Characterization of the Basic Replicon ofRhodococcus Plasmid pSOX and Development of aRhodococcus-Escherichia coli Shuttle Vector
Characterization of the Basic Replicon ofRhodococcus Plasmid pSOX and Development of aRhodococcus-Escherichia coli Shuttle Vector
复制标题
红球菌质粒pSOX基本复制子的表征及红球菌-大肠杆菌穿梭载体的开发
DOI：
10.1128/aem.64.11.4363-4367.1998
复制
发表时间：
1998
影响因子：
4.4
通讯作者：
P. Lau
中科院分区：
文献类型：
--
作者：
Claude Denis;H. Bergeron;D. Labbé;C. Greer;J. Hawari;M. Grossman;B. M. Sankey;P. Lau
摘要对红球菌100kb脱硫质粒(PSOX)的复制区进行了分析。将X309菌株定位于一个4kbKpnI片段，并测定了其序列。其中一个预测的开放阅读框架(ORF)的氨基酸序列与分枝杆菌pLR7质粒家族的假定复制(Rep)蛋白序列相关。在预测的5个ORF产物中，有3个是通过大肠杆菌T7聚合酶/启动子系统的放射性标记进行鉴定的。在大肠杆菌中，pSOX的Rep蛋白显然是以21.3 kDa的形式合成的，而不是预测的41.3 kDa，这是内部引发的结果。这种情况让人想起一些细菌代表蛋白的情况。构建了含有pSOX基因、pBluescrip II KS−和pRF29氯霉素抗性(Cmr)基因的穿梭载体。用这种新的穿梭载体证明了枯草芽孢杆菌SACB基因在一株红球菌中的表达，使其对蔗糖的存在很敏感。
ABSTRACT The replication region of a 100-kb desulfurization plasmid (pSOX) from Rhodococcus sp. strain X309 was localized to a 4-kbKpnI fragment, and its sequence was determined. The amino acid sequence of one of the predicted open reading frames (ORFs) was related to the putative replication (Rep) protein sequences of the mycobacterial pLR7 family of plasmids. Three of the five predicted ORF products were identified by radiolabelling with the Escherichia coli T7 polymerase/promoter system. In E. coli, the Rep protein of pSOX was apparently synthesized in a shortened form, 21.3 kDa instead of the predicted 41.3 kDa, as a result of an internal initiation. This situation is reminescent of that for some bacterial Rep proteins. A shuttle plasmid was constructed with the pSOX origin, pBluescript II KS−, and the chloramphenicol resistance (Cmr) gene from pRF29. This new shuttle plasmid was used to demonstrate expression of the Bacillus subtilis sacB gene in a strain of Rhodococcus, rendering it sensitive to the presence of sucrose.
含有 iteron 的 DNA 分子复制的调节。
DOI：
10.1016/s0079-6603(08)60857-0
发表时间：
1994
期刊：
Progress in nucleic acid research and molecular biology
影响因子：
--
作者：
Filutowicz,M;Dellis,S;Levchenko,I;Urh,M;Wu,F;York,D
通讯作者：
York,D
影响因子：
56.9
作者：
RAVICHANDRAN, KG;BODDUPALLI, SS;DEISENHOFER, J
通讯作者：
DEISENHOFER, J
影响因子：
5.7
作者：
HASEMANN, CA;KURUMBAIL, RG;DEISENHOFER, J
通讯作者：
DEISENHOFER, J