EP0068740A2
Expired / Lapsed
EP
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Recombinant DNA cloning vectors and the eukaryotic and prokaryotic transformants thereof
Assignee: Eli Lilly and Co
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Filing: 1982-06-17
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Granted: —
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Expires: 2002-06-17
Abstract
Abstract A recombinant DNA cloning vector comprising: a) a eukaryotic promoter, b) one or two different structural genes and associated control sequence that convey resistance to either or both antibiotics hygromycin 8 and G418 when transformed into a host cell that is sensitive to either or both antibiotics for which resistance is conveyed, said host cell being susceptible to transformation, cell division, and culture, and c) a prokaryotic replicon, said replicon being functional when said host cell is prokaryotic. subject to the limitations that the one or two structural genes and associated control sequence are adjacent to and, in a eukaryotic host cell, transcribed from the eukaryotic promoter, that a single gene and associated control sequence conveys resistance to either but not both hygromycin 8 and G418, and that the gene conveying resistance to G418 does not code for the enzyme phosphotransferase.
Inventors
- 👤 Ramachandra Nagaraja Rao
- 👤 Robert Frank Santerre
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-06 01:00
CPC Classifications
C; C12; C12N; C12N15/00
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
141 words
A recombinant DNA cloning vector comprising: a) a eukaryotic promoter, b) one or two different structural genes and associated control sequence that convey resistance to either or both antibiotics hygromycin B and G418 when transformed into a host cell that is sensitive to either or both antibiotics for which resistance is conveyed, said host cell being susceptible to transformation, cell division, and culture, and c) a prokaryotic replicon, said replicon being functional when said host cell is prokaryotic, subject to the limitations that the one or two structural genes and associated control sequence are adjacent to and, in a eukaryotic host cell, transcribed from the eukaryotic promoter, that a single gene and associated control sequence conveys resistance to either but not both hygromycin B and G418, and that the gene conveying resistance to G418 does not code for the enzyme phosphotransferase.
Claim 2
Dependent Claim
141 words
A recombinant DNA cloning vector comprising: a) a eukaryotic promoter, b) one or two different structural genes and associated control sequence that convey resistance to either or both antibiotics hygromycin B and G418 when transformed into a host cell that is sensitive to either or both antibiotics for which resistance is conveyed, said host cell being susceptible to transformation, cell division, and culture, and c) a prokaryotic replicon, said replicon being functional when said host cell is prokaryotic, subject to the limitations that the one or two structural genes and associated control sequence are adjacent to and, in a eukaryotic host cell, transcribed from the eukaryotic promoter, that a single gene and associated control sequence conveys resistance to either but not both hygromycin B and G418, and that the gene conveying resistance to G418 does not code for the enzyme phosphotransferase.
Claim 3
Dependent Claim
83 words
The vector of claim 1 which is a plasmid wherein the prokaryotic replicon is selected from the group consisting of the pBR322 replicon, pMBl, NR1, RK2, RK6, pSC101, RPl, RP4, RSF1010, PUB110, and SLP1.2 and the eukaryotic promoter is independently selected from the group consisting of the SV40 early promoter, SV40 late promoter, HSVITK promoter, adenovirus promoter, Ad 2 promoter, polyoma promoter, mouse sarcoma virus promoter, yeast trp-1 promoter, yeast leu 2 promoter, yeast his 3 promoter, and the yeast cytochrome c promoter.
Claim 4
Dependent Claim
17 words
The vector of claim 1 or 2 which comprises the 7.5kb BglII restriction fragment of plasmid pKC203.
Claim 5
Dependent Claim
22 words
The vector of claim 3 wherein the prokaryotic replicon is the pBR322 replicon and the eukaryotic promoter is the SV40 early promoter.
Claim 6
Dependent Claim
22 words
The vector of claim 3 wherein the prokaryotic replicon is the pBR322 replicon and the eukaryotic promoter is the Ad 2 promoter.
Claim 7
Dependent Claim
23 words
The vector of claim 3 wherein the prokaryotic replicon is the pBR322 replicon and the eukaryotic promoter is the yeast cytochrome c promoter.
