Abstract: Recombinant bacterial triple-helical collagen-like proteins comprising two or more repetitive sequences of Gly-Xaa-Yaa yielding high-stability polymeric constructs without the need for post-translational modifications and which may incorporate one or more functional domains of biological or structural importance.

Abstract: A method for screening for an antiviral agent capable of blocking a viral viroporin by determining whether a test agent can rescue expression of a fragment of a viral viroporin in a Single Protein Production system of Escherichia coli is provided.

Abstract: The present invention is directed to the improved methods for the temporal induction of proteins using the condensed single protein production (cSPP) system.

Abstract: Disclosed in certain embodiments is a method of inhibiting cell function comprising inducing the expression of a mRNA interferase that cleaves mRNA at GCU.

Abstract: The present invention describes a single-protein production (SPP) system in living E. coli cells that exploits the unique properties of an mRNA interferase, for example, MazF, a bacterial toxin that is a single stranded RNA- and ACA-specific endoribonuclease, which efficiently and selectively degrades all cellular mRNAs in vivo, resulting in a precipitous drop in total protein synthesis. Concomitant expression of MazF and a target gene engineered to encode an ACA-less mRNA results in sustained and high-level (up to 90%) target expression in the virtual absence of background cellular protein synthesis. Remarkably, target synthesis continues for at least 4 days, indicating that cells retain transcriptional and translational competence despite their growth arrest. SPP technology works well for yeast and human proteins, even a bacterial integral membrane protein.

Abstract: The present invention provides cold shock protein-containing compositions for improved DNA synthesis reactions with improved reactivity, methods for synthesizing DNA using such compositions, kits for use in such methods, and DNA compositions yielded by such methods. The present invention further provides cold shock protein-containing compositions for the identification of endoribonuclease cleavage sites, methods for identifying endoribonuclease cleavage sites using such compositions, and kits for use in such methods.

Abstract: The invention provides methods for identifying an agent which prevents or partially prevents an antitoxin from forming a complex with its cognate toxin, comprising contacting a potential agent with a labeled substrate in solution, whereby detection of the label indicates presence of an agent that prevents an antitoxin from forming complex with a toxin. The invention also provides agents capable of interfering with formation of a toxin-antitoxin complex. Such agents act as novel, non-conventional antibiotics against human pathogenic bacteria.

Abstract: Disclosed herein is a set of E. coli single-protein production (SPP) technologies with protein NMR (SPP-NMR) for (i) using isotope-enriched membrane proteins produced with the SPP system in screening detergent conditions suitable for purification and/or three-dimensional structure analysis without the requirement for protein purification, (ii) producing 2H, 13C, 15N enriched proteins suitable for high throughput and membrane protein NMR studies, and (iii) labeling with 13C—15N specific peptide bonds in proteins (referred to herein as SPP-PBL).

Abstract: Disclosed are methods for purifying toxin proteins which avoid denaturation and renaturation procedures.

Abstract: A deployment of a toxin gene for developmental programmed cell death in bacteria is described. M. xanthus is demonstrated to have a solitary mazF gene that lacks a cotranscribed antitoxin gene. Deletion of mazF results in elimination of the obligatory cell death during development causing dramatic reduction in spore formation. Surprisingly, MrpC functions as a MazF antitoxin and a mazF transcription activator. Transcription of mrpC and mazF is negatively regulated via MrpC phosphorylation by a Ser/Thr kinase cascade. Various methods of exploiting this novel pathway are described herein.

Abstract: The present invention describes a single-protein production (SPP) system in living E. coli cells that exploits the unique properties of an mRNA interferase, for example, MazF, a bacterial toxin that is a single stranded RNA- and ACA-specific endoribonuclease, which efficiently and selectively degrades all cellular mRNAs in vivo, resulting in a precipitous drop in total protein synthesis. Concomitant expression of MazF and a target gene engineered to encode an ACA-less mRNA results in sustained and high-level (up to 90%) target expression in the virtual absence of background cellular protein synthesis. Remarkably, target synthesis continues for at least 4 days, indicating that cells retain transcriptional and translational competence despite their growth arrest. SPP technology works well for yeast and human proteins, even a bacterial integral membrane protein.

Abstract: The present invention relates to a DNA molecule or vector and a host cell containing this DNA molecule or vector which can be used to produce a heterologous polypeptide under conditions that elicit a cold shock response in the host cell. The DNA molecule and vector include a nucleotide sequence encoding a heterologous polypeptide and a promoter and 5?-UTR from a cold shock inducible gene which directs its expression. In addition, an AT-rich sequence that enhances translation under cold shock inducible conditions is either present in the coding sequence of the heterologous polypeptide or in an additional element inserted between the coding sequence and the cold shock inducible promoter and 5?-UTR.

