Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Reexamination Certificate
1999-05-14
2001-09-18
Zitomer, Stephanie W. (Department: 1655)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
C435S006120, C435S069100, C435S091100, C435S252300, C536S023100, C536S025300
Reexamination Certificate
active
06291213
ABSTRACT:
BACKGROUND OF THE INVENTION
Throughout this application, various publications are referenced by author and date. Full citations for these publications may be found listed alphabetically at the end of the specification immediately preceding Sequence Listing and the claims. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein.
Attention of research scientists has recently focused upon various methods of creating libraries. The use of libraries is widespread in research and in the pharmaceutical industry. Libraries may consist of nucleic acid, peptides or even virtual molecules on a computer-readable material. Methods for the creation of libraries that are representative of the desired entity and that are useable has been a long felt challenge. In general, library construction has entailed many steps before the production of a final, useable library.
The following U.S. Patents are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to one of ordinary skill in the art. U.S. Pat. No. 5,498,530, Peptide Library and Screening Method; U.S. Pat. No. 5,491,074 Association Peptides; U.S. Pat. No. 5,432,018 Peptide Library and Screening Systems; U.S. Pat. No. 5,427,908 Recombinant Library Screening Methods; U.S. Pat. No. 5,338,665 Peptide Library and Screening Method; U.S. Pat. No. 5,270,170 Peptide Library and Screening Method; U.S. Pat. No. 5,541,061 Methods for Screening Factorial Chemical Libraries; U.S. Pat. No. 5,482,845, Method for Construction of Normalized cDNA Libraries; U.S. Pat. No. 5,512,463, Enzymatic inverse polymerase chain reaction library mutagenesis.
There are also peptide libraries and chimeric libraries that have been described. For example, see U.S. Pat. No. 5,525,486, Process for constructing cDNA library, and novel polypeptide and DNA coding for the same; U.S. Pat. No. 5,565,332, Production of chimeric antibodies—a combinatorial approach; U.S. Pat. No. 5,521,077, Method of Generating Multiple Protein Variants and Populations of Protein Variants Prepared thereby; U.S. Pat. No. 5,324,663, Methods and Products for the Synthesis of oligosaccharide structure on glycoproteins, glycolipids, or as free molecules, and for the isolation of cloned genetic sequences that determine these structures.
There have been combinatorial libraries also described which are usually composed of organic molecules attached to a solid support. A recent description of recently published patent applications may be found in Nature Biotechnology, Vol 14:1028-1029. Therein, the following published patent applications and patents were listed and described: Patent No. GB 2295152 A, solid phase synthesis of chemical library on flat solid support sheets divided into identifiable reaction zones; WO 9612014 A, repertoire of oligonucleotide tags comprise molecular tagging system used to track identify and sort molecules; WO 9607754, Oligonucleotides for inducing mutagenesis in an Ig light chain CDR; WO 9603424 A, combinatorial library comprising Diels-Alder products easily functionalized to form peptidomimetics for treating, e.g. Parkinson's disease; WO 9603418 A, Soluble combinatorial library by solid phase synthesis by using soluble polymeric support for core molecule attachment and buildup; WO 9603212 A, multidimensional device for synthesis of combinatorial chemical libraries comprising stacked trays of synthesis cells supplied with substrates and reagents.
SUMMARY OF THE INVENTION
The present invention provides for a method for generating a directed, recombinant fusion nucleic acid molecule which comprises: (a) contacting a first pair of single-stranded primers with a first strand and a second strand of a first nucleic acid molecule and a second pair of single-stranded primers with a first strand and a second strand of a second nucleic acid molecule under hybridization conditions, wherein the primers are suitable for use in a polymerase chain reaction, and (i) the first primer of the first pair of primers comprises a sequence that is homologous to the first strand of the first nucleic acid molecule; (ii) the second primer of the first pair of primers comprises a 3′ sequence that is homologous to the second strand of the first nucleic acid molecule and a 5′ sequence; (iii) the first primer of the second pair of primers comprises a 3′ sequence homologous to the first strand of the second nucleic acid molecule and a 5′ sequence that is complementary to the 5′ sequence of the second primer of the first pair of primers, and (iv) the second primer of the second pair of primers comprises a sequence that is homologous to the second strand of the second nucleic acid molecule; (b) amplifying the first nucleic acid molecule and the first pair of primers and the second nucleic acid molecule and the second pair of primers under amplification conditions, separately; (c) mixing the amplification products from step (b) and the first primer of the first pair of primers and the second primer of the second pair of primers under hybridization conditions; (d) amplifying the hybridized molecules of step (c) under amplification conditions so as to generate a directed, recombinant fusion nucleic acid molecule.
REFERENCES:
patent: 5942422 (1999-08-01), Rothstein
Jones et al. “DNA mutagenesis and recombination” Apr. 19, 1990, Nature, 344: 793-794.
Cooper & Dunham LLP
Forman B J
The Trustees of Columbia University in the City of New York
White John P.
Zitomer Stephanie W.
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