分子克隆过程中基因组DNA的电镜观察技术的研究  

 Electron Microscopic Observation of Genomic DNA in Clone Procedure

作　　者：徐均焕[1] 冯明光[1] 

机构地区：[1]浙江农业大学生物科学系

出　　处：《电子显微学报》1998年第3期280-284,共5页Journal of Chinese Electron Microscopy Society

基　　金：国家杰出青年科学基金

摘　　要：为了尝试用电子显微镜对病毒外壳蛋白基因克隆过程中基因组ＤＮＡ的检测，我们采用电镜大分子展层技术制备电镜样品并观察了病毒外壳蛋白基因组ＤＮＡ经ＰＣＲ扩增，克隆到ＰＢＳ质粒ＤＮＡ中的全过程，电镜观察表明：ＰＢＳ质粒基因组ＤＮＡ有超螺旋状、环状、或断裂成线状，长度约为１．００μｍ，外壳蛋白基因为线状，长度为０．２０μｍ，外壳蛋白基因克隆到ＰＢＳ质粒后，重组子质粒ＤＮＡ有成环状或线状，长度为１．２０μｍ。电镜观察结果与其它生化实验结果相符。In order to observe genomic DNA in clone procedure with electron microscope,the EM molecular display technique has been used.Using TMV RNA as template,TMV cDNA was synthesized by AMV reverse transcriptase.The synthesized cDNA was then amplified by polymerase chain reaction (PCR).It was cloned in PBS Plasmid. Electron microscopic observation proved that three kinds of PBS DNA:superhelix DNA,open circular DNA or linear DNA were found,they were about 1 00μm in length,the virus coat protein genome in rod shaped was about 0.20μm in length,when the virus coat protein genomic DNA was cloned in PBS,the recombinant DNA in superhelix,circular or linear was about 1.20μm in length.Our experiment conformed to the other biochemical results.

关 键 词：电镜 大分子展层技术 DNA 分子克隆过程 

分 类 号：Q785[生物学—分子生物学]