A recombinant vector of a thermolabile UNG fused protein and an expressing and purifying method are provided. The method comprises cloning a Cod UNG genetic sequence to a pCold-SUMO vector to construct a recombinant vector pCold-SUMO-Cod UNG, transforming to E. coli BL21 (DE3) competent cells, transforming and expressing molecular chaperone plasmids pG-Tf2, and inducing the expression at a low temperature to obtain a soluble SUMO-Cod UNG fused protein.

The present invention provides a method of modifying a targeted site of a double stranded DNA, including a step of contacting a complex wherein a nucleic acid sequence-recognizing module that specifically binds to a target nucleotide sequence in a selected double stranded DNA and DNA glycosylase with sufficiently low reactivity with a DNA having an unrelaxed double helix structure (unrelaxed DNA) are bonded, with the double stranded DNA, to convert one or more nucleotides in the targeted site to other one or more nucleotides or delete one or more nucleotides, or insert one or more nucleotides into the targeted site, without cleaving at least one strand of the double stranded DNA in the targeted site.

The present application relates to: a single base substitution protein; a composition comprising same; and a use thereof.

Materials and methods for using modified Cas9-APOBEC fusion polypeptides for targeted modification of specific DNA sequences are provided herein.

A method for nucleic acid synthesis and regeneration of a reusable synthesis initiator includes incorporating a linking nucleotide to an immobilized initiator using a polymerase, synthesizing a nucleic acid right after the linking nucleotide using the polymerase, subjecting a substrate base of the linking nucleotide in the nucleic acid to base-excision by a DNA glycosylase to generate an abasic site, subjecting the abasic site to cleavage by an endonuclease to release the nucleic acid from the initiator, and converting the 3′ terminus of the initiator back to its original form by a 3′ phosphatase activity-possessing enzyme. A kit based on the aforesaid method and a method for regenerating a reusable initiator are also disclosed.

Sequential assembly of oligonucleotide hairpins is used to create oligonucleotides with specific sequences. Each oligonucleotide hairpin includes a payload region containing one or more nucleotides that are added to the end of an anchor strand. Overhang regions on the oligonucleotide hairpins hybridize to anchor strands attached to a substrate. The hybridized oligonucleotide hairpins are covalently attached to the anchor strands by the activity of ligase. The oligonucleotide hairpins include an enzyme cleavage region which, when cleaved, separates the payload region from the remainder of the oligonucleotide hairpin. The oligonucleotide hairpin is separated from the anchor strand by denaturation and washed away. This process is repeated with additional oligonucleotide hairpins to add additional nucleotides to the ends of the anchor strands. A microelectrode array may be used to control the location of hybridization and create multiple oligonucleotides in parallel. Fully assembled oligonucleotides can be separated from the substrate and stored.

The subject invention provides microbe-based products, as well as their use in simultaneously enhancing oil recovery from an oil well while efficiently removing contaminating compositions such as biofilm, scale, paraffin, and/or asphaltenes from oil production equipment and oil-bearing formations. The subject invention can also be used to disperse paraffin and asphaltene precipitates, and to reduce the viscosity of heavy crude oil. The subject invention further provides materials and methods for bioremediation of hydrocarbon-contaminated sites.

The present disclosure provides nucleobase editors that include a cytidine deaminase domain, a codon-optimized nuclease-defective Cas9 domain, and at least one nuclear-localization sequence. The nucleobase editors disclosed herein improve the efficiency by which single-nucleotide variants can be created compared to conventional BE3 nucleobase editors.

The invention provides compositions and methods for assembling a DNA molecule having a desired sequence. The methods involve contacting a DNA polymerase, dNTPs, and a plurality of pairs of oligonucleotides. The oligonucleotides of a pair have a portion of the desired sequence, and an internal sequence that overlaps and is complementary to an internal sequence of the other oligonucleotide of the pair, and, when arranged in order, they have at least a portion of the desired sequence. The oligonucleotides also have a 3′ or a 5′ primer binding sequence having a binding site for a primer. The oligonucleotides that correspond to the end oligonucleotides of the desired sequence also have a universal 3′ flanking sequence and a universal 5′ flanking sequence, respectively.

The present disclosure provides for endonuclease enzymes having distinguishing domain features, as well as methods of using such enzymes or variants thereof.