University of California

The Scripps Research Institute

Other

Agilent Technologies, Inc.

Disney Enterprises, Inc.

John Christopher Anderson

Lei Wang

Peter G Schultz

David R Liu

Thomas J Magliery

Eric L Meggers

Ryan Aaron Mehl

Miro Pastrnak

Zhiwen Zhang

Jason W Chin

Stephen William Santoro

Jason William Chin

Steven William Santoro

Bo U Curry

Andrew Witkin

David Sukovich

Benjamin Ernest Anderson

Markus De Raad

Cameron Davies

Miguel Campo-Rembado

Anjali Dange

Timothy Hsiau

Cyrus Modavi

Gabriel Lopez

Miro Pastranak

Peter Schultz

Synthetic auxotrophs with one or more ligand-dependent essential gene functions and methods of production that can be used for biosafety. The ligand-dependent function of an essential gene product can be produced by a series of mutations in the ORF of an essential gene; N, C, or insertional fusions of ligand-binding domains with essential genes or an engineered ligand-dependent intein splicing to alter essential gene function. A positive and/or negative selection can be used to identify auxotrophs from created mutant libraries. The positive selection is performed by growing a mutant library in conditions where growth or viability depends on the function of mutagenized essential genes. The negative selection eliminates constitutively growing cells that do not require a ligand for growth by growing the library in the absence of complementing ligand and in conditions where growing cells are eliminated. Desirable phenotypes are collected after the selections.

Synthetic auxotrophs with ligand dependent essential genes for biosafety

Traditional enzyme characterization methods are low-throughput, and therefore limit engineering efforts in synthetic biology and biotechnology. Here we propose a DNA-linked enzyme-coupled assay (DLEnCA) to monitor enzyme reactions in a high-throughput manner. Throughput is improved by removing the need for protein purification and by limiting the need for liquid chromatography mass spectrometry (LCMS) product detection by linking enzymatic function to DNA modification. DLEnCA is generalizable for many enzymatic reactions, and here we adapt it for glucosyltransferases, methyltransferases, and oxidoreductases. The assay utilizes cell free transcription/translation systems to produce enzymes of interest, while UDP-Glucose and T4-&#x3b2;-glucosyltransferase are used to modify DNA, which is detected post-reaction using qPCR or similar means of DNA analysis. For monitoring methyltransferases, consumption of SAM is observed by coupling to EcoRI methyltransferase. For monitoring oxidoreductases, consumption of NADH is observed by coupling to Taq orDNA ligase. OleD and two glucosyltransferases fromwere used to verify the assay's generality toward glucosyltransferases. Two methyltransferases from human andwere used to verify the assay's generality towards methyltransferases. We show DLEnCA's utility by mapping out the substrate specificity for these enzymes and observing the multiple steps of a biosynthetic pathway.

DNA-linked enzyme-coupled assays

The present invention provides methods for generating a library of synthetic polynucleotides. The present invention also provides methods for generating proteins encoded by the library of synthetic polynucleotides. In addition, provided herein are methods for determining the soluble expression of said proteins. This invention is based, in part, on the discovery of a method for selecting optimal oligonucleotides in combination with performing a phosphorylation reaction, ligation reaction and PCR amplification in a single reaction vessel to produce synthetic polynucleotides in a multiplex manner.

One-pot multiplex gene synthesis

In a networked system, a method and apparatus for measuring advertising effectiveness involves broadcasting an advertisement at a broadcast time, monitoring at least one social network or other online forum following the broadcast time, filtering social network traffic to identify messages, search engine queries to identify queries, or other traffic entries, that appear to be responsive to the broadcast advertisement, determining a time of initiation for at least a plurality of entries that appear to be responsive to the broadcast advertisement, and determining at least one demographic for at least the plurality of the entries that appear to be responsive to the broadcast advertisement.

Data mining to determine online user responses to broadcast messages

This invention provides compositions and methods for generating components of protein biosynthetic machinery including orthogonal tRNAs, orthogonal aminoacyl-tRNA synthetases, and orthogonal pairs of tRNAs/synthetases. Methods for identifying orthogonal pairs are also provided. These components can be used to incorporate unnatural amino acids into proteins in vivo.

Methods and compositions for the production of orthogonal tRNA-aminoacyl tRNA synthetase pairs

An enclosure which provides a storage of mobile phone headset cables as well as a means to extend and retract said headset cables is disclosed. The enclosure allows the headset cables to be wound internally onto a large diameter reel while a mobile phone connecting cable is wound onto a smaller spool, thereby allowing a greater length of headset cables to be extended. A button enables said cables to be retracted to eliminate slack while the device is in use, or to completely retract the cables when the device is disconnected from the phone. The retracted cable portions may be contained within the enclosure which further comprises a plurality of nests along external surfaces for the storage of ear buds, a microphone, and the phone connection plug in a flush manner, thereby avoiding the entanglement of exposed loose cables.

Mobile phone headset recoil device

Methods and composition for the production of orthogonal tRNA-aminoacyltRNA synthetase pairs

The invention provides methods and compositions for in vivo incorporation of unnatural amino acids. Also provided are compositions including proteins with unnatural amino acids.

In vivo incorporation of unnatural amino acids

Methods and composition for the production of orthogonal tRNA-aminoacyl tRNA synthetase pairs

Methods and compositions for the production of orthogonal tRNA-aminoacyl-tRNA synthetase pairs

Growing community of inventors

Berkeley, CA, United States of America

John Christopher Anderson