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The Rockefeller University

The Rockfeller University

Peter Model

Nathaniel Heintz

Xiangdong W Yang

Norman D Zinder

Jef D Boeke

Shiaoching Gong

A simple method for modifying genes in a recombination deficient host cell is disclosed. Such modifications include generating insertions, deletions, substitutions, and/or point mutations at any chosen site in the independent origin based cloning vector. The modified gene is contained in an independent origin based cloning vector that is used to introduce a modified heterologous gene into a cell. Such a modified vector may be used in the production of a germline transmitted transgenic animal, or in gene targeting protocols in eukaryotic cells. In particular, high throughput methodology is provided for generating the modified the independent origin based cloning vectors of the present invention.

Methods of performing homologous recombination based modification of nucleic acids in recombination deficient cells and use of the modified nucleic acid products thereof

A simple method for modifying genes in a recombination deficient host cell is disclosed. Such modifications include generating insertion, deletions, substitutions, and/or point mutations at any chosen site in the independent origin based cloning vector. The modified gene can be contained in an independent origin based cloning vector that is used to introduce a modified heterologous gene into a cell. Such a modified vector may be used in the production of a germline transmitted transgenic animal, or in gene targeting protocols in eukaryotic cells.

Methods of performing homologous recombination based modification of

A pallet and pallet foot is made of corrugated materials such that the entire pallet and pallet foot may be recycled with other corrugated materials (e.g., boxes or the like). The pallet foot is made from shredded corrugated waste which is then placed, along with an adhesive, in a mold under pressure and elevated temperature to form a molded pallet foot. The pallet foot may be glued to a corrugated deck provided with cutouts in the deck surface. Cutouts accept pallet feet from another pallet such that multiple pallets may be stacked in a nesting manner. The pallet feet have high resistance to dampness and may be further fortified with a water resistant coating. The resulting pallet is lightweight, strong, and readily recyclable.

Corrugated pallet and pallet foot

A pallet and pallet foot is made of corrugated materials such that the entire pallet and pallet foot may be recycled with other corrugated materials (e.g., boxes or the like). The pallet foot is made from shredded corrugated waste which is made into a slurry and molded, using a wet pulp process, to form a molded pallet foot. The pallet foot may be glued to a corrugated deck provided with cutouts in the deck surface. Cutouts accept pallet feet from another pallet such that multiple pallets may be stacked in a nesting manner. The pallet feet may be further fortified with a water resistant coating. The resulting pallet is lightweight, strong, and readily recyclable.

A method of externalizing proteins from the periplasmic space of gram-negative bacteria and in particular, E. coli and its relatives, comprising utilizing bacteria which have a phage gene, coding for a protein (such as gene III protein) or a mutant bacterial gene (such as a gene coding for a membrane function) which causes perturbation of the outer bacterial membrane resulting in leakage of proteins in the periplasmic space from the cell.

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New York, NY, United States of America

Peter Model