Abstract: The present disclosure describes Bst polymerase variants which exhibit DNA-dependent DNA polymerase and reverse transcriptase activity, and methods of use thereof. The Bst polymerase variants of the disclosure may be combined with a separate enzyme having reverse transcriptase activity to amplify target RNA sequences; however, the addition of a separate enzyme having reverse transcriptase activity is not necessary for the successful amplification of RNA targets using the Bst polymerase variants of the disclosure. Target DNA sequences may also be amplified using Bst polymerase variants of the disclosure.

Abstract: Disclosed herein are recombinant methods of activating expression of one or more biosynthetic gene clusters comprising more than one gene, the method comprising a recombinant DNA expression vector that possess two opposable inducible promoters that drives expression of a biosynthetic gene cluster exogenously from outside of the cluster to produce polyketides or non-ribosomal peptides in a heterologous host.

Abstract: Disclosed herein are recombinant methods of activating expression of one or more biosynthetic gene clusters comprising more than one gene, the method comprising a recombinant DNA expression vector that possess two opposable inducible promoters that drives expression of a biosynthetic gene cluster exogenously from outside of the cluster to produce polyketides or non-ribosomal peptides in a heterologous host.

Abstract: Disclosed herein are recombinant methods of activating expression of one or more biosynthetic gene clusters comprising more than one gene, the method comprising a recombinant DNA expression vector that possess two opposable inducible promoters that drives expression of a biosynthetic gene cluster exogenously from outside of the cluster to produce polyketides or non-ribosomal peptides in a heterologous host.

Abstract: A system and method for controlling a grapple is described in embodiments herein. A variable pressure control may be implemented that controls the pressure applied to a load held by the grapple. An interlock mechanism may be employed on the felling grapple that disables the operation of components of the felling grapple such as a feller. A grapple release mechanism may be activated to release the pressure applied to the load in the event of a malfunction in the felling grapple.

Abstract: An insulating felling grapple is used with an insulating utility vehicle. The utility worker stands on the ground near the worksite and remotely controls the insulating felling grapple and the utility vehicle to perform the task. In addition to other standard trimming and removal tasks, the insulating felling grapple is adapted to allow for the removal of energized vegetation material. In one method, the utility worker grasps the energized vegetation material, lifts or moves the energized vegetation material such that it is no longer energized, severs the non-energized vegetation material, and moves the severed vegetation material to a discard location. A boom assembly of the utility vehicle includes a primary insulation segment. The felling grapple includes a secondary insulation segment, as well as a grapple and a feller. The grapple has at least two tines for gripping the energized material. The feller is configured to sever the energized or nonenergized material.

Abstract: Linear vectors derived from bacteriophage of E. coli and host cells suitable for cloning are provided. The linear vectors include a left arm comprising a left telomere and a first selectable marker, a right arm comprising a right telomere and a second selectable marker and a cloning region located between the left arm and the right arm. Optional further components of the vector include transcriptional termination sequences, multiple cloning sites and reporter stuffer regions.

Abstract: Described herein are isolated polynucleotides that encode a fluorescent protein which is at least 80% sequence identical to a protein selected from the group consisting of SEQ. ID. NOS: 2, 4, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, and 74. Also described are expression constructs containing the polynucleotides, transformed host cells containing the expression constructions, the encoded fluorescent proteins themselves, and methods of using the nucleotides and encoded fluorescent proteins for bioanalytical research.

Abstract: Described herein are isolated polynucleotides that encode a fluorescent protein which is at least 80% sequence identical to a protein selected from the group consisting of SEQ. ID. NOS: 2, 4, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, and 74. Also described are expression constructs containing the polynucleotides, transformed host cells containing the expression constructions, the encoded fluorescent proteins themselves, and methods of using the nucleotides and encoded fluorescent proteins for bioanalytical research.

Abstract: Linear vectors derived from bacteriophage of E. coli and host cells suitable for cloning are provided. The linear vectors include a left arm comprising a left telomere and a first selectable marker, a right arm comprising a right telomere and a second selectable marker and a cloning region located between the left arm and the right arm. Optional further components of the vector include transcriptional termination sequences, multiple cloning sites and reporter stuffer regions.

Abstract: Described herein are isolated polynucleotides that encode a fluorescent protein which is at least 80% sequence identical to a protein selected from the group consisting of SEQ. ID. NOS: 2, 4, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, and 74. Also described are expression constructs containing the polynucleotides, transformed host cells containing the expression constructions, the encoded fluorescent proteins themselves, and methods of using the nucleotides and encoded fluorescent proteins for bioanalytical research.

