Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
96 words
In a method for preparing a polypeptide in a cellular host, where the polypeptide is heterologous to the host and may be produced in low percentage amounts of total protein, the improvement which comprises: joining an open reading frame DNA sequence coding for said polypeptide with a second open reading frame DNA sequence coding for heterologous superoxide dismutase to form a fusion polypeptide; introducing the sequence coding for said fusion polypeptide under conditions for expression in said host, whereby said fusion polypeptide is expressed; and isolating said fusion polypeptide to provide the polypeptide in high yield.
Claim 2
Dependent Claim
96 words
In a method for preparing a polypeptide in a cellular host, where the polypeptide is heterologous to the host and may be produced in low percentage amounts of total protein, the improvement which comprises: joining an open reading frame DNA sequence coding for said polypeptide with a second open reading frame DNA sequence coding for heterologous superoxide dismutase to form a fusion polypeptide; introducing the sequence coding for said fusion polypeptide under conditions for expression in said host, whereby said fusion polypeptide is expressed; and isolating said fusion polypeptide to provide the polypeptide in high yield.
Claim 3
Dependent Claim
15 words
A method according to claim 1, wherein said superoxide dismutase is human superoxide dismutase (hSOD).
Claim 4
Dependent Claim
11 words
A method according to claim 2, wherein said host is eukaryotic.
Claim 5
Dependent Claim
12 words
A method according to claim 3, wherein said eukaryotic host is yeast.
Claim 6
Dependent Claim
13 words
A method according to claim 2 wherein said polypeptide is a mammalian polypeptide.
Claim 7
Dependent Claim
22 words
A method according to claim 5, wherein said mammalian polypeptide is 3' to said hSOD coding sequence in the direction of transcription.
Claim 8
Dependent Claim
24 words
A method according to claim 5, wherein said mammalian polypeptide is 5' to said DNA sequence coding for SOD in the direction of transcription.
Claim 9
Dependent Claim
18 words
A method according to claim 5, wherein said conditions for expression include an inducible transcriptional initiation regulatory region.
Claim 10
Dependent Claim
34 words
A method according to claim 8, where said transcriptional intiation regulator region consists essentially of, reading in a 5' to 3' direction, a yeast glycolytic enzyme promoter region and adjacent alcohol dehydrogenase-2 control region.
Claim 11
Dependent Claim
11 words
A method according to claim 2, wherein said host is prokaryotic.
Claim 12
Dependent Claim
13 words
A method according to claim 10, wherein said prokaryotic host is E. coli.
Claim 13
Dependent Claim
50 words
A DNA sequence coding for superoxide dismutase joined to a DNA sequence coding for a mammalian polypeptide, where the two coding sequences are joined by bases coding for amino acids defining a selectively cleavable link having at least one amino acid, wherein said link provides for a selectively cleavable site.
Claim 14
Dependent Claim
50 words
A DNA sequence coding for superoxide dismutase joined to a DNA sequence coding for a mammalian polypeptide, where the two coding sequences are joined by bases coding for amino acids defining a selectively cleavable link having at least one amino acid, wherein said link provides for a selectively cleavable site.
Claim 15
Dependent Claim
14 words
A DNA sequence according to claim 12, wherein said cleavable link codes for methionine.
Claim 16
Dependent Claim
13 words
A DNA sequence according to claim 12, wherein said cleavable link is Lys-Arg.
Claim 17
Dependent Claim
15 words
A DNA sequence according to claim 12, wherein said cleavable link is (ASp) 4 Lys.
Claim 18
Dependent Claim
15 words
A DN.A sequence according to claim 12, wherein said cleavable link includes hinge amino acids.
Claim 19
Dependent Claim
16 words
A DNA sequence according to claim 12, wherein said cleavable link is an enzymatically removable link.
Claim 20
Dependent Claim
23 words
An expression sequence including, in the direction of transcription, an inducible transcriptional initiation regulatory region and a DNA sequence according to claim 10.
Claim 21
Dependent Claim
34 words
A DNA sequence coding for superoxide dismutase joined to a DNA sequence coding for a mammalian polypeptide, where the two coding sequences are joined by bases coding for amino acids defining a hinge sequence.
Claim 22
Dependent Claim
34 words
A DNA sequence coding for superoxide dismutase joined to a DNA sequence coding for a mammalian polypeptide, where the two coding sequences are joined by bases coding for amino acids defining a hinge sequence.
Claim 23
Dependent Claim
18 words
The DNA sequence according to claim 19 wherein said hinge sequence is from one to fifteen amino acids.
Claim 24
Dependent Claim
30 words
The DNA sequence according to claim 20 wherein said hinge sequence is composed of nonpolar amino acids consisting of the group selected from glycine, alanine, proline, valine, isoleucine and leucine.
Claim 25
Dependent Claim
28 words
The DNA sequence according to claim 20 wherein said hinge sequence is composed of polar amino acids consisting of the group selected from asparagine, glutamine, serine and threonine.