American Home Products Corporation

Apollon, Inc.

Alnylam Pharmaceuticals, Inc.

C Satishchandran

Catherine Julia Pachuk

Liat Mintz

Vincent R Zurawski, Jr

Vincent R Zurawski

Richard B Ciccarelli

Manoj Samuel

Linda Anne Snyder

Composition comprising lamellar vesicles that comprise a local anesthetic and a nucleic acid molecule are disclosed. Methods of making such compositions are disclosed. Method of delivering proteins to cells of individuals are disclosed. Methods of inducing immune responses in individuals are disclosed. Methods of delivering nucleic acid molecules to cells of individuals are disclosed.

Vesicular complexes and methods of making and using the same

Methods of detecting the presence of a plasmid DNA sequence integrated in a chromosomal DNA molecule of a eukaryotic cell in a sample that contains chromosomal DNA molecules of eukaryotic cells and free plasmid DNA molecules are disclosed. According to the invention, chromosomal DNA of eukaryotic cells which are free of deoxyadenosine methyltransferase, and free plasmid DNA molecules which are produced in cells that contain deoxyadenosine methyltransferase and which have a DpnI site, are digested with one or more restriction enzymes that cleave plasmid DNA sequences integrated in the chromosomal DNA and plasmid DNA molecules to produce DNA digestion segments that are then fractionated to produce a plurality of fractions. The DNA digestion segments in each fraction is digested with DpnI and plasmid DNA sequences are amplified using sets of primers that flank a DpnI site in the plasmid DNA sequence. The presence of amplified fragments indicates the presence of a plasmid DNA sequence integrated into the chromosomal DNA.

Method of detecting foreign DNA integrated in eukaryotic chromosomes

Chain reaction cloning methods and reagents and kits for performing such methods are provided. Chain reaction cloning allows ligation of double-stranded DNA molecules by DNA ligases and bridging oligonucleotides. Double-stranded nucleic acid molecules are denatured into single-stranded molecules. The ends of the molecules are brought together by hybridization to a template. The template ensures that the two single-stranded nucleic acid molecules are aligned correctly. DNA ligase joins the two nucleic acid molecules into a single, larger, composite nucleic acid molecule. The nucleic acid molecules are subsequently denatured so that the composite molecule formed by the ligated nucleic acid molecules and the template cease to hybridize to each. Each composite molecule then serves as a template for orienting unligated, single-stranded nucleic acid molecules. After several cycles, composite nucleic acid molecules are generated from smaller nucleic acid molecules. A number of applications are disclosed for chain reaction cloning including site-specific ligation of DNA fragments generated by restriction enzyme digestion, DNAse digestion, chemical cleavage, enzymatic or chemical synthesis, and PCR amplification.

Chain reaction cloning using a bridging oligonucleotide and DNA ligase

Methods of detecting the presence of linear DNA in a sample that contains circular DNA are disclosed. The methods comprise combining an ATP-dependent deoxyribonuclease, ATP and a sample to form a reaction mixture, maintaining the reaction mixture under conditions in which linear DNA is processed by the ATP-dependent deoxyribonuclease, and detecting the presence of ADP thus produced. Methods of detecting the presence of DNA, RNA, or protein in samples are disclosed. The methods comprise combining an ATP-dependent deoxyribonuclease, an ATP-dependent ribonuclease, or an ATP-dependent protease, respectively, with ATP and a sample to form a reaction mixture. The reaction mixture is maintained under conditions in which DNA, RNA or protein is processed by the respective ATP-dependent enzyme. The presence of ADP thus produced is detected, indicating the presence of the enzyme substrate. Methods of quantifying the DNA molecules, RNA molecules or protein molecules are also disclosed.

Method of detecting the presence and measuring the quantity of

Chimeric kanamycin resistance genes are disclosed. The chimeric genes comprise a nucleotide sequence that encodes ANT(4')-IA enzyme operably linked to a heterologous promoter and a heterologous termination sequence. Plasmids that comprise the chimeric kanamycin resistance gene are disclosed. Bacterial cells that comprise the chimeric gene on a plasmid or integrated into the bacterial genome are disclosed. Methods of producing plasmids are disclosed. Pharmaceutical compositions comprising plasmids that include the chimeric genes are disclosed. Methods of enhancing growth of a bacterial cells are disclosed. Plasmid which comprise the chimeric kanamycin resistance gene and the sequences from herpes simplex virus gene HSVgD.sub.2 or human immunodeficiency virus are disclosed.

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Lansdale, PA, United States of America

C Satishchandran