EP0048081A2
Expired / Lapsed
EP
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DNA capable of replication and stable mitotic maintenance in a host eukaryote, method of preparation thereof and eukaryotic cell containing same
Assignee: University of California Berkeley
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Filing: 1981-07-10
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Granted: —
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Expires: 2001-07-10
Abstract
Abstract Functioning eukaryotic minichromosomes containing a centromere, at least one replicating site, and at least one functioning structural gene. Centromeres are obtained from a host chromosome and used as a genetic unit in combination with an autonomously replicating segment for high frequency stable transformation of host and other eukaryotic cells. The transformed eukaryotic cells can be cultivated through repeated passages without significant loss of the minichromosome and continued expression of the structural genes present in the minichromosome.
Inventors
- 👤 John Anthony Carbon
- 👤 Louise Broughton Clarke
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-01 18:20
CPC Classifications
C; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C12N15/68
Claims Analysis
Total Claims: 15
Claim 1
Independent Claim
46 words
DNA other than a naturally occurring chromosome capable of replication and stable mitotic maintenance in a host eukaryote comprising a DNA segment coding for chromosome-like activity during mitosis of said host eukaryote and a DNA sequence coding for a replication site recognized by said host eukaryote.
Claim 2
Dependent Claim
46 words
DNA other than a naturally occurring chromosome capable of replication and stable mitotic maintenance in a host eukaryote comprising a DNA segment coding for chromosome-like activity during mitosis of said host eukaryote and a DNA sequence coding for a replication site recognized by said host eukaryote.
Claim 3
Dependent Claim
17 words
DNA as claimed in Claim 1, in which the DNA is also capable of stable meiotic maintenance.
Claim 4
Dependent Claim
24 words
DNA as.claimed in Claim 1, including at least one structural gene capable of expression in said host, said gene being exogenous to said host.
Claim 5
Dependent Claim
17 words
DNA as claimed in any one of Claims 1 to 3, in which the host is yeast.
Claim 6
Dependent Claim
26 words
DNA as claimed in Claim 1, in which the DNA segment coding for chromosome-like activity is derived from a DNA fragment of less than about 2kbp.
Claim 7
Dependent Claim
12 words
A eukaryotic cell having mitotically stable DNA as claimed in Claim 1.
Claim 8
Dependent Claim
31 words
A eukaryotic cell as claimed in Claim 6, in which the DNA includes at least one structural gene capable of expression in the host, the gene being exogenous to the host.
Claim 9
Dependent Claim
54 words
A method for producing a product foreign to a host eukaryotic cell which comprises: transforming the host eukaryote with DNA as claimed in Claim 3 to produce transformants; cultivating the transformants under conditions where the exogenous gene is expressed; and isolating product produced as a result of the expression of the exogenous gene. -
Claim 10
Dependent Claim
58 words
Yeast cells having DNA capable of replication and stable mitotic maintenance comprising a DNA segment coding for chromosome-like activity during mitosis of the yeast and a DNA sequence coding for a replication site recognized by the yeast, in which the DNA is as a result of replication of the in vitro joining of the sequence and the segment.
Claim 11
Dependent Claim
58 words
Yeast cells having DNA capable of replication and stable mitotic maintenance comprising a DNA segment coding for chromosome-like activity during mitosis of the yeast and a DNA sequence coding for a replication site recognized by the yeast, in which the DNA is as a result of replication of the in vitro joining of the sequence and the segment.
Claim 12
Independent Claim
77 words
A method for preparing DNA capable of replication and stable mitotic maintenance in a host eukaryote which comprises: segmenting a chromosome to provide a DNA fragment comprising a centromere and at least one structural gene; cloning the fragment by inserting the fragment into a vector to produce hybrid extrachromosomal DNA and transforming and cultivating a unicellular host for the plasmid; isolating the centromere containing DNA fragment from the plasmid and joining the fragment to an ars gene.
Claim 13
Independent Claim
77 words
A method for preparing DNA capable of replication and stable mitotic maintenance in a host eukaryote which comprises: segmenting a chromosome to provide a DNA fragment comprising a centromere and at least one structural gene; cloning the fragment by inserting the fragment into a vector to produce hybrid extrachromosomal DNA and transforming and cultivating a unicellular host for the plasmid; isolating the centromere containing DNA fragment from the plasmid and joining the fragment to an ars gene.
Claim 14
Dependent Claim
31 words
A DNA fragment derived from a eukaryotic chromosome having chromosome-like activity and imparting mitotic stability when joined to an ars gene and the resulting DNA molecule introduced into a eukaryotic host.
Claim 15
Dependent Claim
31 words
A DNA fragment derived from a eukaryotic chromosome having chromosome-like activity and imparting mitotic stability when joined to an ars gene and the resulting DNA molecule introduced into a eukaryotic host.
References Cited (Backward Citations)
No citation records indexed.
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