4766062 displacement polynucleotide assay method and polynucleotide complex reagent therefor

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PATENT ABSTRACTS 63 4766062 DISPLACEMENT POLYNUCLEOTIDE ASSAY METHOD AND POLYNUCLEOTIDE COMPLEX REAGENT THEREFOR Steven E Diamond, Joseph G Brewen, Jon I Wil- liams, Marian S Ellwood, Mary Collins, Edward F Fritsch assigned to Allied Corporation; Gene- tics Institute Inc into the (P)/(L) binding region. A volume ex- cluding polymeric agent such as poly(ethylene oxide) (PEO or PEG) or other polyethers en- hances the rate of appearance of displaced labeled polynucleotide. Determination of dis- placed labeled polynucleotide (L) gives a value which is a function of the presence and con- centration of target nucleotide sequence (G) in the sample. 4766066 A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) con- tains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the tar- get nucleotide sequence (G) causes binding, ini- tially between G and a single-stranded portion (IBR) of the target binding region (TBR). There- after the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) into the (P)/(L) binding region. Determination of displaced labeled polynucleotide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample. METHOD OF USING BACTERIOPHAGE LAMBDA P1 PROMOTER TO PRODUCE A FUNCTIONAL POLYPEPTIDE IN STREPTOMYCES Stuart A Kuhstoss, R Nagaraja Rao assigned to Eli Lilly and Company The invention relates to a method and cloning vehicle for the expression of a functional poly- peptide in Streptomyces. A recombinant DNA cloning vehicle was genetically engineered to bring the expression of the neomycin phosphotransferase gene under the control of the Escherichia coli bacteriophage lambdapL promoter. 4766064 DISPLACEMENT POLYNUCLEOTIDE ASSAY EMPLOYING POLYETHER AND DIAGNOSTIC KIT Jon I Williams, Marian S Ellwood, Mary Col- lins, Edward F Fritsch, Joseph G Brewen, Steven E Diamond assigned to Allied Corporation; Genetics Institute Inc A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) con- tains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the tar- get nucleotide sequence (G) causes binding, ini- tially between G and a single-stranded portion (IBR) of the target binding region (TBR). There- after the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) 4766067 GENE AMPLIFICATION Debajit K Biswas assigned to President and Fel- lows of Harvard College Hybrid DNA capable of effecting controlled amplification in a host cell of a heterologous gene that determines a specific function. The hybrid DNA has the following three different re- gions of DNA: (1) an amplicon that is inducible by an external stimulus; (2) the heterologous gene (or a site for its insertion); and (3) a selec- table marker enabling isolation of the transformed host cells. In order to make a desired compound coded by the heterologous gene, a vehicle including the hybrid DNA is in- troduced into a host cell is transformed with the vector, transformed host cells are grown to a desired population, the external stimulus is con- trolled to effect amplification of the gene in members of the population, cells containing am- plified gene are cultured, and the compound is recovered.

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Page 1: 4766062 Displacement polynucleotide assay method and polynucleotide complex reagent therefor

PATENT ABSTRACTS 63

4766062

D I S P L A C E M E N T P O L Y N U C L E O T I D E A S S A Y

M E T H O D A N D P O L Y N U C L E O T I D E C O M P L E X

R E A G E N T T H E R E F O R

Steven E Diamond, Joseph G Brewen, Jon I Wil- liams, Marian S Ellwood, Mary Collins, Edward F Fritsch assigned to Allied Corporation; Gene- tics Institute Inc

into the (P)/(L) binding region. A volume ex- cluding polymeric agent such as poly(ethylene oxide) (PEO or PEG) or other polyethers en- hances the rate of appearance of displaced labeled polynucleotide. Determination of dis- placed labeled polynucleotide (L) gives a value which is a function of the presence and con- centration of target nucleotide sequence (G) in the sample.

4766066

A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) con- tains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the tar- get nucleotide sequence (G) causes binding, ini- tially between G and a single-stranded portion (IBR) of the target binding region (TBR). There- after the labeled polynucleotide (L) is displaced from the complex by branch migration of (G) into the (P)/(L) binding region. Determination of displaced labeled polynucleotide (L) gives a value which is a function of the presence and concentration of target nucleotide sequence (G) in the sample.

M E T H O D O F U S I N G B A C T E R I O P H A G E L A M B D A P1

P R O M O T E R T O P R O D U C E A F U N C T I O N A L P O L Y P E P T I D E I N

S T R E P T O M Y C E S

Stuart A Kuhstoss, R Nagaraja Rao assigned to Eli Lilly and Company

The invention relates to a method and cloning vehicle for the expression of a functional poly- peptide in Streptomyces. A recombinant DNA cloning vehicle was genetically engineered to bring the expression of the neomycin phosphotransferase gene under the control of the Escherichia coli bacteriophage lambdapL promoter.

4766064

D I S P L A C E M E N T P O L Y N U C L E O T I D E A S S A Y

E M P L O Y I N G P O L Y E T H E R A N D D I A G N O S T I C K I T

Jon I Williams, Marian S Ellwood, Mary Col- lins, Edward F Fritsch, Joseph G Brewen, Steven E Diamond assigned to Allied Corporation; Genetics Institute Inc

A diagnostic reagent is disclosed containing a complex of a probe polynucleotide (P) bound via purine/pyrimidine hydrogen bonding to a labeled polynucleotide (L). The probe (P) con- tains a target binding region (TBR) capable of binding to a target nucleotide sequence (G) of a biological sample. A method is disclosed in which contact with a sample containing the tar- get nucleotide sequence (G) causes binding, ini- tially between G and a single-stranded portion (IBR) of the target binding region (TBR). There- after the labeled polynucleotide (L) is displaced from the complex by branch migration of (G)

4766067

G E N E A M P L I F I C A T I O N

Debajit K Biswas assigned to President and Fel- lows of Harvard College

Hybrid DNA capable of effecting controlled amplification in a host cell of a heterologous gene that determines a specific function. The hybrid DNA has the following three different re- gions of DNA: (1) an amplicon that is inducible by an external stimulus; (2) the heterologous gene (or a site for its insertion); and (3) a selec- table marker enabling isolation of the transformed host cells. In order to make a desired compound coded by the heterologous gene, a vehicle including the hybrid DNA is in- troduced into a host cell is transformed with the vector, transformed host cells are grown to a desired population, the external stimulus is con- trolled to effect amplification of the gene in members of the population, cells containing am- plified gene are cultured, and the compound is recovered.