Patent US4863857A successfully refreshed from Google Patents.
US4863857A
Expired / Lapsed
US
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Polypeptide complementary to peptides or proteins having an amino acid sequence or nucleotide coding sequence at least partially known
Assignee: The University of Texas System
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Filing: 1985-03-01
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Granted: 1989-09-05
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Expires: 2005-03-01
Abstract
Abstract A method for determining the amino acid sequence of a polypeptide complementary to at least a portion of an original peptide or protein. In one aspect the method involves: (a) determining a first nucleotide sequence of a first nucleic acid coding for the biosynthesis of at least a portion of the original peptide or protein; (b) ascertaining a second nucleotide sequence of a second nucleic acid which base-pairs with the first nucleotide sequence of the first nucleic acid, the first and second nucleic acids pairing in antiparallel directions; and (c) determining the amino acid sequence of the complementary polypeptide by the second nucleotide sequence when read in the same reading frame as the first nucleotide sequence. The complementary polypeptide whose amino acid sequence is thus determined may be obtained by diverse means such as, for example, chemical synthesis, derivation from a protein or larger polypeptide containing said amino acid sequence, or, when the second nucleic acid is DNA, inserting the second nucleotide sequence into a plasmid to form a recombinant DNA plasmid vector and transforming a unicellular organism therewith to produce a transformant unicellular organism biosynthesizing said complementary polypeptide. The ascertainment of particular nucleotide sequences may be circumvented, in one aspect, by utilizing the relationships of amino acids having complementary hydropathies for substitutions as generally dictated by base-pairing nucleotide complementarity.
Inventors
- 👤 J. Edwin Blalock
- 👤 Eric M. Smith
- 👤 Kenneth L. Bost
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 09:08
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/44; C07K14/00; C07K14/435; C07K14/665; C07K14/695; C12; C12Q; C12Q1/00; C12Q1/68; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/68; G01N33/6878; Y; Y10; Y10S; Y10S435/00; Y10S435/803; Y10S435/8215; Y10S435/948; Y10S930/00; Y10S930/01; Y10S930/12; Y10S930/13; Y10S930/14; Y10S930/141
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
334 words
A method for determining the amino acid sequence of a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein; substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine; substituting in place of each valine of the ascertained amino acid sequence glutamine or histidine; substituting place of each leucine of the ascertained amino acid sequence, asparagine, aspartic acid or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, lysine; substituting in place of each cystine of the ascertained amino acid sequence, threonine; substituting in place of each methionine of the ascertained amino acid sequence, tyrosine; substituting in place of each alanine of the ascertained amino acid sequence, arginine; substituting in place of each arginine of the ascertained amino acid sequence, alanine or serine; substituting in place of each lysine of the ascertained amino acid sequence, phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, leucine; substituting in place of each aspartic acid of the ascertained amino acid sequence, leucine; substituting in place of each glutamine of the ascertained amino acid sequence, valine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine; substituting in place of each histidine of the ascertained amino acid sequence, valine; substituting in place of each glycine of the ascertained amino acid sequence, proline; substituting in place of each threonine of the ascertained amino acid sequence, tryptophan or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, threonine; substituting in place serine of the ascertained amino acid sequences, arginine or retaining said serine; substituting in place of each tyrosine of the ascertained amio acid sequence, isoluecine or methionine; substituting in place of each proline of the ascertained amino acid sequence, glycine; and determining the amino acid sequence obtained after the above substitutions.
Claim 2
Independent Claim
334 words
A method for determining the amino acid sequence of a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein; substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine; substituting in place of each valine of the ascertained amino acid sequence glutamine or histidine; substituting place of each leucine of the ascertained amino acid sequence, asparagine, aspartic acid or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, lysine; substituting in place of each cystine of the ascertained amino acid sequence, threonine; substituting in place of each methionine of the ascertained amino acid sequence, tyrosine; substituting in place of each alanine of the ascertained amino acid sequence, arginine; substituting in place of each arginine of the ascertained amino acid sequence, alanine or serine; substituting in place of each lysine of the ascertained amino acid sequence, phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, leucine; substituting in place of each aspartic acid of the ascertained amino acid sequence, leucine; substituting in place of each glutamine of the ascertained amino acid sequence, valine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine; substituting in place of each histidine of the ascertained amino acid sequence, valine; substituting in place of each glycine of the ascertained amino acid sequence, proline; substituting in place of each threonine of the ascertained amino acid sequence, tryptophan or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, threonine; substituting in place serine of the ascertained amino acid sequences, arginine or retaining said serine; substituting in place of each tyrosine of the ascertained amio acid sequence, isoluecine or methionine; substituting in place of each proline of the ascertained amino acid sequence, glycine; and determining the amino acid sequence obtained after the above substitutions.
