Methods and Compositions for Limiting Viability of a Modified Host Cell Outside of Designated Process Conditions
a technology of host cell and composition, which is applied in the field of molecular biology, can solve the problems of insufficient evaluation of the consequences of releasing transgenic microorganisms into the environment, limited information about the ecology and evolution of transgenic microorganisms in the wild, etc., and achieve the effect of reducing the expression of gene and reducing the viability of the cell
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example 1
Genetically Modified Organisms Sensitive to Ultraviolet (UV) Radiation
[0056]Under controlled production conditions of a photosynthetic organism, UV radiation is filtered out of the incident radiation, but in the wild, UV exposure is unavoidable in the normal course of exposure to unfiltered sunlight. Mutations in the genes provided in Table 1 confer UV sensitivity. Prior to entering an industrial process, the organism is engineered to have reduced expression of a gene from Table 1. The engineered organism is robustly viable only under the controlled conditions of the process. In one embodiment, engineered organism maintains a gene from Table 1 until a designated treatment is given, i.e., exposure to the presence or absence of a certain chemical inducer, exposure to a defined temperature profile, etc.
[0057]Genes specific for UV resistance, such as those encoding photolyase, are selected for deletion. Genes are excised using a specific and complete excision method such as the λ att sy...
example 2
Genetically Modified Organisms Requiring High Levels of CO2
[0058]Under controlled production conditions of a photosynthetic organism, higher levels of CO2 are present than in the wild. Mutations in the genes provided in Table 2 decrease the ability of a cell to fix carbon from CO2 and render the cell unable to grow outside of the process condition. Prior to entering an industrial process, the organism is engineered to have reduced expression of a gene from Table 2. The engineered organism is robustly viable only under the controlled conditions of the process. In one embodiment, engineered organism maintains a gene from Table 2 until a designated treatment is given, i.e., exposure to the presence or absence of a certain chemical inducer, exposure to a defined temperature profile, etc.
TABLE 2Gene targets in Synechococcus sp. PCC 7002 for mutationor excision to reduce viability in natural CO2 conditions.LocusGene(s)Enzyme ActivitySYNPCC7002_A2547ndhBNAD(P)H-quinoneoxidoreductaseSYNPCC...
example 3
Methods for Genetically Modifying Organisms to Insert a Gene of Interest
[0061]Controllable site-specific integration into and excision from the host chromosome using a bacteriophage. Many bacteriophages, in particular λ-like bacteriophage, are capable of site-specific integration into and excision from the host chromosome. Integration generally requires the activity of a single protein, Int, which recognizes 2 pairs of recombination sequences (attL and attR), one pair of which resides on the phage chromosome and the other on the host chromosome. Recombination between these pairs of sequences creates a pair of hybrid sequences consisting of phage and chromosome sequences. Excision of the phage from the host generally requires at least one additional phage protein in addition, Xis, as well as the host encoded protein IHF, that allow the cell to recombine at the hybrid phage / chromosome sequence. The Int protein from bacteriophage HK022, which is similar to λ, has been shown to promote ...
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