34 results about "Tetrahymena thermophila" patented technology

The ciliated protozoan Tetrahymena exemplifies a recombinant system for the expression of heterologous nucleic acids, preferably on the plasma membrane surface. Integration of a heterlogous nucleic acid into the β-tubulin gene, BTU1, of a paclitaxel-sensitive T. thermophila mutant that possesses btu1-IK350M β-tubulin allele allows screening for transformants using negative selection, as transformants have restored paclitaxel resistance. Transgenic ciliated protozoa of the invention can serve as live vaccines. For example, transgenic Tetrahymena expressing Ichthyophthirius multifiliis i-antigen protein on their surface are effective vehicles for vaccination of freshwater fish against infection by I. multifiliis.

A method characterizes samples having units by monitoring fluctuating intensities of radiation emitted, scattered, and / or reflected by the units in at least one measurement volume, the monitoring being performed by at least one detection device, the method comprising the steps of: a) measuring in a repetitive mode a number of photon counts per time interval of defined length, b) determining a function of the number of photon counts per the time interval, and c) determining a function of specific brightness of the units on basis of the function of the number of photon counts.

The invention discloses CYP5013C2 protein and coding gene thereof, and provides tetrahymena expressing the CYP5013C2 protein and an application of the tetrahymena in DDT degradation. According to the present invention, an application of the tetrahymena overexpressing the CYP5013C2 gene in DDT degradation is protected; the CYP5013C2 gene is the gene coding protein (a) or protein (B), wherein the protein (a) is protein comprising an amino acid sequence represented by a sequence 1 in the sequence table, and the protein (b) is the protein formed by carrying out substitution and / or deletion and / or addition of one or a plurality of amino acid residues on the amino acid sequence represented by the sequence 1, has the same function, and is derived from the sequence 1; and the recombinant tetrahymena has the following advantages that: (1) stress response of the CYP5013C2 gene to the DDT is specific and sensitive; (2) the CYP5013C2 genes involves in a DDT biotransformation process in tetrahymena cells; (3) the tetrahymena has an efficient DDT degradation function; (4) the DDT in a water body can be controlled in the bottom end of a food chain; and (5) environment pollution is not generated.

The invention discloses a tetrahymena transgenic carrier containing HSP70 promoter and GFP and preparation method and use thereof. Plasmid T-GFP is constructed by using plasmid rDNA-GFP in the methodssuch as enzyme cutting and PCR enzyme cutting site mutation; thermophile tetrahymena genome DNA is used for constructing plasmid T-HSP702 in the methods such as PCR augmentation and clone; Plasmids T-GFP, T-HSP702 and pD5H8 are used for construction and finally obtaining plasmid HSP702-GFP-pD5H8 by enzyme cutting and connection. The carrier contains HSP702 promoter, thereby being capable of realizing high-efficient expression of exogenous gene; the carrier contains replaceable exogenous gene gfp so that an exogenous gene expression system is realized for tetrahymena by replacing different exogenous genes; with the sieving effect of paromomycin medicine, the carrier replaces most rDNAs originally in the tetrahymena so that the exogenous genes are largely increased in the tetrahymena so asto realize genetic transformation of target genes. The method in the invention is easy and has convenient operation, and the carrier can be used for detection of environmental tributyl tin in transfection in tetrahymena.

The invention relates to an organic arsenic feed additive discharge pre-alarming monitoring method in a water environment system, belonging to the environmental monitoring technical field, comprising the following steps: preparing tetrahymena culture medium; preparing arsanilic acid standard sample solution / roxarsone standard sample solution; collecting the actual water sample and separating and extracting the organic arsenic feed additive compound; inoculating the tetrahymena into the prepared culture medium and then subpackaged the inoculated culture medium into the sample culture tubes; respectively adding the organic arsenic feed additive standard solution into the sample culture tubes and respectively measuring the corresponding tetrahymena cell density using absorptometry; making the actual water sample intervening the corresponding tetrahymena biological model and analyzing and determining the corresponding tetrahymena cell density; based on the comparison of the tetrahymena cell density and the contrast of the corresponding tetrahymena cell density after the actual water sample intervention, to obtain the concentration range of the pollutant presence, thereby pre-alarming the discharge condition of the organic arsenic feed additive in the corresponding actual water environment.

The invention discloses a construction method and a use of a polygene-transfected tetrahymena thermophila cell strain. The construction method comprises the following steps of cloning a homologous sequence of a target gene A in a targeting vector pNeo4, transforming tetrahymena in a nutritional stage by a particle bombardment method, carrying out paromomycin resistance screening to obtain a large nuclear gene-genetically transformed positive cell strain a, cloning a homologous sequence of a target gene B in a targeting vector pBsr, transforming the nutritional-stage tetrahymena cell strain a by the particle bombardment method, carrying out paromomycin-blasticidin combined resistance screening to obtain a large nuclear gene-genetically transformed positive cell strain a / b. The construction method solves the problem that the existing polygene transfection technology has low efficiency, a complicated screening process and long time, overcomes the limitation that the existing technology does not produce an essential gene-less or -mutant cell strain, and can be used for researching the essential gene-loss or -mutation-caused influence on related gene expression and function. The construction method can be used for directional modification such as knockout, mutation or epitope tagging of two different target genes thereby realizing tetrahymena polygene combined application research.