Claim 3
Dependent Claim
51 words
The method of claim 1 defined further wherein: serine is substituted in place of each arginine of the ascertained amino acid sequence; serine is retained in place of each serine of the ascertained amino acid sequence; and cysteine is substituted in place of each threonine of the ascertained amino acid sequence.
Claim 4
Dependent Claim
37 words
The method of claim 2 wherein the complementary polypeptide is defined further as retaining complemetarity or binding affinity for to the original peptite or protein regardless of the amino terminal and carboxy-terminal directionality of said complementary polypeptide.
Claim 5
Independent Claim
334 words
A method for obtaining a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine; substituting in place of each valine of the ascertained amino acid sequence glutamine or histidine; substituting place of each leucine of the ascertained amino acid sequence, asparagine, aspartic acid or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, lysine; substituting in place of each cysteine of the ascertained amino acid sequence, threonine; substituting in place of each methionine of the ascertained amino acid sequence, tyrosine; substituting in place of each alanine of the ascertained amino said sequence, arginine; substituting in place of each arginine of the ascertained amino acid sequence, alanine or serine; substituting in place of each lysine of the ascertained amino acid sequence, phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, leucine; substituting in place of each aspartic acid of the ascertained amino acid sequence, leucine; substituting in place of each glutamine of the ascertained amino acid sequence valine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine; substituting in place of each histidine of the ascertained amino acid sequence, valine; substituting in place of each glycine of the ascertained amino acid sequence, proline; substituting in place of each threonine of the ascertained amino acid sequence, tryptophan or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, threonine; substituting in place of each serine of the ascertained amino acid sequence, arginine or retaining said serine; substituting in place of each tyrosine of the ascertained amino acid sequence, isoleucine or methionine; substituting in place of each proline of the ascertained amino acid sequence, glycine; and obtaining a polypeptide comprising the amino acid sequence determined by the above substitutions.
Claim 6
Independent Claim
334 words
A method for obtaining a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine; substituting in place of each valine of the ascertained amino acid sequence glutamine or histidine; substituting place of each leucine of the ascertained amino acid sequence, asparagine, aspartic acid or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, lysine; substituting in place of each cysteine of the ascertained amino acid sequence, threonine; substituting in place of each methionine of the ascertained amino acid sequence, tyrosine; substituting in place of each alanine of the ascertained amino said sequence, arginine; substituting in place of each arginine of the ascertained amino acid sequence, alanine or serine; substituting in place of each lysine of the ascertained amino acid sequence, phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, leucine; substituting in place of each aspartic acid of the ascertained amino acid sequence, leucine; substituting in place of each glutamine of the ascertained amino acid sequence valine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine; substituting in place of each histidine of the ascertained amino acid sequence, valine; substituting in place of each glycine of the ascertained amino acid sequence, proline; substituting in place of each threonine of the ascertained amino acid sequence, tryptophan or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, threonine; substituting in place of each serine of the ascertained amino acid sequence, arginine or retaining said serine; substituting in place of each tyrosine of the ascertained amino acid sequence, isoleucine or methionine; substituting in place of each proline of the ascertained amino acid sequence, glycine; and obtaining a polypeptide comprising the amino acid sequence determined by the above substitutions.
Claim 7
Dependent Claim
51 words
The method of claim 4 defined further wherein: serine is substituted in place of each arginine of the ascertained amino acid sequence; serine is retained in place of each serine of the ascertained amino acid sequence; and cysteine is substituted in place of each threonine of the ascertained amino acid sequence.
Claim 8
Dependent Claim
15 words
The method of claim 4 wherein the obtaining step is comprises chemically synthesizing said polypeptide.
Claim 9
Dependent Claim
24 words
The method of claim 4 wherein the obtaining step comprises excising said polypeptide from a protein or larger polypeptide including said amino acid sequence.
Claim 10
Dependent Claim
48 words
The method of claim 4 wherein the obtaining step is defined further as comprising insertion of a DNA nucleotide sequence coding for said polypeptide into a plasmid to form a recombinant DNA vector and transforming a unicellular organism therewith to produce a transformant unicellular organism biosynthesizing said polypeptide.
Claim 11
Dependent Claim
23 words
The method of claim 8 wherein the unicellular organism is selected from the group consisting of bacterial cells, yeast cells and mammalian cells.
