含蚕豆叶绿体rRNA基因重组质粒物理图谱的构建

PHYSICAL MAPPING OF RECOMBINANT PLASMIDS CONTAINING BROAD BEAN CHLOROPLAST RIBOSOMAL RNA GENES

以 pBR322 DNA 为载体,Escherichia coli HB101为受体菌,克隆了含蚕豆叶绿体 rRNA基因的二个 BamHI 片段。应用几种限制性内切酶酶切以及 Southern 印迹法构建了这二个特异片段的物理图谱。重组质粒 pVFB16含有一个4.70kb 的 BamHI 片段,其上含有完整的16S rRNA 基因;重组质粒 pVFB32含有一个5.65kb 的 BamHI 片段,其上含有23S rRNA基因,23S—4.5S/5S rRNA 基因的间隔区及4.5S/5S rRNA 基因。

TWO BamHI fragments containing broad bean chloroplast rRNA genes were cloned using the bacterial plasmid pBR322 as a vector and Escherichia coli HB101 as host bacterial.Physical maps of the two cloned ct DNA BamHI fragments containing rRNA genes were constructed by cleavage with several restriction endonucleases and Southern blot hybridization with E.coli 168—23S rRNAs.Recombinant plasmids pVFB16 and pVFB32 contain a 16SrRNA sequence on the 4.70 kb BamHI fragment,a 23S rRNA sequence and 4.5S/5S rRNA sequences on the 5.65 kb BamHI fragment,respectively.