农业大学；

【摘要】 目的 :对瘤胃分离鉴定的反刍兽月形单胞菌原生质体制备与再生的最佳条件进行确定,为反刍兽月形单胞菌的基因操作、基因组改组和原生质体融合做准备.方法 :从影响原生质体形成与再生的菌龄、酶浓度、酶解时间、酶解温度的四因素三水平上进行L9(34)正交试验.结果 :菌体培养12h,溶菌酶浓度为600μg/ml,酶解温度为30℃,酶解时间为30min时是反刍兽月形单胞菌原生质体制备与再生的最佳条件,并且原生质体的制备率和再生率分别为94%和17.2%.结论 :反刍兽月形单胞菌原生质体制备和再生条件的确立为对其进行基因操作、基因组改组和原生质体融合奠定基础.更多还原

【Abstract】 Objection:The optimum composition condition for the production and regeneration of Selenomonas ruminantium protoplasts are studied for gene manipulation,genome shuffling and protoplast fusion.Method:The factors on protoplast formation and regeneration such as cell age,enzyme concentration,enzymolysis time and temperature were studied through L9 (34) orthogonal experiment.Result:When cell age was 12h,using 600μg/ml of lysozyme,enzymolysis at 30℃for 30ain,the protoplasts rate of forming and regenerating was 94%and 17.2%.Conclusion:The establishment of the optimum composition condition for the production,genome shuffling and regeneration of Selenomonas ruminantium protoplasts provided the groundwork for the gene manipulation and protoplast fusion in the future.更多还原