费氏中华根瘤菌 USDA193 的重复 DNA 序列的存在及共生质粒（pSym）的部分限制酶切图谱。

The presence of repeated DNA sequences and a partial restriction map of the pSym of Rhizobium fredii USDA193.

作者信息

Masterson R V, Atherly A G

出版信息

Plasmid. 1986 Jul;16(1):37-44. doi: 10.1016/0147-619x(86)90077-6.

Abstract

The large, 350-kb Sym (symbiotic) plasmid pRjaUSDA193 of Rhizobium fredii was examined to determine the frequency of repeated sequences present and to produce a physical and genetic map of a large region of the plasmid. A novel hybridization method, the Southern Cross, revealed that the plasmid pRjaUSDA193 contained many repeated sequences and assisted in restriction enzyme mapping of a 100-kb region containing nod genes. A cosmid clone bank was prepared with the broad-host-range cosmid pVK102. The restriction enzymes HindIII, HpaI, and KpnI were used to construct a physical map of overlapping clones. Labeled nod gene sequences were used to determine their location in the mapped region.

摘要

对费氏中华根瘤菌的大型共生质粒pRjaUSDA193（大小为350 kb）进行了检测，以确定重复序列的出现频率，并构建该质粒大片段区域的物理图谱和遗传图谱。一种新的杂交方法——Southern Cross，揭示了质粒pRjaUSDA193含有许多重复序列，并辅助对包含结瘤基因的100 kb区域进行了限制性酶切图谱分析。利用广宿主黏粒pVK102构建了黏粒克隆文库。使用限制性内切酶HindIII、HpaI和KpnI构建了重叠克隆的物理图谱。用标记的结瘤基因序列确定它们在图谱区域中的位置。