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US5405775A Expired / Lapsed US ⏱️ 0.0 years remaining

Retrons coding for hybrid DNA/RNA molecules

Assignee: NEW JERSEY UNIVERSITY HEIGHTS, University of, Medicine and Dentistry of
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Filing: 1990-05-02
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Granted: 1995-04-11
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Expires: 2010-05-02
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Abstract

Abstract A multicopy single-stranded DNA (msDNA) synthesizing system in E. coli is disclosed. The use of the msDNA system to synthesize cDNA in vivo is disclosed. Construction of synthetic msDNA is also disclosed. Also processes for gene amplification and for producing a stable RNA are disclosed.

Inventors

  • 👤 Masayori Inouye
  • 👤 Sumiko Inouye

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-07 19:56
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/10; C12N9/12; C12N9/1241; C12N9/1276; C12N15/00; C12N15/09; C12N15/10; C12P; C12P19/00; C12P19/26; C12P19/28; C12P19/30; C12P19/34

Claims Analysis

Total Claims: 32
Claim 1
Independent Claim 106 words
A plasmid which comprises a retron which encodes a msDNA molecule selected from the group consisting of Ec67, Ec74, ms100 and ms101, which has a DNA and an RNA portion, which retron comprises an msd gene, an msr gene and an open reading frame (ORF), the msd gene coding for the DNA strand of the msDNA molecule and the msr gene coding for the RNA strand of the msDNA molecule, the msr gene overlapping with and in opposite orientation with respect to the msd gene and the ORF coding for a reverse transcriptase (RT), said RT synthesizing cDNA from an RNA transcript into the msDNA molecule.
Claim 2
Dependent Claim 106 words
A plasmid which comprises a retron which encodes a msDNA molecule selected from the group consisting of Ec67, Ec74, ms100 and ms101, which has a DNA and an RNA portion, which retron comprises an msd gene, an msr gene and an open reading frame (ORF), the msd gene coding for the DNA strand of the msDNA molecule and the msr gene coding for the RNA strand of the msDNA molecule, the msr gene overlapping with and in opposite orientation with respect to the msd gene and the ORF coding for a reverse transcriptase (RT), said RT synthesizing cDNA from an RNA transcript into the msDNA molecule.
Claim 3
Dependent Claim 11 words
The plasmid of claim 1 wherein the msDNA molecule is Ec67.
Claim 4
Dependent Claim 11 words
The plasmid of claim 1 wherein the msDNA molecule is Ec74.
Claim 5
Dependent Claim 11 words
The plasmid of claim 1 wherein the msDNA molecule is ms100.
Claim 6
Dependent Claim 11 words
The plasmid of claim 1 wherein the msDNA molecule is ms101.
Claim 7
Dependent Claim 30 words
The plasmid of claim 1 wherein the msDNA molecule contains a foreign DNA fragment positioned in an antisense orientation in the RNA or the DNA portion of the msDNA molecule.
Claim 8
Dependent Claim 11 words
The plasmid of claim 6 wherein the msDNA molecule is ms100.
Claim 9
Dependent Claim 11 words
The plasmid of claim 6 wherein the msDNA molecule is ms101.
Claim 10
Dependent Claim 9 words
A prokaryote transformed with the plasmid of claim 1.
Claim 11
Dependent Claim 9 words
The prokaryote of claim 9 which is a bacterium.
Claim 12
Dependent Claim 9 words
The bacterium of claim 10 which is E. coli.
Claim 13
Dependent Claim 109 words
An isolated retron which encodes a hybrid DNA/RNA msDNA molecule selected from the group consisting of Ec67, Ec74, ms100 and ms101, which has a DNA and an RNA portion, which retron comprises an msd gene, an msr gene and an open reading frame (ORF), the msd gene coding for the DNA strand of the msDNA molecule and the msr gene coding for the RNA strand of the msDNA molecule, the msr gene overlapping with and in opposite orientation with respect to the msd gene and the ORF coding for a reverse transcriptase (RT), said RT synthesizing cDNA from an RNA transcript of the retron which encodes the msDNA molecule.
Claim 14
Dependent Claim 109 words
An isolated retron which encodes a hybrid DNA/RNA msDNA molecule selected from the group consisting of Ec67, Ec74, ms100 and ms101, which has a DNA and an RNA portion, which retron comprises an msd gene, an msr gene and an open reading frame (ORF), the msd gene coding for the DNA strand of the msDNA molecule and the msr gene coding for the RNA strand of the msDNA molecule, the msr gene overlapping with and in opposite orientation with respect to the msd gene and the ORF coding for a reverse transcriptase (RT), said RT synthesizing cDNA from an RNA transcript of the retron which encodes the msDNA molecule.
Claim 15
Dependent Claim 13 words
The retron of claim 12, wherein the msr and msd genes are synthetic.
Claim 16
Dependent Claim 12 words
The retron of claim 13 which codes for the ms100 msDNA molecule.
Claim 17
Dependent Claim 19 words
The retron of claim 12 wherein the ORF is located upstream of the msd and downstream of the msr.
Claim 18
Dependent Claim 19 words
The retron of claim 15, wherein the ORF is from a source different than the msr and msd genes.
Claim 19
Dependent Claim 13 words
The retron of claim 15 wherein the RT has 586 amino acid residues.
Claim 20
Dependent Claim 12 words
The retron of claim 12 which codes for the Ec67 msDNA molecule.
Claim 21
Dependent Claim 27 words
The retron of claim 12, wherein a foreign nucleic acid sequence is positioned in the gene selected from the group consisting of the msr and msd genes.
Claim 22
Dependent Claim 12 words
The retron of claim 12 which codes for the Ec74 msDNA molecule.
Claim 23
Dependent Claim 12 words
The retron of claim 12 which codes for the ms101 msDNA molecule.
Claim 24
Dependent Claim 30 words
The retron of claim 12 wherein the msDNA molecule contains a foreign DNA fragment positioned in an antisense orientation in the RNA or the DNA portion of the msDNA molecule.
Claim 25
Dependent Claim 11 words
The retron of claim 22 wherein the msDNA molecule is ms100.
Claim 26
Dependent Claim 11 words
The retron of claim 22 wherein the msDNA molecule is ms101.
Claim 27
Dependent Claim 15 words
An isolated msDNA molecule selected from the group consisting of Ec67, Ec74, ms100 and ms101.
Claim 28
Dependent Claim 15 words
An isolated msDNA molecule selected from the group consisting of Ec67, Ec74, ms100 and ms101.
Claim 29
Dependent Claim 10 words
The msDNA of claim 25 which is the synthetic ms100.
Claim 30
Dependent Claim 8 words
The msDNA of claim 25 which is Ec67.
Claim 31
Dependent Claim 8 words
The msDNA of claim 25 which is Ec74.
Claim 32
Dependent Claim 10 words
The msDNA of claim 25 which is the synthetic ms101.

References Cited (Backward Citations)

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