CONSTRUCTION AND EXPRESSION OF HYBRID PLASMIDS CONTAINING THE ESCHERICHIA-COLI-GLY A GENE
CONSTRUCTION AND EXPRESSION OF HYBRID PLASMIDS CONTAINING THE ESCHERICHIA-COLI-GLY A GENE
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DOI：
10.1016/0378-1119(81)90148-7
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发表时间：
1981-01-01
期刊：
影响因子：
3.5
通讯作者：
STAUFFER, LT
中科院分区：
文献类型：
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作者：
STAUFFER, GV;PLAMANN, MD;STAUFFER, LT
将编码丝氨酸转羟甲基酶(SthM EC 2.1.2.1)的基因克隆到pACYC184载体中。重组质粒(Pgs1)含有一个13kb的[kb碱基对]EcoRI插入片段。遗传和生化实验表明，控制SthM合成的区域存在于插入片段上。携带携带GlyA基因的多拷贝质粒载体的菌株将该酶的产量提高了17-26倍。通过分析来自Pgs1的一组携带可转座卡那霉素抗性元件Tn5的随机插入的质粒，鉴定了插入片段上的GlyA基因。将原始插入片段克隆到pBR322中，证实了2.5kb的Sali-BclI片段携带GlyA基因。给出了该碎片的物理地图。
The E. coli glyA gene, encoding serine transhydroxymethylase (STHM EC 2.1.2.1) was cloned in the plasmid vector pACYC184. The recombinant plasmid (pGS1) contains a 13 kb [kilobase pairs] EcoRI insert. Genetic and biochemical experiments indicate that the region controlling STHM synthesis is present on the insert. Strains bearing multicopy plasmid vectors carrying the glyA gene overproduce the enzyme from 17- to 26-fold. The glyA gene was identified on the insert by analyzing a set of plasmids derived from pGS1 that carry random insertions of the transposable kanamycin resistance element Tn5. Cloning of segments of the original insert into plasmid pBR322 established that a 2.5 kb SalI-BclI fragment carries the glyA gene. A physical map of this fragment is presented.