Pyle L E, Finch L R
Russell Grimwade School of Biochemistry, University of Melbourne, Parkville, Victoria, Australia.
Nucleic Acids Res. 1988 Jul 11;16(13):6027-39. doi: 10.1093/nar/16.13.6027.
A physical map is presented for the 1200 kb genome of Mycoplasma mycoides subsp. mycoides Y, locating 32 cleavage sites for 8 restriction endonucleases. The large restriction fragments involved were separated and sized by pulsed-field agarose gel electrophoresis. Their locations on the map were determined by probing Southern blots of digests with individual fragments isolated from other digests and by correlating the products of double and triple digestions. Loci for 2 ribosomal RNA operons and 2 tRNA operons have been determined by probing with cloned genes and the broad regions of the replication origin and terminus have also been outlined by in vivo labelling studies.
本文展示了丝状支原体丝状亚种Y株1200 kb基因组的物理图谱，确定了8种限制性内切酶的32个切割位点。通过脉冲场琼脂糖凝胶电泳分离并测定了所涉及的大限制性片段的大小。通过用从其他酶切消化物中分离出的单个片段探测酶切消化物的Southern杂交印迹，并通过对比双酶切和三酶切产物，确定了这些片段在图谱上的位置。通过用克隆基因进行探测，确定了2个核糖体RNA操纵子和2个tRNA操纵子的基因座，并且通过体内标记研究也勾勒出了复制起点和终点的大致区域。
