YEAST ESCHERICHIA-COLI SHUTTLE VECTORS WITH MULTIPLE UNIQUE RESTRICTION SITES
YEAST ESCHERICHIA-COLI SHUTTLE VECTORS WITH MULTIPLE UNIQUE RESTRICTION SITES
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DOI：
10.1002/yea.320020304
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发表时间：
1986-09-01
期刊：
影响因子：
2.6
通讯作者：
TZAGOLOFF, A
中科院分区：
文献类型：
--
作者：
HILL, JE;MYERS, AM;TZAGOLOFF, A
两种酵母/E.构建了 colishuttle 载体。 YEp351和YEp352这两个载体具有以下特性：（1）它们可以在酿酒酵母和大肠杆菌中自主复制；（2）它们含有β-内酰胺酶基因并赋予大肠杆菌氨苄青霉素抗性。大肠杆菌； (3)它们含有pUC18的完整序列； (4) pUC18多克隆区的EcoRI、SacI、KpnI、SmaI、BamH1、XbaI、SbaI、SalI、PstI、SphI和HindIII这10个酶切位点在YEp352中都是唯一的；除了EcoRI和KpnI出现两次外，这些位点在YEp351中也是独特的； (5)在YEp351和YEp352的多克隆区中插入DNA的重组质粒可以通过适当E中β-半乳糖苷酶功能的丧失来识别。大肠杆菌宿主； (6) YEp351和YEp352分别含有酵母LEU2和URA3基因，允许选择在非选择性条件下生长的这些基因，表明高质粒拷贝数。上述特性使穿梭载体适用于构建酵母基因组文库和克隆由大量不同限制性位点定义的DNA片段。这两种载体已通过删除酵母中自主复制所需的序列进行了进一步修饰。因此，衍生质粒 YIp651 和 YIp352 可用于将特定序列整合到酵母染色体 DNA 中。
Two yeast/E. colishuttle vectors have been constructed. The two vectors, YEp351 and YEp352, have the following properties: (1) they can replicate autonomuosly inSaccharomyces cerevisiae and in E. coli;(2) they contain the β‐lactamase gene and confer ampicillin resistance toE. coli; (3) they contain the entire sequence of pUC18; (4) all ten restriction sites of the multiple cloning region of pUC18 includingEcoRI,SacI,KpnI,SmaI,BamH1,XbaI,SbaI,SalI,PstI,SphIandHindIIIare unique in YEp352; these sites are also unique in YEp351 except forEcoRIandKpnI, which occur twice; (5) recombinant plasmids with DNA inserts in the multiple cloning region of YEp351 and YEp352 can be recognised by loss of β‐galactosidase function in appropriateE. colihosts; (6) YEp351 and YEp352 contain the yeastLEU2andURA3genes, respectively, allowing for selection of these grown under non‐selective conditions indicative of high plasmid copy number. The above properties make the shuttle vectors suitable for constructions of yeast genomic libraries and for cloning of DNA fragments defined by a large number of different restriction sites.The two vectors have been further modified by deletion of the sequences necessary for antunomous replication in yeast. The derivative plasmids YIp651 and YIp352 can therefore be used ti integrate specific sequences into yeast chromosomal DNA.