A CLONING VECTOR ABLE TO REPLICATE IN ESCHERICHIA-COLI AND STREPTOCOCCUS-SANGUIS
A CLONING VECTOR ABLE TO REPLICATE IN ESCHERICHIA-COLI AND STREPTOCOCCUS-SANGUIS
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DOI：
10.1016/0378-1119(82)90025-7
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发表时间：
1982-01-01
期刊：
影响因子：
3.5
通讯作者：
CLEWELL, DB
中科院分区：
文献类型：
--
作者：
MACRINA, FL;TOBIAN, JA;CLEWELL, DB
通过体外连接pACYC184 （Cmr - Tcr）和pVA749 （Emr）复制子构建了能在大肠杆菌和血链球菌中复制的质粒。该质粒被命名为pVA838，大小为9.2 kb[千碱基]，在大肠杆菌和血链球菌中表达Emr。其Cmr标记仅在大肠杆菌中表达，可通过在其内部EcoRI或PvuII位点添加DNA插入物而灭活。pVA838分子还含有适合分子克隆的独特的SalI、SphI、BamHI、NruI和XbaI切割位点。pVA838可以在大肠杆菌中扩增，但在血链球菌中不能扩增。以pVA838质粒为穿梭载体，克隆大肠杆菌链球菌质粒片段。这种从大肠杆菌中分离出来的嵌合体很容易通过转化引入血链球菌。
A plasmid that is able to replicate in E. coli and S. sanguis was constructed by the in vitro joining of the pACYC184 (Cmr Tcr) and pVA749 (Emr) replicons. This plasmid, designated pVA838, is 9.2 kb [kilobase] in size and expresses Emr in E. coli and S. sanguis. Its Cmr marker is expressed only in E. coli and may be inactivated by addition of DNA inserts at its internal EcoRI or PvuII sites. The pVA838 molecule also contains unique SalI, SphI, BamHI, NruI and XbaI cleavage sites suitable for molecular cloning. pVA838 may be amplified in E. coli but not in S. sanguis. The pVA838 plasmid was used as a shuttle vector to clone streptococcal plasmid fragments in E. coli. Such chimeras isolated from E. coli were readily introduced into S. sanguis by transformation.