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Development of field application vectors for bioremediation of soils contaminated with polychlorinated biphenyls.

机译:开发用于多氯联苯污染土壤的生物修复的现场应用载体。

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摘要

Field application vectors (FAVs), which are a combination of a selective substrate, a host, and a cloning vector, have been developed for the purpose of expressing foreign genes in nonsterile, competitive environments in which the gene products provide no advantage to the host. Such gene products are exemplified by the enzymes for the cometabolism of polychlorinated biphenyls (PCBs) through the biphenyl degradation pathway. Attempts to use highly competent PCB-cometabolizing strains in the environment in the absence of biphenyl have not been successful, while the addition of biphenyl is limited by its human toxicity and low water solubility. Broad-substrate-specificity PCB-degradative genes (bphABC) were cloned from a naturally occurring isolate. Pseudomonas sp. strain ENV307, into broad-host-range plasmid pRK293. The resulting PCB-degrading plasmids were transferred to the FAV host Pseudomonas paucimobilis 1IGP4, which utilizes the nontoxic, water-soluble, nonionic surfactant Igepal CO-720 as a selective growth substrate. Plasmid stability in the recombinant strains was determined in the absence of antibiotic selection. PCB-degrading activity was determined by resting cell assays. Treatment of contaminated soil (10, 100, or 1,000 ppm of Aroclor 1242) by surfactant amendment (1.0% [wt/wt]Igepal CO-720 in wet soil) and inoculation with recombinant isolates of strain 1IGP4 (approximately 4 x 10(6) cells per g of soil) resulted in degradation of many of the individual PCB congeners in the absence of biphenyl.(ABSTRACT TRUNCATED AT 250 WORDS)
机译:为了在非无菌,竞争性环境中表达外源基因而表达外源基因,已开发了由选择性底物,宿主和克隆载体组合而成的现场应用载体(FAV) 。此类基因产物的实例是通过联苯降解途径进行多氯联苯(PCB)的新陈代谢的酶。在没有联苯的情况下尝试在环境中使用功能强大的多氯联苯代谢菌株并未成功,而联苯的添加受到其人体毒性和低水溶性的限制。从天然分离物中克隆了广泛的底物特异性PCB降解基因(bphABC)。假单胞菌株ENV307,进入宽宿主范围的质粒pRK293。将所得的PCB降解质粒转移至FAV宿主pseudomonas paucimobilis 1IGP4,该宿主利用无毒的水溶性非离子表面活性剂Igepal CO-720作为选择性生长底物。在没有选择抗生素的情况下确定重组菌株中的质粒稳定性。通过静止细胞测定法确定PCB降解活性。通过表面活性剂改良剂(湿土壤中1.0%[wt / wt] Igepal CO-720)处理污染的土壤(Aroclor 1242为10、100或1,000 ppm),并用菌株1IGP4的重组分离株接种(约4 x 10(6) )/ g土壤中的细胞数)导致在没有联苯的情况下许多单个PCB同系物的降解。(摘要截断为250字)

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