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US5405775A
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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
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 20:17
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)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1998-04-10 | 1998-10-10 | 1999-04-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2002-04-10 | 2002-10-10 | 2003-04-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2006-04-10 | 2006-10-10 | 2007-04-11 | $7700.00 | Window Closed / Lapsed |