耐热DNA聚合酶基因的克隆及在大肠杆菌中的表达

用PCR法从水生栖热菌菌株YT－1中扩增耐热DNA聚合酶基因，得到2．5kb的DNA片段t扩增片段重组到pUCl8中测序证实为Taq DNA聚合酶基因，将该片段重组到pBV221温控表达质粒中，在大肠杆菌中表达出94kDa的重组蛋白，100ml培养物的细胞产酶为1．5×10⁵u，表达的蛋白能催化PCR反应的进行。

Cloning and Expression of Thermostable DNA Polymerase Gene in Escherichia coli

Thermostable DNA polymerase gene had been amplified from Thermus aquaticus YT-1 using PCR technique. The amplified 2. 5kb DNA fragement was inserted into pUCl8 and confirmed to be thermostable DNA polymerase gene by restriction mapping and DNA sequencing. The insert fragement was then combined into an expression vector, pBV221. This recombinant plasmid overexpressed a 94kDa of recombinant protein in E. colt. 100ml of E. colt culture could yield 1. 5X 105 units of Taq DNA polymerase, which could be applied in the PCR.