| 摘要: |
| 为了获取具有广泛适用性的DNA聚合酶，将深海热液区获得的超嗜热古菌Palaeococcus pacificus DY20341的PolB DNA聚合酶和dUTPase进行原核表达和纯化，将得到的PolB DNA聚合酶及dUTPase混合后用于扩增古菌、细菌和真核生物的基因.通过PCR反应成功地扩增了商业化Pfu DNA聚合酶所不能扩增的Palaeococcus pacificus DY2034的琼脂酶基因.由此可知，尽管商业化的DNA聚合酶多种多样，针对不同扩增目的片段所需的特异DNA聚合酶体系仍然是必要的，这是又一嗜热古菌B家族DNA聚合酶应用的报道. |
| 关键词: 海洋生物学 Palaeococcus pacificus PolB DNA聚合酶 dUTPase 原核表达 PCR |
| DOI：10.3969/J.ISSN.2095-4972.2016.02.007 |
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| 基金项目:厦门市海洋与渔业局资助项目（14GZP65NF29）；国家海洋局第三海洋研究所基本科研业务费专项资金资助项目（海三科2014021） |
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| PCR application of thermostable DNA polymerase B and dUTPase from hyprothermophilic archaea Palaeococcus pacificus |
| CHEN Zimeng,ZHAO Wenzhao,XIE Bingbin,CUI Zhizhong,LI Zhuo |
| (Animal Science and Technology College，Shandong Agricultural University，Taian 271018，China;Key Laboratory of Marine Biogenetic Resources，Third Institute of Oceanography，SOA，Xiamen 361005，China;School of Life Sciences，Xianmen University，Xiamen 361005，China) |
| Abstract: |
| A new family B DNA polymerase （ppa_PolB） from thermophilic archaea strain Palaeococcus pacificus DY20341 was recombinantly expressed and purified. The dUTPase （ppa_dUTPase） from the strain was also obtained. Using a serial of ppa_PolB and ppa_dUTPase combinations， we tested their performance in amplifications of fragments from archaea， bacteria and eukaryote. One of the combinations successfully amplified the Palaeococcus pacificus DY2034 six agarase genes， which could not be achieved by commercial DNA polymerase. We concluded the necessary of expanding DNA polymerases for variety of goals in PCR due to this new DNA polymerase combination． |
| Key words: marine biology Palaeococcus pacificus PolB DNA polymerase dUTPase prokaryotic expression PCR |
