| 用曲细精管微注射法建立绿色荧光蛋白转基因小鼠 |
| |
| 引用本文： | 沈新明,乔贵林,张玲,江培洲,黄华,姚开泰.用曲细精管微注射法建立绿色荧光蛋白转基因小鼠[J].南方医科大学学报,2002,22(3):250-253. |
| |
| 作者姓名： | 沈新明 乔贵林 张玲 江培洲 黄华 姚开泰 |
| |
| 作者单位： | 第一军医大学肿瘤研究所,广东广州,510515 |
| |
| 摘 要： | 目的研究曲细精管微注射法建立转基因小鼠的可行性。方法选取人巨细胞病毒启动子调控的增强型绿色荧光蛋白(pCMV-EGFP)表达载体,应用体视解剖镜和显微注射仪将不同剂量的被脂质体包裹的质粒DNA注射到不同年龄的KM小鼠曲细精管内。在注射后40d左右,雄鼠与雌鼠合笼交配;分娩后3周,剪取仔鼠尾,提取基因组DNA,应用PCR、Southernblotting技术进行整合检测。处死2只首建小鼠,荧光显微镜观察组织冰冻切片中EGFP的表达。结果共注射41只雄性小鼠,存活34只,其中32只具有交配、受精能力,获得仔鼠382只。仔鼠经检测,PCR阳性133只,Southernblotting阳性15只。小鼠的年龄、注射剂量与EGFP的整合率没有明显关系,荧光显微镜观察下没有观察到组织冰冻切片中EGFP明显表达。结论曲细精管微注射法是动物基因转移的一条简便可行的新途径。
|
| 关 键 词： | 曲细精管微注射 绿色荧光蛋白 转基因小鼠 |
| 文章编号： | 1000-2588(2002)03-0250-04 |
| 修稿时间： | 2002年1月18日 |
Construction of transgenic mice carrying enhanced green fluorescent protein gene by seminiferous tubule microinjection |
| |
| SHENXin-ming,QIAOGui-lin,ZHANGLing,JIANGPei-zhou,HUANGHua,YAOKai-tai Instituteof CancerResearch,FirstMilitaryMedicalUniversity,Guangzhou,China.Construction of transgenic mice carrying enhanced green fluorescent protein gene by seminiferous tubule microinjection[J].Journal of Southern Medical University,2002,22(3):250-253. |
| |
| Authors: | SHENXin-ming QIAOGui-lin ZHANGLing JIANGPei-zhou HUANGHua YAOKai-tai Instituteof CancerResearch FirstMilitaryMedicalUniversity Guangzhou China |
| |
| Affiliation: | SHENXin-ming,QIAOGui-lin,ZHANGLing,JIANGPei-zhou,HUANGHua,YAOKai-tai Instituteof CancerResearch,FirstMilitaryMedicalUniversity,Guangzhou510515,China |
| |
| Abstract: | Objective To studythefeasibilityof establishingtransgenicmicecarryingenhancedgreenflourescentprotein(EGFP)geneby meansof seminiferoustubulemicroinjection.Methods Thevectorexpressingenhancedgreenfluorescent proteinunderthecontrolof humancytomegalovirusimmediate-earlypromoter(pCMV-EGFP)wasselectedandmixedwith liposomein vitro.Microinjectionat differentdosesof theliposome-entrappedplasmidDNAintotheseminiferoustubulesof malemiceatdifferentageswasperformedto establishtransgenicmice,whichweremadeto matewithfemalemiceatleast40d afterthemicroinjection.GenomicDNAwasextractedfromtheoffspringof thefoundermiceforPCRandSouthernblotting analysis,andthefrozensectionsof differenttissuesfrom2of thefoundersmicewerepreparedforfluorescencemicroscopic observation.Results Amongthe41micereceivingthemicroinjection,32survivedandretainedtheirmatingabilityand fertility,and amongtheir382offspring133werepositivefor EGFPDNAas demonstratedby PCR,15of whichwere confirmedby Southernblottinganalysis.Theageof themiceor thedosesof microinjectiontheyreceivedwas notshownto impacttheintegrationof EGFPgene,andfluorescencemicroscopyfailedto detectsignificantEGFPexpressioninthetissues of thefoundermice(P>0.05)incomparisonwithnormalmice.Conclusion Seminiferoustubulemicroinjectionis simpleand practicableto implementgenetransferinmice. |
| |
| Keywords: | seminiferoustubulesmicroinjection enhancedgreenfluorescentprotein genetransferrats |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
