Described herein are synthetic methods for producing sequence-specific RNA oligonucleotides that eliminate impurities produced in prior art methods. In one aspect, an end-protected capture DNA complementary to a portion of the product RNA is employed. In another aspect, the template DNA is covalently or noncovalently linked to the RNA polymerase, either directly or through the use of a nontemplate DNA. In a third aspect, a flow chamber is employed. All of the methods can be used in combination.

The present invention generally relates to the microbiological industry, and specifically to the production of L-serine or L-serine derivatives using genetically modified bacteria. The present invention provides genetically modified microorganisms, such as bacteria, wherein the expression of genes encoding for enzymes involved in the degradation of L-serine is attenuated, such as by inactivation, which makes them particularly suitable for the production of L-serine at higher yield. The present invention also provides means by which the microorganism, and more particularly a bacterium, can be made tolerant towards higher concentrations of serine. The present invention also provides methods for the production of L-serine or L-serine derivative using such genetically modified microorganisms.

The invention relates to methods for template-independent synthesis of nucleic acids, comprising iteratively contacting an initiator sequence comprising a 3′-end nucleotide with a free 3′-hydroxyl group, with at least one nucleoside triphosphate, or a combination of nucleoside triphosphates, in the presence of an archaeal DNA primase or a functionally active fragment and/or variant thereof, thereby covalently binding said nucleoside triphosphate to the free 3-hydroxyl group of the 3-end nucleotide. It also relates to isolated functionally active fragments of archaeal DNA primases which are capable of template-independent terminal nucleotidyl transferase activity but are devoid of a template-independent primase activity.

Disclosed herein is a device for synthesizing ribonucleic acids (RNAs). According to embodiments of the present disclosure, the device comprises an in vitro transcription (IVT) module, a digestion module, and a processor. Optionally, the present device further comprises an IVT reaction monitoring means, a digestion reaction monitoring means, and/or a purifying means. Also disclosed herein are the methods of synthesizing RNA by use of the present device.

Described is a nucleic acid ligand, a mixture thereof, and the use thereof. The mixture contains two or more nucleic acid polymerase substrate analogs. The nucleic acid polymerase substrate analog is a single nucleic acid molecule or nucleic acid molecule analog which forms complementary pairing within a molecule, or a single or two nucleic acid molecules or nucleic acid molecule analogs which form complementary pairing between molecules; and a structure formed thereby has the characteristics of a nucleic acid polymerase substrate. The nucleic acid polymerase substrate analog is suitable for all polymerases and can be widely used in the field of nucleic acid amplification. The 3′ end of the nucleic acid ligand has a modification which inhibits the extension thereof.

The present invention provides methods for large-scale production of a composition enriched for full-length mRNA molecules using an SP6 RNA polymerase and compositions produced using such methods and uses thereof.

Insect inhibitory compositions comprising certain Chryseobacterium organisms, compositions and compounds derived from Chryseobacterium organisms, methods of using the compositions to inhibit insects that are injurious to humans, animals, and plants, and methods of making the compositions are disclosed.

A method for preparing nucleic acid sequences using an enzyme, including: (1) providing a reaction substrate having a pretreated surface. (2) Disposing a nucleotide having a terminal protecting group on the pretreated surface by a reaction enzyme, and a reaction temperature is 45° C.-105° C. (3) Removing the terminal protecting group of the nucleotide by irradiation or heating. (4) Coupling another nucleotide having the terminal protecting group to the nucleotide by the reaction enzyme, and a reaction temperature is 45° C.-105° C. (5) Determining whether nucleic acid sequence is completed, and if so, obtaining the nucleic acid sequence, if otherwise repeating steps (3) and (4). The method for preparing nucleic acid sequences using an enzyme of the invention may increase the efficiency of preparing nucleic acid sequences.

The present disclosure provides RNA polymerase variants for high efficiency transcription.

A protein expression system for use in a prokaryotic host is provided, the expression system comprising: a) an expression cassette comprising a nucleic acid sequence encoding a protein of interest operably linked to a T7 RNA polymerase-dependent promoter; and b) an expression cassette comprising a nucleic acid sequence encoding T7 RNA polymerase operably linked to a host polymerase-dependent λ phage promoter and a single perfect palindrome operator sequence; wherein the expression cassette for T7 RNA polymerase is located on the chromosome of a host cell.