| 地芽孢杆菌属DYth03 DNA聚合酶基因的克隆、表达及性质分析 |
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| 引用本文： | 罗淑娅,李侃,徐丽美.地芽孢杆菌属DYth03 DNA聚合酶基因的克隆、表达及性质分析[J].台湾海峡,2011,30(1):17-20. |
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| 作者姓名： | 罗淑娅 李侃 徐丽美 |
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| 作者单位： | 国家海洋局第三海洋研究所、国家海洋局海洋生物遗传资源重点实验室,福建,厦门,361005 |
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| 基金项目： | 国家863计划资助项目 |
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| 摘 要： | 从东太平洋热液区E53站位的深海沉积物样品中分离出1株能在65℃生长的嗜热菌（DYth03）.该菌的16S rDNA序列与地芽孢杆菌属（Geobacillus）内各种之间的同源性为98%以上.克隆得到DYth03的DNA聚合酶基因（DYth-pol）,序列分析表明该基因全长为2 631 bp,G＋C含量为55.5%,推测编码为876个氨基酸,与BstDNApolI的同源性最高（达98%）.将该聚合酶基因克隆到pTTQ-h表达载体上,并在大肠杆菌DH1中进行表达.对纯化到的表达产物进行酶活性测定,结果表明该酶具有聚合酶活性和5’-3’外切酶活性.
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| 关 键 词： | 海洋生物学 嗜热菌 地芽孢杆菌属（Geobacillus） 16S rDNA 耐热DNA聚合酶 基因表达 东太平洋 |
Cloning, expression and characterization of the DNA polymerase gene from Geobacillus sp.DYth03 |
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| LUO Shu-ya,LI Kan,XU Li-mei.Cloning, expression and characterization of the DNA polymerase gene from Geobacillus sp.DYth03[J].Journal of Oceanography In Taiwan Strait,2011,30(1):17-20. |
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| Authors: | LUO Shu-ya LI Kan XU Li-mei |
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| Institution: | (Key Laboratory of Marine Biogenetic Resources/Third Institute of Oceanography,SOA,Xiamen 361005,China) |
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| Abstract: | A strain of thermophilic bacteria(DYth03) that can grow at 65℃ was isolated from the sediment of a hydrothermal vent field on the E53 site of the east Pacific Ocean.The 16S rDNA of DYth03 shared more than 98% sequence homology with related Geobacillus species.The gene encoding the DNA polymerase of DYth03(DYth-pol) was cloned and sequenced.Analysis indicated that the gene was 2 631 bp in length with a G+C content of 55.5%,encoding an 876-amino-acid peptide,which was most similar to that of Bst DNApolI(98%).The DYth-pol gene was cloned in the expression vetor pTTQ-h and expressed in E.coli DH1.The expressed product was purified and characterized.It showed 5’-3’ exonuclease and polymerase activity. |
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| Keywords: | 16S rDNA |
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