【作者】 林崇韫； 贺聪； 陈峰； 毕英佐； 赵亚华； 谢青梅；

【Author】 LIN Chong-yun~2,He Cong~1,,Chen Feng~1,Bi Ying-zuo~1,Zhao Ya-hua~(2*),Xie Qing-mei~(1*) (College of Animal Science,South China Agricultural University,Guangzhou,510642;College of Life Science,South China Agricultural University,Guangzhou,510642)

【机构】 华南农业大学生命科学学院； 华南农业大学动物科学学院；

【摘要】 为实现外源抗菌多肽基因在鸡输卵管中的表达,抑制病原菌通过母鸡垂直传播给子代,本文克隆了鸡卵清蛋白（Ovalbumin,ov）5端的调控序列1.1kb,将之取代载体pVAX1的CMV启动子,并将蜂毒肽基因（melittin,MLT）插入其下游,构建了输卵管定位表达抗菌多肽重组载体pOV_1.1-MLT,将该重组质粒转染CHO细胞和导入鸡体内,皆成功检测到目的蛋白的表达,并显示出抗菌活性,实现了输卵管定位表达抗菌多肽基因。更多还原

【Abstract】 This study was aiming at investigating the specifically expressed antimicrobial peptides in epithelial cells of oviduct magnum,which secreted albumen.Antimicrobial peptides incorporating with eggs could inhibit the invasion of pathogens in the formation of eggs,and finally would present in egg white.By using this method,we could expect higher chicken’s birth rate and reduce the chances of infection before the eggs hatch.The chicken ovalbumin 5’-flanking reguratory region was amplified from the genome of chicken oviduct tissue of laying hens.The 1.1kb chicken ovalbumin 5 -regulatory region were subcloned into the pVAX1 vector,replacing CMV promoter.Then,the melittin（MLT） gene was inserted in downstream of the 5’-regulatory region to construct recombinant chicken oviduct-specific expression vectors named pOV_1.1-MLT.The recombinant plasmid vector mixed with liposome was transfected into CHO cells and injected into laying hens via wing vein.RT-PCR and immunofluorescence assay indicated that recombinant proteins MLT were expressed and showed antibacterial activity.It is demonstrated that antimicrobial peptides gene under the direction of chicken ovalbumin gene promoter could be expressed in vitro and vivo.更多还原