Claims Analysis
Total Claims: 13
Claim 1
Independent Claim
16 words
A bi-functional, hybrid cosmid cloning vector capable of replication in both Escherichia coli and in Streptomycetes.
Claim 2
Dependent Claim
16 words
A bi-functional, hybrid cosmid cloning vector capable of replication in both Escherichia coli and in Streptomycetes.
Claim 3
Dependent Claim
8 words
The cosmid cloning vector identified herein as pPZ74.
Claim 4
Dependent Claim
8 words
The cosmid cloning vector identified herein as pPZ74.
Claim 5
Independent Claim
94 words
A method of constructing a cosmid cloning vector capable of replication in both Escherichia coli and in Streptomycetes, which comprises the steps of: (a) cleaving a broad host-range Streptomycete plasmid vector at a particular endonuclease-sensitive restriction site; (b) ligating DNA from a plasmid from an E. coli host to the cleaved plasmid to form a hybrid vector having two of said sites; and (c) inserting cohesive-end sequences excised from a suitable source into said hybrid vector by cleaving the latter at a suitable site and ligating the cohesive-end sequences to the cleaved hybrid vector.
Claim 6
Independent Claim
94 words
A method of constructing a cosmid cloning vector capable of replication in both Escherichia coli and in Streptomycetes, which comprises the steps of: (a) cleaving a broad host-range Streptomycete plasmid vector at a particular endonuclease-sensitive restriction site; (b) ligating DNA from a plasmid from an E. coli host to the cleaved plasmid to form a hybrid vector having two of said sites; and (c) inserting cohesive-end sequences excised from a suitable source into said hybrid vector by cleaving the latter at a suitable site and ligating the cohesive-end sequences to the cleaved hybrid vector.
Claim 7
Dependent Claim
26 words
A method according to claim 3, in which said Streptomycete plasmid vector contains a single restriction site which will be unique in said cosmid cloning vector.
Claim 8
Dependent Claim
17 words
A method according to claim 4, in which said single restriction site is a Bgl II site.
Claim 9
Dependent Claim
32 words
A method according to claim 5, in which said vector is the plasmid pPZ12 into which a Bgl II site has been inserted in place of the single SstI site therein. ,
Claim 10
Independent Claim
32 words
A method according to any of claims 3 to 6, in which said particular endonuclease-sensitive restriction site at which said Streptomycete plasmid vector is cleaved in step (a) is a PstI-sensitive site.
Claim 11
Independent Claim
32 words
A method according to any of claims 3 to 7, in which said DNA from a plasmid from an E. coli host used in step (b) is DNA from the plasmid pBR325.
Claim 12
Independent Claim
26 words
A method according to any of claims 3 to 8, in which said cohesive end sequences used in step (c) are excised from the plasmid pBTI-l.
Claim 13
Dependent Claim
27 words
A cosmid cloning vector capable of replication in both Escherichia coli and in Streptomycetes, constructed by a method as claimed in any of claims 3 to 9.