摘要
根据毕赤酵母(Pichia pastoris)偏爱密码子对设计的新型抗菌肽CAMa进行基因合成,将合成的CAMa基因克隆至载体pPICZαA上,构建穿梭载体pPICZαA-CAMa,电转化酵母菌X-33,经甲醇诱导表达,测定重组CAMa的抗菌活性,并通过断奶仔猪饲喂试验评价抗菌肽CAMa代替抗生素的效果。结果表明:新型抗菌肽CAMa在毕赤酵母X-33中获得分泌表达,并且对金黄色葡萄球菌CowanⅠ、大肠杆菌K12D31、猪大肠杆菌K88、猪大肠杆菌K99、猪大肠杆菌987P和猪沙门氏菌C782均有较好的抑杀活性。抗菌肽CAMa试验组和硫酸抗敌素对照组在仔猪增重、平均日增重、腹泻发生率上无显著差异(P>0.05),可作为抗生素类饲料添加剂替代品使用。
The gene of a novel designed antibacterial peptide CAMa was synthesized by using the preferential condon of P.pastoris and cloned into vector pPICZa-A to form pPICZaA-CAMa,which was transformed to P.pastoris X-33 by electroporation.Positive recombinants were screened for expression under induction of methanol.The expressed CAMa protein was determined for antibacterial activity.Antibacterial peptide CAMa was fed to the weaned piglets as an antibiotic alternative.Results showed that the antibacterial peptide CAMa was successfully expressed in P.pastoris X-33 which showing strong antibacterial activities against to 5.aureus Cowan I,E.coli K12 D31,E.coli K88,E.coli K99,E.coli 987 P and Salmonella C782 respectively.No significant difference in weight gain,average daily gain and diarrhea rate was observed between CAMa groups and colistin control groups(P>0.05).The above results showed that the CAMa can be used as additive of the feedstuff for piglets instead of antibiotic.
作者
凌红丽
郭佳宏
夏叶
蒋贻海
魏波
缪德年
LING Hong-li;GUO Jia-hong;XIA Ye;JIANG Yi-hai;WEI Bo;MIAO De-nian(Qingdao Vland Biotech Group Co.,Ltd.,Qingdao 266001,China;Shanghai Engineering Research Center of Breeding Pig,Shanghai Academy of Agricultural Sciences,Shanghai201106,China)
出处
《上海农业学报》
2019年第6期82-87,共6页
Acta Agriculturae Shanghai
基金
十二五国家科技支撑计划(2015BAD11B02-03)
上海工程技术研究中心建设专项(16dz2250700)
关键词
抗菌肽CAMa
毕赤酵母
抗菌活性
替代抗生素
Antibacterial peptide CAMa
Pichia pastoris
Antibacterial activity
Antibiotic alternative
