5470706 Process for the rescue of DNA and for detecting mutations in marker genes

PATENT ABSTRACTS

transmembrane glycoproteins might be important for the optimal structure of the virus and thus for its infectivity.

121

5470705 PROBE COMPOSITION CONTAINING A BINDING DOMAIN AND POLYMER CHAIN AND METHODS OF USE

5470703 METHOD

FOR

PEPTIDE

C-TERMINAL FRAGMENT SEQUENCE ANALYSIS AND APPARATUS COLLECTING

FOR PEPTIDE

FRAGMENT Nokihara Kiyosh; Yamamoto Rintar Kyoto, JAPAN assigned to Shimadzu Corporation The present invention relates to a method for peptide C-terminal fragment sequence analysis, in which the fragment collection is carried out on an allylamine group-derivatized polymer membrane or on allylamine group-derivatized glass fiber filter paper; the collected C-terminal fragment is immobilized thereon using a water-soluble carbodiimide etc.; and the obtained immobilized product is subjected directly to amino acid sequence analysis. The present invention also relates to an apparatus for collecting a peptide fragment. According to the method of the present invention, peptides which are rich in hydrophobic groups in their C-terminus and are therefore difftcult to trap with polyvalent ion carriers, currently used in the gas-phase sequencer, can be completely analyzed up to their C-terminus. Also, amino acid sequence analysis can be made even when the amount of C-terminal fragments is very small. In addition, since collection and immobilization of fragments can be done in the same bottle, the risks of contamination and mechanical loss are very low.

Grossman Paul; Fung Steven; Menchen Steven M; Woo Sam L; Winn-Deen Emily Burlingame, CA, UNITED STATES assigned to Applied Biosystems Inc Method and composition for detecting one or more selected polynucleotide regions in a target polynucleotide. In the method, a mixture of sequence-specific probes are reacted with the target polynucleotide under hybridization conditions, and the hybridized probes are treated to selectively modify those probes which are bound to the target polynucleotide in a base-specific manner. The resulting labeled probes include a polymer chain which imparts to each different-sequence probe, a distinctive ratio of charge/translational frictional drag, and a detectable label. The labeled probes are fractionated by electrophoresis in a non-sieving matrix, and the presence of one or more selected sequences in the target polynucleotide are detected according to the observed electrophoretic migration rates of the labeled probes in a non-sieving medium.

5470706 PROCESS

FOR

THE

RESCUE

OF DNA AND FOR DETECTING MUTATIONS IN MARKER GENES Vijg Ja; Gossen Jan A Rotterdam, NETHERLANDS assigned to Nederlandse Organisatie voor toegepastwetenschappelijk onderzoek TN0

122

PATENT ABSTRACTS

The invention relates to a process for the rescue and cloning of a DNA sequence wherein a vector containing said DNA sequence is multiplied in a suitable bacterial host. As the bacterial host, a bacterial strain which is incapable of host restriction is used, such as an Escherichia coli C strain. Preferably, a bacteriophage vector which is a suitable cloning vehicle for the bacterial host concerned is used as the vector, the step of in vitro packaging into phage coats being performed by means of a phage packaging extract obtained from a bacteria1 strain which is incapable of host restriction. The invention may be used in a process for detecting mutations in one or more marker genes introduced into a mammalian genome by means of a vector, which process comprises isolating DNA from cells of the mammal or from the transgenic mammalian cells, recovering the vector from said DNA, multiplying the vector in a suitable bacterial host which is deficient with respect to at least one of the marker genes, and testing marker gene expression. Preferably, the DNA isolated from cells of the transgenic mammal or the mammalian cells is pre-purified by fragmenting the DNA by means of a restriction enzyme which does not have a cutting site within the vector, separating the resulting fragments on the basis of a suitable criterion, such as differences in size, and collecting fragments comprising the vector, whereafter the vector is recovered from said pre-purified DNA.

5470707 HYDROGEN BOND LABELING AND BASE SEQUENCE DETERMINATION METHODS FOR DNA OR RNA Sasaki Yuj; Harada Yoshinor Kawagoe, JAPAN assigned to Hitachi Ltd

A hydrogen bond labeling and base sequence determination method for DNA or RNA takes advantage of the complementary nature of the fundamental structure of DNA or RNA bases to bind corresponding chemical species with the base species of a denatured DNA or RNA chain. After providing an aqueous solution containing a nucleic acid, heating the aqueous solution to cleave the nucleic acid; cooling the aqueous solution and adding a base-specific labeling molecule having available hydrogen bonds to the aqueous solution, the base-specific labeling molecules bond to the available bases in a one-to-one fashion via the available hydrogen bonds. The resulting labeled molecule can be microscopically observed to determine the base sequence of the nucleic acid.

5470708 PARTICLE-MEDIATED TRANSFORMATION OF MAMMALIAN UNATTACHED CELLS William; Yang Ning-Sun; Swain Burkholder Joseph K; Fuller Deborah L Verona, WI, UNITED STATES A method of genetically transforming mammalian unattached cells is disclosed. The method begins by preparing copies of a nucleic acid construct and coating these copies onto biologically inert carrier particles. Mammalian unattached cells are isolated in a liquid suspension. The cell suspension is placed on a target surface, wherein the liquid is spread to a thin film on the target surface. In an alternative embodiment of the present invention, the liquid is spread onto a porous surface. The with the cells are bombarded construct-coated particles in such a fashion