Claim 8
Dependent Claim
37 words
The vector of claim 1 or 2 which is a plasmid and wherein the genes and associated control sequence convey resistance to both hygromycin B and G418 and comprise the 2.75kb BglII/SalI restriction fragment of plasmid pKC203.
Claim 9
Dependent Claim
23 words
The vector of claim 1 or 2 which is a plasmid and wherein the number of said structural genes is limited to one.
Claim 10
Dependent Claim
27 words
The vector of claim 8 wherein the gene and associated control sequence confers resistance to hygromycin B and comprises the 1.5lkb SacI/BglII restriction fragment of plasmid pKC222.
Claim 11
Dependent Claim
26 words
The vector of claim 8 wherein the gene and associated control sequence confers resistance to G418 and comprises the 1.65kb EcoRI/SalI restriction fragment of plasmid pKC222.
Claim 12
Dependent Claim
31 words
The vector of claim 1 or 2 which is a plasmid selected from the group consisting of plasmid pKC214, pKC215, pGDl, pGD2, pGD3, pGD4, pGD10, pGD11, pGD12, pGDl3, pGD14, and pGDl5.
Claim 13
Dependent Claim
29 words
The vector of claim 1 or 2 which is a plasmid selected from the group consisting of plasmid pL0378, pKC273, pL0314, pL0315, pL0316, pL0317, pL0318, pL0319, pL0320, and pL0321.
Claim 14
Dependent Claim
19 words
A transformed host cell which comprises the recombinant DNA cloning vector of any one of claims 1 to 12.
Claim 15
Dependent Claim
12 words
The transformed host cell of claim 13 which is a prokaryotic cell.
Claim 16
Dependent Claim
12 words
The transformed host cell of claim 13 which is a eukaryotic cell.
Claim 17
Dependent Claim
33 words
The transformed host cell of claim 14 which is selected from the group consisting of E. coli, E. coli K12, E. coli Kl2 BE827, Bacillus, Bacillus subtilis, Pseudomonas, Agrobacterium, Streptomyces, Staphylococcus, and Streptococcus.
Claim 18
Dependent Claim
21 words
The transformed host cell of claim 15 which is selected from the group consisting of Neurospora, Cephalosporium, Aspergillus, Penicillium, and yeast.
Claim 19
Dependent Claim
25 words
The transformed host cell of claim 15 which is selected from the group consisting of a plant cell, an amphibian cell and a mammalian cell.
Claim 20
Dependent Claim
42 words
A restriction fragment selected from the group consisting of the 7.5kb BglII restriction fragment of plasmid pKC203, the 2.75kb BglII/Sall restriction fragment of plasmid pKC203, the 1.51kb SacI/BglII restriction fragment of plasmid pKC222, and the 1.65kb EcoRI/Sall restriction fragment of plasmid pKC222.
Claim 21
Dependent Claim
42 words
A restriction fragment selected from the group consisting of the 7.5kb BglII restriction fragment of plasmid pKC203, the 2.75kb BglII/Sall restriction fragment of plasmid pKC203, the 1.51kb SacI/BglII restriction fragment of plasmid pKC222, and the 1.65kb EcoRI/Sall restriction fragment of plasmid pKC222.
Claim 22
Dependent Claim
27 words
A plasmid comprising the restriction fragment of claim 19 which is selected from the group consisting of plasmid pKC203, pKC222, pKC257, pKC259, pKC261, pKC264, pKC275 and pSC701.
Claim 23
Dependent Claim
10 words
A transformed host cell comprising the plasmid of claim 20.
Claim 24
Dependent Claim
33 words
The transformed host cell of claim 21 which is selected from the group consisting of E. coli, E. coli K12, E. coli K12 BE827, Bacillus, Bacillus subtilis, Pseudomonas, Agrobacterium, Streptomyces, Staphylococcus, and Streptococcus.
Claim 25
Dependent Claim
30 words
The use of the recombinant DNA vector of any one of claims 1 to 12 in transforming a eukaryotic cell to be cultured for production of post translationally modified polypeptide.
References Cited (Backward Citations)
No citation records indexed.
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