Abstract: Linear vectors derived from bacteriophage of E. coli and host cells suitable for cloning are provided. The linear vectors include a left arm comprising a left telomere and a first selectable marker, a right arm comprising a right telomere and a second selectable marker and a cloning region located between the left arm and the right arm. Optional further components of the vector include transcriptional termination sequences, multiple cloning sites and reporter stuffer regions.

Abstract: A system for ligase-free cloning and/or expressing a target gene is described herein. A preferred version of the invention includes an E. coli host. The host preferably includes a T7 RNA polymerase gene comprising a T7gpl coding sequence, a lacUV5 promoter, and a lac operator. The host preferably further includes a lacI gene comprising a lacI coding sequence with an ATG start codon, a promoter derived from the lacql allele, and a translational enhancer derived from a 5? RNA leader sequence of T7 gene 10. The invention further includes a low-copy plasmid vector comprising a T7 promoter a lac operator operationally linked to the T7 promoter. The system is configured to inhibit target gene expression when uninduced and to permit gene expression upon induction by auto-induction.

Abstract: The present invention is directed to DNA polymerase fusion proteins with increased processivity and nucleic acid affinity. The invention includes a fusion protein comprising a nucleic acid-binding domain fused to a polymerase domain. The nucleic acid binding domain contains at least one nucleic acid binding motif, such as a DNA-binding motif or an RNA-binding motif. The nucleic acid binding domain preferably embodies an oligonucleotide/oligosaccharide binding (OB) fold, among other conformations. The invention further includes methods of synthesizing nucleic acids using the fusion proteins described herein.

Abstract: Thermostable viral and microbial polymerases exhibiting a combination of activities selected from proofreading (3?-5?) exonuclease activity, nick translating (5?-3?) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, terminal transferase activity, primase activity, and/or efficient incorporation of chain terminating analogs. Some of the polymerases provided herein include a first motif and a second motif. The first motif preferably has the sequence X1X2X3DX4PX5IELRX6X7X8, wherein X1 is I or V; X4 is F or Y; X8 is G or A; and X2, X3, X5, X6, and X7 are any amino acid. The second motif preferably has the sequence RX9X10X11KSANX12GX13X14YG, wherein X11 is G or A; X12 is F, L, or Y; X13 is L or V; X14 is I or L; and X9 and X10 are any amino acid.

Abstract: Thermostable viral polymerases exhibiting a combination of activities selected from, proofreading (3?-5?) exonuclease activity, nick translating (5?-3?) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, and/or decreased discrimination against incorporation of nucleotide analogs. Also provided are compositions including the polymerases, polynucleotides encoding the polymerases and methods of using the polymerases.

Abstract: A system for ligase-free cloning and/or expressing a target gene is described herein. A preferred version of the invention includes an E. coli host. The host preferably includes a T7 RNA polymerase gene comprising a T7gpl coding sequence, a lacUV5 promoter, and a lac operator. The host preferably further includes a lacI gene comprising a lacI coding sequence with an ATG start codon, a promoter derived from the lacql allele, and a translational enhancer derived from a 5? RNA leader sequence of T7 gene 10. The invention further includes a low-copy plasmid vector comprising a T7 promoter a lac operator operationally linked to the T7 promoter. The system is configured to inhibit target gene expression when uninduced and to permit gene expression upon induction by auto-induction.

Abstract: Vector preparations and cloning constructs suitable for use in cloning are provided. Vector preparations are double-stranded DNA molecules having two 3? termini, each terminus having a single base pair overhang that is capable of hybridizing to a single base pair overhang on a double stranded polynucleotide sequence to be cloned. The overhang of the vector preparation is suitably a dCMP and the overhang of the polynucleotide sequence to be cloned is suitably a dGMP. In other embodiments, the overhang of the polynucleotide sequence to be cloned is any ddNTP and the corresponding overhang of the vector preparation is any base that pairs to the ddNTP. The latter embodiment is particularly suited to preparing recombinant molecules having only a single insert. Methods of cloning, methods of preparing libraries of recombinant molecules and kits for carrying out the methods are also provided.

Abstract: Linear vectors derived from bacteriophage of E. coli and host cells suitable for cloning are provided. The linear vectors include a left arm comprising a left telomere and a first selectable marker, a right arm comprising a right telomere and a second selectable marker and a cloning region located between the left arm and the right arm. Optional further components of the vector include transcriptional termination sequences, multiple cloning sites and reporter stuffer regions.

Abstract: Thermostable viral polymerases exhibiting a combination of activities selected from, proofreading (3?-5?) exonuclease activity, nick translating (5?-3?) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, and/or decreased discrimination against incorporation of nucleotide analogs. Also provided are compositions including the polymerases, polynucleotides encoding the polymerases and methods of using the polymerases.