Development of a Gene Cloning and Inactivation System for Halorespiring Desulfitobacterium dehalogenans

机译：卤吸脱硫脱硫杆菌基因克隆和灭活系统的开发

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Efficient host-vector systems have been developed for the versatile, strictly anaerobic, halo- and fumarate-respiring gram-positive bacterium Desulfitobacterium dehalogenans. An electroporation-based transformation procedure resulting in approximately 10³ to 10⁴ transformants per μg of the cloning vector pIL253 was developed and validated. The broad-host-range vector pG⁺host9 was shown to replicate at a permissive temperature of 30°C, whereas the replicon was not functional at 40°C. The D. dehalogenans frdCAB operon, predicted to encode a fumarate reductase, was cloned, characterized, and targeted for insertional inactivation by pG⁺host9 carrying a 0.6-kb internal frdA fragment. Single-crossover integration at the frdA locus occurred at a frequency of 3.3 × 10⁻⁴ per cell and resulted in partially impaired fumarate reductase activity. The gene cloning and inactivation systems described here provide a solid basis for the further elucidation of the halorespiratory network in D. dehalogenans and allow for its further exploitation as a dedicated degrader.

机译：已经开发了用于多功能，严格厌氧，可吸入卤代和富马酸酯的革兰氏阳性细菌脱硫脱硫杆菌的高效宿主载体系统。开发并验证了基于电穿孔的转化程序，该程序可产生每μg克隆载体pIL253约10 ^{3 至10 ^{4 转化子。宽宿主范围的载体pG ^{+ host9显示在30°C的允许温度下复制，而复制子在40°C时不起作用。克隆，表征，表征，表征，表征，表征并通过携带0.6kb内部frdA片段的pG ^{+ host9插入灭活的D. dehalogenans frdCAB操纵子。 frdA位点的单交换整合发生频率为每个细胞3.3×10 ^{-4 ，导致富马酸酯还原酶活性部分受损。本文所述的基因克隆和失活系统为进一步阐明脱卤杜氏梭菌中的呼吸道网络提供了坚实的基础，并使其进一步用作专门的降解剂。}}}}}

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期刊名称 Applied and Environmental Microbiology
作者
Hauke Smidt; John van der Oost; Willem M. de Vos;
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作者单位

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年(卷),期 2001(67),2
年度 2001
页码 591–597
总页数 7
原文格式 PDF
正文语种
中图分类应用微生物学;微生物学;
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专利

1. Development of a Gene Cloning and Inactivation System for Halorespiring Desulfitobacterium dehalogenans [J] . Hauke Smidt, John van der Oost, Willem M. de Vos Applied and Environmental Microbiology . 2001,第2期

机译：卤吸脱硫脱硫杆菌基因克隆和灭活系统的开发
2. Development of canine X-chromosome inactivation pattern analysis for the detection of cell clonality by incorporating the examination of the SLIT and NTRK-like family member 4 (SLITRK4) gene [J] . Tomita A., Mochizuki H., Tsuboi M., Research in Veterinary Science . 2019,第期

机译：犬X-染色体的灭活模式分析通过纳入狭缝和NTRK形的家庭成员4（SLITRK4）基因的检查来检测细胞克隆
3. Cloning of a functional Salmonella SPI-1 type III secretion system and development of a method to create mutations and epitope fusions in the cloned genes [J] . Wilson JW, Nickerson CA Journal of Biotechnology . 2006,第2期

机译：功能性沙门氏菌SPI-1 III型分泌系统的克隆以及在克隆的基因中产生突变和表位融合的方法的开发
4. Generic code Cloning method for detection of Clone code in software Development [C] . Syed Mohd Fazalul Haque, V Srikanth, E. Sreenivasa Reddy Proceedings of 2016 International Conference on Data Mining and Advanced Computing . 2016

机译：在软件开发中检测克隆代码的通用代码克隆方法
5. Positional cloning and characterization of ALF4, a gene required for root system development in Arabidopsis thaliana. [D] . DiDonato, Raymond Joseph, Jr. 2002

机译：拟南芥根系发育所需的基因ALF4的位置克隆和鉴定。
6. The effects of Capn1 gene inactivation on skeletal muscle growth development and atrophy and the compensatory role of other proteolytic systems [O] . C. M. Kemp, W. T. Oliver, T. L. Wheeler, -1

机译：Capn1基因失活对骨骼肌生长发育和萎缩的影响以及其他蛋白水解系统的补偿作用
7. Development of a Gene Cloning and Inactivation System for Halorespiring Desulfitobacterium dehalogenans [O] . Smidt, Hauke, van der Oost, John, de Vos, Willem M. 2001

机译：卤吸脱硫脱硫杆菌基因克隆和灭活系统的开发
8. Development of a Gene Cloning System in Methanogens [R] . Nagle, D. P., McCarthy, D. 1988

机译：methanogens基因克隆系统的开发

Development of a Gene Cloning and Inactivation System for Halorespiring Desulfitobacterium dehalogenans

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