双酶消化法组建昆虫病毒DNA物理图谱规则的探讨

双酶消化法组建昆虫病毒DNA物理图谱规则的探讨 THREE-THREE RULES FOR MAPPING INSECT VIRUS DNA BY DOUBLE ENDONUCLEASE DIGESTIONS 齐义鹏 Qi Yipeng Huang Youngxui 1 first-author 黄永秀 1 武汉大学病毒及分子生物学系 武汉大学病毒及分子生物学系 Department of Virology and Moleular Biology,Wuhan University Department of Virology and Moleular Biology,Wuhan University 在用双酶消化法组建 DNA 物理图谱的过程中,我们总结了一套"三三规则"即:三类片段（消失片段 Fd,保留片段 Fr,接头片段 Fl）;三种片段关系（Fd 1 =Fd 2 ,Fdi=1/2 Fl,Fl 1 =Fl 2 ）,三种排列（Fl 的首尾排列,Fr 的夹心排列,Fd 的相嵌排列）.用这一套规则组建了大尺蠖核型多角体病毒4个限制内切酶的物理图谱。其片段顺序分别是:BamHI,HCIDEBGFA;Xhol,AJBCEIHFDG;HindⅢ,AFBCMHLJKGIED;BglⅡ,BEAG-DFC. The insect baculovius BsNPV DNA was mapped with double endonuclease digestions.Using BamHI-H frament containing BsNPV polyhedrin gene serve as zero site,the order of the four restrition fragments are determined as follows:BglⅡ,BEAGDFC;BamHI,HCIDEBGFA;HindⅢ,AFBCMHLJ- KGIED;XhoⅠ,AJBCEIHFDG.Here we present a three three rule for restriction mapping of DNA based on our experiences and practices.They are:three type fragments,e.g,disappearance fragment （Fd）,retained frag- ment（Fr） and liker fragment（Fl）;three type aligments;e.g,head tail for Fl,sandwich for Fr,and mosaic for Fd;three relations among fragments, e.g,Fd 1 =Fd 2 ,Fd i =1/2ΣFl i ;Fl 1 =Fl 2 .This rule seems to have simplified the method and procedure for the construction of DNA restriction map. 大尺蠖核型多角体病毒 DNA 物理图谱 双酶消化 Inseet baculovirus DNA restriction map Double digestions 国家科学基金;国家教委资助的课题 1990-02-01 2021-04-01 2