Jin C, Liu H, Yang S, Zhang S
Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Chin J Biotechnol. 1995;11(3):185-91.
Thermostable DNA polymerase genes had been amplified from Thermus aquaticus YT-1 using the PCR technique. The amplified 2.5-kb DNA fragment was inserted into pUC18 and confirmed to be a thermostable DNA polymerase gene by restriction mapping and DNA sequencing. The insert fragment was then combined into an expression vector, pBV221. This recombinant plasmid overexpressed a 94-kDa of recombinant protein in E. coli. A 100-mL E. coli. culture could yield 1.5 x 10(5) units of Taq DNA polymerase, which could be applied in the PCR.
利用聚合酶链式反应（PCR）技术从嗜热水生栖热菌YT-1中扩增出了热稳定DNA聚合酶基因。将扩增得到的2.5kb DNA片段插入pUC18载体，并通过限制性酶切图谱分析和DNA测序确认其为热稳定DNA聚合酶基因。随后将插入片段连接到表达载体pBV221中。该重组质粒在大肠杆菌中过量表达了一种94kDa的重组蛋白。100mL大肠杆菌培养物可产生1.5×10⁵单位的Taq DNA聚合酶，可应用于PCR反应。
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