鸡β-防御素-3在毕赤酵母中的分泌表达

Expression and secretion of chicken β-defensin-3 in Pichia Pastoris

对鸡β-防御素-3(Gal-3)在毕赤酵母中的分泌表达进行了研究。以重组质粒Gal-3-T为模板,利用PCR技术扩增出Gal-3基因成熟肽片段,将该片段插入到酵母表达载体pPICZα-C,构建分泌型重组表达载体pPICZα-C-Gal-3,电转入表达宿主菌X-33毕赤酵母,Zeocin抗性筛选重组菌株;挑取阳性菌株经甲醇诱导表达,Tricine-SDS-PAGE电泳发现在约4.5 kDa位置出现预期条带;对所表达的Gal-3进行抗菌活性检测,发现其具有抗大肠杆菌、沙门氏菌、金黄色葡萄球菌等细菌的活性。

The mature fragment of chicken β-defensin-3（Gallinacin-3） gene was amplified from the plasmid of Gal-3-T by polymerase chain reaction （PCR）. The PCR products were inserted into the pPICZα-C expression vector. The recombinat vector named pPICZα-C-Gal-3 was electro-transformed into pichia pastoris X-33 strain. The positive transformed strains were cultured and induced by addition of methanol. Expression products with molecular weight approximately 4.5 kDa were displayed on the gel during Tricine-SDS-PAGE. The expression products had antimicrobial activity against E. coli, salmonella pallorum and ataphylococcosis aureas.