184results about "Polypeptide with MBP-tag" patented technology

A method for the ligation of expressed proteins which utilizes inteins, for example the RIR1 intein from Methanobacterium thermotrophicum, is provided. Constructs of the Mth RIR1 intein in which either the C-terminal asparagine or N-terminal cysteine of the intein are replaced with alanine enable the facile isolation of a protein with a specified N-terminal, for example, cysteine for use in the fusion of two or more expressed proteins. The method involves the steps of generating a C-terminal thioester-tagged target protein and a second target protein having a specified N-terminal via inteins, such as the modified Mth RIR1 intein, and ligating these proteins. A similar method for producing a cyclic or polymerized protein is provided. Modified inteins engineered to cleave at their C-terminus or N-terminus, respectively, and DNA and plasmids encoding these modified inteins are also provided.

The present invention relates to antibodies specific for a particular epitope on CD40 and antibodies that bind CD40 and have particular functional characteristics. The present invention also relates to fragments of these antibodies, uses of the antibodies for reduction or treatment of transplant rejection and graft-versus-host disease, and methods for making the antibodies.

Ligand-responsive chimeric proteins are engineered to cause a detectable output in response to a preselected stimulus. The engineered chimeric proteins are useful in industrial, commercial, medical, and scientific fields as a tool for programming a cellular response to a stimulus of choice and for use with in vitro assays. The engineered chimeric proteins include a detection domain and an interaction domain. Interaction of the engineered chimeric protein with a target biomolecule is modulated by the presence or absence of the preselected stimulus.

A self-cleaving element for use in bioseparations has been derived from a naturally occurring, 43 kDa protein splicing element (intein) through a combination of protein engineering and random mutagenesis. A mini-intein (18 kDa) previously engineered for reduced size had compromised activity and was therefore subjected to random mutagenesis and genetic selection. In one selection a mini-intein was isolated with restored splicing activity, while in another, a mutant was isolated with enhanced, pH-sensitive C-terminal cleavage activity. The enhanced cleavage mutant has utility in affinity fusion-based protein purification. The enhanced splicing mutant has utility in purification of proteins such as toxic proteins, for example, by inactivation with the intein in a specific region and controllable splicing. These mutants also provide new insights into the structural and functional roles of some conserved residues in protein splicing. Thus, disclosed and claimed are: a genetic system and self-cleaving inteins therefrom; bioseparations employing same; protein purification by inactivation with inteins in specific regions and controllable intein splicing; methods for determining critical, generalizable residues for varying intein activity; and products.

Peptides having at least 2 amino acid residues and no more than 15 amino acid residues are provided. The peptides comprise amino acid sequence X-Y or Y-X, wherein X is an aromatic amino acid and Y is any amino acid other than glycine. Also provided are pharmaceutical compositions and kits including such peptides as well as methods using same for diagnosing and treating amyloid associated diseases.

Provided are synthetic peptides based on a native sequence of HIV gp41 HR2 except that the synthetic peptides have a plurality of amino acid replacements comprising (a) a helix-promoting amino acid, or (b) a combination of helix-promoting amino acids, and charged amino acids introduced to form ion pairs in the synthetic peptide; wherein the synthetic peptides demonstrate an unexpected, improved biological activity, as compared to a peptide having an amino acid sequence without the plurality of amino acid substitutions. Also provided are polynucleotides encoding synthetic peptide, and methods of using these synthetic peptides in inhibition of, or as compositions to inhibit, transmission of HIV to a target cell.

A simple and efficient SUMO fusion protein expression system for producing native proteins.

Zinc finger proteins of the Cys₂His₂ type represent a class of malleable DNA binding proteins which may be selected to bind diverse sequences. Typically, zinc finger proteins containing three zinc finger domains, like the murine transcription factor Zif268 and the human transcription factor Spl, bind nine contiguous base pairs (bp). To create a class of proteins which would be generally applicable to target unique sites within complex genomes, the present invention provides a polypeptide linker that fuses two three-finger proteins. Two six-fingered proteins were created and demonstrated to bind 18 contiguous bp of DNA in a sequence specific fashion. Expression of these proteins as fusions to activation or repression domains allows transcription to be specifically up or down modulated within cells. Polydactyl zinc finger proteins are broadly applicable as genome-specific transcriptional switches in gene therapy strategies and the development of novel transgenic plants and animals. Such proteins are useful for inhibiting, activating or enhancing gene expression from a zinc finger-nucleotide binding motif containing promoter or other transcriptional control element, as well as a structural gene or RNA sequence.

The invention belongs to the fields of biotechnologies and genetic engineering technologies and in particular relates to an efficient soluble expression and purification method of Abeta42 in escherichia coli. An Abeta42 expression system constructed by the invention provides a construction method of an expression vector of prokaryote for soluble fusion expression of human amyloid protein Abeta42 and the expression vector is introduced into an escherichia coli expression host; target Abeta42 with high-purity bioactivity is obtained through induced expression, protein enzyme digestion, two-step purification and identification. According to the method provided by the invention, an escherichia coli expression system and fusion protein have the characteristics of high expression efficiency, large expression amount, low cost, easiness for operation and the like; an expressed Abeta42 polypeptide has the advantages of no residues, high purity, high productivity and the like.

The invention discloses a mannanase and the coding gene, a recombinant vector and a host cell comprising the gene, and expression of the recombinant vector in E.coli and production of the enzyme in yeast cells by using the recombinant vector. The present invention also provides a new wild type subspecies of Bacillus Megaterium, in particular, Bacillus Megaterium FB101.

The present invention relates to a new isolated polypeptide comprising an amino acid sequence having at least 94%, 95%, 99% or 100% identity to the full length amino acid sequence set forth in SEQ ID NO: 1, and having a polyester degrading activity, and uses thereof.

The present invention provides for methods and compositions for treating or preventing arthritis and joint injury.

A method for the ligation of expressed proteins which utilizes inteins, for example the RIR1 intein from Methanobacterium thermotrophicum , is provided. Constructs of the Mth RIR1 intein in which either the C-terminal asparagine or N-terminal cysteine of the intein are replaced with alanine enable the facile isolation of a protein with a specified N-terminal, for example, cysteine for use in the fusion of two or more expressed proteins. The method involves the steps of generating a C-terminal thioester-tagged target protein and a second target protein having a specified N-terminal via inteins, such as the modified Mth RIR1 intein, and ligating these proteins. A similar method for producing a cyclic or polymerized protein is provided. Modified inteins engineered to cleave at their C-terminus or N-terminus, respectively, and DNA and plasmids encoding these modified inteins are also provided.

Stabilized reverse transcriptase fusion proteins including a thermostable reverse transcriptase connected to a stabilizer protein are described. Attaching the stabilizer protein to the thermostable reverse transcriptase stabilizes the fusion protein and can aid in its purification, provide increased solubility, allow for longer storage, or allow the fusion protein to be used under more rigorous conditions such as higher temperature. The stabilized reverse transcriptase fusion protein can also include a linker between the stabilizer protein and the thermostable reverse transcriptase. The stabilized reverse transcriptase fusion proteins are suitable for use in nucleic acid amplification methods such as the reverse transcription polymerase chain reaction and other applications involving cDNA synthesis.

The present invention relates to variant endoglucanases having improved thermoactivity, improved thermostability, and improved viscosity reduction activity over wild-type M. thermophila endoglucanase.

A new SUMO-specific affinity tag is described herein, based on the amino acid sequence from 403-621 of the SUMO protease Ulp1, and containing a crucial substitution of serine for cysteine. This affinity tag is particularly useful for a range of applications, including detection and affinity purification of sumoylated proteins from cell extracts.