Abstract: This disclosure provides several methods to generate nucleic acid mutations in vivo, for instance in such a way that no heterologous sequence is retained after the mutagenesis is complete. The methods employ integrative recombinant oligonucleotides (IROs). Specific examples of the described mutagenesis methods enable site-specific point mutations, deletions, and insertions. Also provided are methods that enable multiple rounds of mutation and random mutagenesis in a localized region. The described methods are applicable to any organism that has a homologous recombination system.

Abstract: The present invention provides isolated polypeptides of human p53 that contain mutations. These mutations can be toxic mutations, supertransactivating mutations or tox-suppressor mutations. Further provided by the invention are methods of identifying toxic, supertransactivating, weak transactivating and tox-suppressor mutations as well as methods of identifying compounds that mimic the toxic, supertransactivating and tox-suppressor mutations in human p53. Also provided are methods of inducing toxicity in a cell by administering a polypeptide comprising a supertransactivating or a toxic mutation.

Abstract: This disclosure provides several methods to generate nucleic acid mutations in vivo, for instance in such a way that no heterologous sequence is retained after the mutagenesis is complete. The methods employ integrative recombinant oligonucleotides (IROs). Specific examples of the described mutagenesis methods enable site-specific point mutations, deletions, and insertions. Also provided are methods that enable multiple rounds of mutation and random mutagenesis in a localized region. The described methods are applicable to any organism that has a homologous recombination system.

Abstract: The invention is directed to a method of making a yeast artificial chromosome (YAC) comprising introducing into yeast cells a population of nucleic acids and a vector, wherein the vector comprises a yeast centromere, a selectable marker, a yeast telomere, and a sequence which can recombine with a region of a nucleic acid within the population of nucleic acids, whereby in vivo recombination makes the YAC. The invention is also directed to a method of making a YAC using two vectors and a method of making a circular YAC. The invention is also directed toward methods of making YACs with a selected nucleic acid insert from a mixed population of nucleic acids using transformation-associated recombination.

Abstract: The present invention relates, in general, to endo-exonucleases. In particular, the present invention relates to DNA segments encoding for a polypeptide having an amino acid sequence corresponding to mammalian endo-exonuclease, a polypeptide having an amino acid sequence corresponding to mammalian endo-exonuclease, antibodies to mammalian endo-exonuclease, a recombinant DNA molecule, mutant cells substantially lacking endo-exonuclease activity, a cell containing mammalian endo-exonuclease, and methods of producing and using the polypeptide, DNA segment and mutants.

Abstract: A sensitive, yeast-based genetic system for identifying agents causing double-strand DNA damage is described. The system comprises a yeast strain containing either chromosomes having divergent (homeologous) DNA sequences, a single nonhomologous chromosome or a single artificial chromosome with suitable genetic markers so that double-strand damage leading to the loss of such chromosome due to the inability to undergo recombinational repair with a homolog is detected.

Abstract: A method of producing plasmids with heteroduplex DNA sequences, in which probe plasmids containing DNA inserts that correspond to probe sequences and test plasmids containing DNA inserts that correspond to a population of test sequences are first constructed. The vector portions of the plasmids used in these constructions have similar sequences. When test and probe plasmids are cleaved with appropriate restriction enzymes and then denatured to separate strands, complementary regions of the linear strands which correspond to vector sequences can anneal. The molecules that harbor test and probe inserts that are related by sequence complementarity form non-covalently closed circular molecules. These molecules can replicate after transformation into an appropriate host organism. There is a replication bias against plasmids which anneal through vector sequences, but which do not contain homologous probe and test inserts.

Abstract: The present invention relates, in general, to a DNA segment. In particular, the present invention relates to a DNA segment coding for a polypeptide having an amino acid sequence corresponding to RhoNUC, a polypeptide having an amino acid sequence corresponding to RhoNUC, antibodies to RhoNUC, a recombinant DNA molecule, a cell containing the recombinant DNA molecule, and methods of producing and using the polypeptide and DNA segment.