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邻苯二酚-2，3-双加氧酶基因在芘降解菌基因组的整合与表达

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赵亚光, 段魏魏, 徐苗, 吴盼云, 肖璐梅, 马腾飞, 晁群芳. 邻苯二酚-2，3-双加氧酶基因在芘降解菌基因组的整合与表达[J]. 环境工程学报, 2019, 13(1): 232-237. doi: 10.12030/j.cjee.201807117
引用本文: 赵亚光, 段魏魏, 徐苗, 吴盼云, 肖璐梅, 马腾飞, 晁群芳. 邻苯二酚-2，3-双加氧酶基因在芘降解菌基因组的整合与表达[J]. 环境工程学报, 2019, 13(1): 232-237. doi: 10.12030/j.cjee.201807117
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ZHAO Yaguang, DUAN Weiwei, XU Miao, WU Panyun, XIAO Lumei, MA Tengfei, CHAO Qunfang. Integration and expression of catechol-2,3-dioxygenase gene in pyrene degrading bacteria genome[J]. Chinese Journal of Environmental Engineering, 2019, 13(1): 232-237. doi: 10.12030/j.cjee.201807117
Citation: ZHAO Yaguang, DUAN Weiwei, XU Miao, WU Panyun, XIAO Lumei, MA Tengfei, CHAO Qunfang. Integration and expression of catechol-2,3-dioxygenase gene in pyrene degrading bacteria genome[J]. Chinese Journal of Environmental Engineering, 2019, 13(1): 232-237. doi: 10.12030/j.cjee.201807117
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邻苯二酚-2，3-双加氧酶基因在芘降解菌基因组的整合与表达

  • 1.

    新疆大学生命科学与技术学院，乌鲁木齐 830046

  • 基金项目:

    国家自然科学基金资助项目(31460027)

Integration and expression of catechol-2,3-dioxygenase gene in pyrene degrading bacteria genome

  • 1.

    College of Life Science and Technology, Xinjiang University, Urumqi 830046, China

  • Fund Project:

  • 摘要: 为了构建能够稳定遗传且高效降解多环芳烃的工程菌，利用PCR技术对Pseudomonas songnenensis wp3-1的邻苯二酚-2, 3-双加氧酶(C23O)基因进行克隆，并将其与自杀性载体pUTmini-Tn5连接，得到重组载体pUTmini-Tn5-C23O。在三亲接合作用下，经mini-Tn5转座子将重组载体pUTmini-Tn5-C23O中的C23O基因整合到菌株Pseudomonas sp. wp4的染色体DNA中，最终得到基因工程菌wp4-C23O。在不同pH、温度下，菌株wp4和工程菌wp4-C23O对浓度为50 mg?L-1的芘进行降解7 d。2株菌降解最适温度为37 ℃、最适pH为7.5。在此条件下，工程菌wp4-C23O对芘降解率显著高于wp4菌株(P<0.05)，降解率提高11.45%。以PAHs降解优势菌株为受体构建工程菌可以去除石油污染土壤中的PAHs。
    Abstract: This study aims to construct polycyclic aromatic hydrocarbons engineering bacteria with genetic stability and highly efficient degradation. Firstly, the catechol-2,3-dioxygenase (C23O) gene of Pseudomonas songnenensis wp3-1 was cloned by PCR technique, and was linked to suicide vector pUTmini-Tn5, then the recombinant vector pUTmini-Tn5-C23O was obtained. Secondly, the C230 gene of pUTmini-Tn5-C23O was integrated into the chromosomal DNA of Pseudomonas sp. wp4 with the triparental conjugation effect of mini-Tn5 transposition, and finally the gene engineering bacteria wp4-C23O was obtained. The result of contrast experiment to degrade pyrene solution (50 mg?L-1) for 7 days indicated that the optimum environment conditions for strain wp4 and strain wp4-C23O were 37 ℃ and pH 7.5. Under these conditions, the engineering strain wp4-C23O showed significantly higher degradation rate of pyrene than wp4 strain (P<0.05), and it increased by 11.45%. The engineering bacteria taking PAHs degrading dominant strain as acceptor can effectively remove PAHs from petroleum-contaminated soil.
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  • 刊出日期:  2019-01-08

邻苯二酚-2，3-双加氧酶基因在芘降解菌基因组的整合与表达

  • 1. 新疆大学生命科学与技术学院，乌鲁木齐 830046
基金项目:

国家自然科学基金资助项目(31460027)

摘要: 为了构建能够稳定遗传且高效降解多环芳烃的工程菌，利用PCR技术对Pseudomonas songnenensis wp3-1的邻苯二酚-2, 3-双加氧酶(C23O)基因进行克隆，并将其与自杀性载体pUTmini-Tn5连接，得到重组载体pUTmini-Tn5-C23O。在三亲接合作用下，经mini-Tn5转座子将重组载体pUTmini-Tn5-C23O中的C23O基因整合到菌株Pseudomonas sp. wp4的染色体DNA中，最终得到基因工程菌wp4-C23O。在不同pH、温度下，菌株wp4和工程菌wp4-C23O对浓度为50 mg?L-1的芘进行降解7 d。2株菌降解最适温度为37 ℃、最适pH为7.5。在此条件下，工程菌wp4-C23O对芘降解率显著高于wp4菌株(P<0.05)，降解率提高11.45%。以PAHs降解优势菌株为受体构建工程菌可以去除石油污染土壤中的PAHs。

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