期刊
ACS SYNTHETIC BIOLOGY
卷 7, 期 2, 页码 384-391出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.7b00377
关键词
cyanobacteria; biocontainment; CO2-concentrating mechanism; carboxysome; horizontal gene transfer; natural competence
资金
- National Science Foundation [EFRI-1240268]
- Department of Energy [DE-SC0010329]
- William F. Vilas Trust
- National Institutes of Health (NIH) Biotechnology Training Fellowship [NIGMS-5 T32 GM08349]
- Emerging Frontiers & Multidisciplinary Activities
- Directorate For Engineering [1240268] Funding Source: National Science Foundation
- U.S. Department of Energy (DOE) [DE-SC0010329] Funding Source: U.S. Department of Energy (DOE)
As researchers engineer cyanobacteria for biotechnological applications, we must consider potential environmental release of these organisms. Previous theoretical work has considered cyanobacterial containment through elimination of the CO2-concentrating mechanism (CCM) to impose a high-CO2 requirement (HCR), which could be provided in the cultivation environment but not in the surroundings. In this work, we experimentally implemented an HCR containment mechanism in Synechococcus sp. strain PCC7002 (PCC7002) through deletion of carboxysome shell proteins and showed that this mechanism contained cyanobacteria in a 5% CO2 environment. We considered escape through horizontal gene transfer (HGT) and reduced the risk of HGT escape by deleting competence genes. We showed that the HCR containment mechanism did not negatively impact the performance of a strain of PCC7002 engineered for L-lactate production. We showed through coculture experiments of HCR strains with ccm-containing strains that this HCR mechanism reduced the frequency of escape below the NIH recommended limit for recombinant organisms of one escape event in 10(8) CFU.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据