摘要
目的 将抗菌肽D基因克隆至甲醇营养型酵母表达载体pPICZα A中 ,并与α factor信号肽相连接 ,经EcoRⅠ酶切分析和PCR扩增鉴定 ,获得阳性重组子。方法 用锂盐法将重组表达载体转化巴斯德毕赤酵母 (Pichiapastoris)受体菌GS115 ,筛选获得阳性克隆。结果 将阳性菌落发酵产物进行抑菌试验 ,表达产物有较强的杀菌作用 ,表达产物杀菌活性达到5 6 5 6U ml。结论 抗菌肽D基因已经导入酵母细胞并整合在其基因组中 ,表达产物均具有高效的杀菌作用 ,有希望开发成为一种新型的抗菌消毒药物。
Objective Cecropin D gene was cloned into methanol nutritional vector-pPICZα-A, linked with α-factor signal peptide. Methods Positive recombinant was gotten through PCR and EcoR Ⅰ restriction enzyme. The positive recombinant vector was transformed into Pichia pastoris yeast strain GS115.Results The positive recombinant yeast velum was fermented and its expressive product was assayed by the method of anti-germ. Conclusion Cecropin D gene was cloned into yeast cell and its expressive products has high activity of killing bacteria, Which could be developed into a new type of anti-bacterial medicine.
出处
《卫生研究》
CAS
CSCD
北大核心
2004年第1期81-85,共5页
Journal of Hygiene Research
基金
广东省科委高新技术孵化项目资助 (No .98ff14 )
关键词
抗菌肽
基因
毕赤酵母
表达
cecropin, gene, Pichia pastoris yeast, expression