Abstract: The invention provides vectors containing a multiple cloning site comprising a PrS tag or a PrS2 tag from Myxococcus xanthus. Methods are provided for enhancing solubility of a target protein using Protein S tagged target fusion proteins.

Abstract: Ribonucleases, antibiotics, bacterial toxins and viruses inhibit protein synthesis, which results in apoptosis in mammalian cells. How the BCL-2 family of proteins regulates apoptosis in response to shutoff of protein synthesis is not known. According to the present invention, an Escherichia coli toxin MazF inhibited protein synthesis by cleavage of cellular mRNA, and induced apoptosis in mammalian cells. MazF-induced apoptosis required proapoptotic BAK and its upstream regulator, the proapoptotic BH3-only protein NBK/BIK, but not BIM, PUMA or NOXA. Furthermore, NBK/BIK- or BAK-deficient cells were resistant to cell death induced by pharmacologic inhibition of translation and by virus-mediated shutoff of protein synthesis. Thus, the BH3-only protein NBK/BIK is the apical regulator of a BAK-dependent apoptotic pathway in response to shutoff of protein synthesis.

Abstract: The invention relates to a gene isolated from E. coli, dep, which confers resistance to the antibacterial activity of 4,5-dihydroxy-2-cyclopenten-1-one (DHCP). The invention further relates to the putative protein encoded by dep, which is a hydrophobic, transmembrane efflux protein specific for DHCP. The invention further relates to plasmids containing the dep gene, and to bacterial cells expressing dep. Furthermore, the invention provides applications for use in conferring resistance to antibacterial activity in organisms. The dep gene can be used to identify compounds which inhibit the efflux activity responsible for the resistance to DHCP or to compounds which are functionally equivalent to DHCP.

Abstract: Regulatory elements of the 5?UTR of cold-shock inducible genes that prolong the expression of cold-shock inducible genes under conditions that elicit the cold-shock response in a bacterium, repress the expression of the cold-shock inducible genes under physiological conditions, or enhance the translation of cold-shock inducible genes under conditions that elicit a cold-shock response in bacteria; vectors incorporating these elements; and methods for expressing proteins.

Abstract: The present invention is directed to the discovery of a novel family of enzymes designated herein as mRNA interferases that exhibit endoribonuclease activity. The novel finding of the present inventors, therefore, presents new applications for which mRNA interferase nucleic and amino acid sequences, and compositions thereof may be used to advantage. The invention also encompasses screening methods to identify compounds/agents capable of modulating mRNA interferase activity and methods for using such compounds/agents. Also provided is a kit comprising mRNA interferase nucleic and/or amino acid sequences, mRNA interferase activity compatible buffers, and instruction materials.

Abstract: The invention relates to a gene, dep, which confers resistance to the antibacterial activity of 4,5-dihydroxy-2-cyclopenten-1-one (DHCP). The invention further relates to the putative protein encoded by dep, which is a hydrophobic, transmembrane efflux protein specific for DHCP. The invention further relates to plasmids containing the dep gene, and to bacterial cells expressing dep. Furthermore, the invention provides applications for use in conferring resistance to antibacterial activity in organisms. The dep gene can be used to identify compounds which inhibit the efflux activity responsible for the resistance to DHCP or to compounds which are functionally equivalent to DHCP.

Abstract: The present invention provides the multifunctional biological and biochemical sensor technology based on ZnO nanostructures. The ZnO nanotips serve as strong DNA or protein molecule binding sites to enhance the immobilization. Patterned ZnO nanotips are used to provide conductivity-based biosensors. Patterned ZnO nanotips are also used as the gate for field-effect transistor (FET) type sensors. Patterned ZnO nanotips are integrated with SAW or BAW based biosensors. These ZnO nanotip based devices operate in multimodal operation combining electrical, acoustic and optical sensing mechanisms. The multifunctional biosensors can be arrayed and combined into one biochip, which will enhance the sensitivity and accuracy of biological and biochemical detection due to strong immobilization and multimodal operation capability. Such biological and biochemical sensor technology are useful in detection of RNA-DNA, DNA-DNA, protein-protein, protein-DNA and protein-small molecules interaction.

Abstract: The present invention relates to a DNA molecule or vector and a host cell containing the DNA molecule or vector which can be used to produce a heterologous polypeptide under conditions that elicit a cold shock response in the host cell. The DNA molecule and vector include a nucleotide sequence encoding a heterologous polypeptide and a promoter and 5?-UTR from a cold shock inducible gene which directs its expression. IN addition, an AT-rich sequence that enhances translation under cold shock inducible conditions is either present in the coding sequence of the heterologous polypeptide or in an additional element inserted between the coding sequence and the cold shock inducible promoter and 5?-UTR.