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当前位置: 首页 > 文章 > 抗菌肽Thanatin转基因小鼠建立的初步研究 中国兽医学报 2010,30 (3) 299-303
Position: Home > Articles > Preliminary research on production of transgenic mice carrying thanatin gene Chinese Journal of Veterinary Science 2010,30 (3) 299-303

抗菌肽Thanatin转基因小鼠建立的初步研究

作  者：
张鑫;尹逊河;苗向阳;曲朝杰;张秋婷;马艳芳
单  位：
山东农业大学;中国农业科学院
关键词：
抗菌肽;睾丸注射;转基因鼠
摘  要：
人工合成抗菌肽Thanatin基因的3条片段,利用重叠延伸PCR扩增技术得到完整的Thanatin基因,通过分子克隆方法构建出pIRES2-EGFP-Thanatin重组表达载体。然后与脂质体混合,采用睾丸打点注射将新构建的抗菌肽基因注入小鼠睾丸内,共注射公鼠5只。6周后与雌鼠交配,对新生仔鼠进行断尾,提取尾尖基因组DNA,利用PCR和Southern印迹方法对其进行检测。结果显示,得到的52只新生仔鼠中PCR检测阳性率为38.46%,Southern印迹检测阳性率为30.77%;在活体荧光成像系统下转基因小鼠呈现绿色荧光表达。通过睾丸注射法使Thanatin基因在小鼠的基因组中得到整合,为进一步研究抗菌肽的作用机理、培育抗病动物品种以及通过建立转基因动物生物反应器进行抗菌肽的大量生产奠定了基础。
译  名：
Preliminary research on production of transgenic mice carrying thanatin gene
作  者：
ZHANG Xin1,2,YIN Xun-he1,MIAO Xiang-yang2,QU Zhao-jie1,2,ZHANG Qiu-ting2,MA Yan-fang1 (1.Animal Science and Technology,Shandong Agricultural University,Tai'an,Shandong 271018,China;2.Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100094,China)
关键词：
antibacterial peptides;intratesticular injection;transgenic mice
摘  要：
Thanatin gene was constructed by overlap extension PCR and inserted into pIRES2-EGFP vector.Then recombinant plasmid pIRES2-EGFP-Thanatin and liposome were mixed and injected into male mouse testis.Six weeks after injection,male mice mated with female mice.Genomic DNA was extracted from pups tail.Polymerase chain reaction(PCR) and Southern-blot were applied to identify transgenic mice.Fifty two pups were born and the positive ratio was respectively 38.46% and 30.77% by PCR and Southern-blot analysis.The result showed that thanatin gene was integrated into mice genomic DNA.Expression of green fluorescence protein in transgenic mice was observed in fluorescence imager.In the present study,establishment of transgenic mice carrying thanatin gene is attractively valuable and prospective in application.And this provides a basis for disease-resistant animal breeding and production of antibacterial peptides through transgenic animals.

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