An in vitro process for the production of closed linear deoxyribonucleic acid (DNA) comprises (a) contacting a DNA template comprising at least one protelomerase target sequence with at least one DNA polymerase in the presence of one or more primers under conditions promoting amplification of the template; and (b) contacting amplified DNA produced in (a) with at least one protelomerase under conditions promoting production of closed linear DNA. A kit provides components necessary in the process.

The present disclosure discloses a protein-based multifunctional molecular switch for antibody detection, and belongs to the field of protein detection. The molecular switch is a fusion protein including the following parts ligated sequentially from an N-terminus to a C-terminus: a part (1): SmBiT; a part (2): an epitope polypeptide capable of being specifically bound by the antibody to be detected; and a part (3): LgBiT. In the present disclosure, the molecular switch can be specifically recognized by an antibody through the epitope polypeptide, thereby affecting binding of the SmBiT and the LgBiT, and greatly changing a luciferase activity before and after to reflect a concentration level of the antibody. The molecular switch can be used to detect a 2019 novel coronavirus (SARS-CoV-2) with an accuracy and specificity close to 100%, which has an extremely desirable use value.

The present disclosure provides compositions and methods for detecting the presence of neutralizing antibodies in a sample. Unlike conventional assays, the methods provided herein do not require the use of live virus or virus pseudoparticles to identify neutralizing antibodies.

A biosensor for detecting nitrotoluenes. Two P. putida host populations (H-I and H-II) are engineered. H-1 undergoes fluorescence when a nitrotoluene is detected but it is also engineered to metabolize nitrotoluenes to toluene as its sole nitrogen-source. H-I is 1-ACC Deaminase inactive and is further engineered to efflux toluene and provide toluene to adjacent H-II. In H-II, ACC is the N-source and metabolizes toluene as the sole carbon and energy source available. The H-II cells are engineered to not be able to use medium fructose. The H-II population has a promoter/GFP construct with a promoter sensitive to toluene and thus they fluoresce from that first nitrotoluene metabolite i.e. toluene, produced by the H-I cells. This is achieved by making H-II cells mutants unable to transport and phosphorylate fructose i.e. PTSFRU gene knock-out.

Disclosed herein is a novel, cofactor balanced, cell-free biocatalysis pathway to make diacids from formaldehyde or methanol that does not use ATP and produces no CO.sub.2. In another embodiment, disclosed herein is a novel C5/C6 (hydrolysate) utilization pathway that interplays with the cell making diacids from formaldehyde or methanol that does not use ATP, produces no CO.sub.2, and replaces ATP with (cheap) polyphosphate (no other cofactors needed). Using methods and compositions disclosed herein, no CO.sub.2 is produced and the pathways disclosed herein are cell-free. The combination of pathways, feedstocks and products disclosed herein is novel.

A novel luciferase with a small molecular weight is provided. A polypeptide with luciferase activity comprising an amino acid sequence (A) or (B) is provided:

(A) an amino acid sequence as set forth in SEQ ID NO: 1 with deletion of amino acid residues at positions 1 to 69 and 204 to 221; or

(B) an amino acid sequence as set forth in SEQ ID NO: 1 with deletion of amino acid residues at positions 1 to 69 and deletion or substitution of at least one of amino acid residues 146 to 156.

Biomanufacturing hygiene control in batch and fed-batch bioreactors is provided using a bioreactor employing a chlorite/Cld system, wherein the bioreactor comprises an engineered cultured cell expressing a recombinant cytoplasmic chlorite dismutase (cCld) sufficient to increase chlorite resistance of the cell, and chlorite sufficient to substantially inhibit growth of one or more contaminating microorganisms in the bioreactor yet not substantially inhibit growth of the cell.

Provided herein are isolated polynucleotide encoding modified click beetle luciferase polypeptides that have enhanced luminescence and longer wavelength near-infrared signals. The disclosure also relates to near-infrared bioluminescence systems that include said modified click beetle luciferase polypeptides and novel luciferin derivatives, as well as methods of using said modified click beetle luciferase polypeptides and bioluminescence systems.

The subject invention provides novel plants that are not only resistant to 2,4-D and other phenoxy auxin herbicides, but also to aryloxyphenoxypropionate herbicides. Heretofore, there was no expectation or suggestion that a plant with both of these advantageous properties could be produced by the introduction of a single gene. The subject invention also includes plants that produce one or more enzymes of the subject invention alone or “stacked” together with another herbicide resistance gene, preferably a glyphosate resistance gene, so as to provide broader and more robust weed control, increased treatment flexibility, and improved herbicide resistance management options. More specifically, preferred enzymes and genes for use according to the subject invention are referred to herein as AAD (aryloxyalkanoate dioxygenase) genes and proteins. No α-ketoglutarate-dependent dioxygenase enzyme has previously been reported to have the ability to degrade herbicides of different chemical classes and modes of action. This highly novel discovery is the basis of significant herbicide tolerant crop trait opportunities as well as development of selectable marker technology. The subject invention also includes related methods of controlling weeds. The subject invention enables novel combinations of herbicides to be used in new ways. Furthermore, the subject invention provides novel methods of preventing the formation of, and controlling, weeds that are resistant (or naturally more tolerant) to one or more herbicides such as glyphosate.

The subject invention provides novel plants that are not only resistant to 2,4-D and other phenoxy auxin herbicides, but also to aryloxyphenoxypropionate herbicides. Heretofore, there was no expectation or suggestion that a plant with both of these advantageous properties could be produced by the introduction of a single gene. The subject invention also includes plants that produce one or more enzymes of the subject invention alone or “stacked” together with another herbicide resistance gene, preferably a glyphosate resistance gene, so as to provide broader and more robust weed control, increased treatment flexibility, and improved herbicide resistance management options. More specifically, preferred enzymes and genes for use according to the subject invention are referred to herein as AAD (aryloxyalkanoate dioxygenase) genes and proteins. No α-ketoglutarate-dependent dioxygenase enzyme has previously been reported to have the ability to degrade herbicides of different chemical classes and modes of action. This highly novel discovery is the basis of significant herbicide tolerant crop trait opportunities as well as development of selectable marker technology. The subject invention also includes related methods of controlling weeds. The subject invention enables novel combinations of herbicides to be used in new ways. Furthermore, the subject invention provides novel methods of preventing the formation of, and controlling, weeds that are resistant (or naturally more tolerant) to one or more herbicides such as glyphosate.