Claim 12
Independent Claim
362 words
A method for determining the amino acid sequence of a polypeptide having complementary binding affinity for to at least a portion of an original protein or peptide, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original protein or peptide; reading the ascertained amino acid sequence, starting from the carboxy-terminal direction thereof, to substitutingly correspond to the amino-terminal direction of the complementary polypeptide; substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine, asparagine or aspartic acid; substituting in place of each leucine of the ascertained amino acid sequence, lysine, glutamine or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, threonine or alanine; substituting in place of each methionine of the ascertained amino acid sequence, histidine; substituting in place of each alanine of the ascertained amino acid sequence, arginine, serine, glycine or cystein; substituting in place of each arginine of the ascertained amino acid sequence, alanine, serine, threonine or proline; substituting in place of each lysine of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, isoleucine or vlaine; substituting in place of each aspartic acid of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each glutamine of the ascertained amino acid sequence, leucine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each histidine of the ascertained amino acid sequence, valine or methionine; substituting in place of each glycine of the ascertained amino acid sequence, proline, serine, threonine or alanine; substituting in place of each threonine of the ascertained amino acid sequence, glycine, serine, arginine or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, proline; substituting in place of each serine of the ascertained amino acid sequence, glycine, threonine, alanine, or arginine; substituting in place of each tyrosine of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each proline of the ascertained amino acid sequence, glycine, arginine or tryptophan; and determining the amino acid sequence found after the above substitutions.
Claim 13
Independent Claim
362 words
A method for determining the amino acid sequence of a polypeptide having complementary binding affinity for to at least a portion of an original protein or peptide, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original protein or peptide; reading the ascertained amino acid sequence, starting from the carboxy-terminal direction thereof, to substitutingly correspond to the amino-terminal direction of the complementary polypeptide; substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine, asparagine or aspartic acid; substituting in place of each leucine of the ascertained amino acid sequence, lysine, glutamine or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, threonine or alanine; substituting in place of each methionine of the ascertained amino acid sequence, histidine; substituting in place of each alanine of the ascertained amino acid sequence, arginine, serine, glycine or cystein; substituting in place of each arginine of the ascertained amino acid sequence, alanine, serine, threonine or proline; substituting in place of each lysine of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, isoleucine or vlaine; substituting in place of each aspartic acid of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each glutamine of the ascertained amino acid sequence, leucine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each histidine of the ascertained amino acid sequence, valine or methionine; substituting in place of each glycine of the ascertained amino acid sequence, proline, serine, threonine or alanine; substituting in place of each threonine of the ascertained amino acid sequence, glycine, serine, arginine or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, proline; substituting in place of each serine of the ascertained amino acid sequence, glycine, threonine, alanine, or arginine; substituting in place of each tyrosine of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each proline of the ascertained amino acid sequence, glycine, arginine or tryptophan; and determining the amino acid sequence found after the above substitutions.
Claim 14
Dependent Claim
180 words
The method of claim 10 defined further wherein: aspartic acid or histidine is substituted in place of each valine of the ascertained amino acid sequence; lysine of glutamine is substituted in place of each leucine of the ascertained amino acid sequence; glutamic acid is substituted in place of each phenylalanine of the ascertained amino acid sequence; alanine or proline is substituted in place of each arginine of the ascertained amino acid sequence; leucine is substituted in place of each lysine of the ascertained amino acid sequence; valine is substituted in place of each histidine of the ascertained amino acid sequence; proline or alanine is substituted in place of each glycine of the ascertained amino acid sequence; glycine or arginine is substituted in place of each threonine of the ascertained amino acid sequence; glycine, arginine or alanine is substituted in place of each serine of the ascertained amino acid sequence; valine is substituted in place of each tyrosine of the ascertained amino acid sequence; and glycine or arginine is substituted in place of each proline of the ascertained amino acid sequence.
Claim 15
Independent Claim
359 words
A method for obtaining a polypeptide having complementary binding affinity for at least a portion of an original peptide or protein, comprising the steps of ascertaining the amino acid sequence of at least a portion of the original protein or peptide; reading the ascertained amino acid sequence starting from the carboxy-terminal direction thereof to substitutingly correspond to the amino-terminal direction of the complementary polypeptide; substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine, asparagine or aspartic acid; substituting in place of each leucine of the ascertained amino acid sequence, lysine, glutamine or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, threonine or alanine; substituting in place of each methionine of the ascertained amino acid sequence, histidine; substituting in place of each alanine of the ascertained amino acid sequence, arginine, serine, glycine or cysteine; substituting in place of each arginine of the ascertained amino acid sequence, alanine, serine, threonine or proline; substituting in place of each lysine of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each aspartic acid of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each glutamine of the ascertained amino acid sequence, leucine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each histidine of the ascertained amino acid sequence, valine or methionine; substituting in place of each glycine of the ascertained amino acid sequence, proline, serine, threonine or alanine; substituting in place of each threonine of the ascertained amino acid sequence, glycine, serine, arginine or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, proline; substituting in place of each serine of the ascertained amino acid sequence, glycine, threonine, alanine or arginine; substituting in place of each tyrosine of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each proline of the ascertained amino acid sequence, glycine, arginine or tryptophan; and obtaining a polypeptide comprising the amino acid sequence determined by the above substitutions.
Claim 16
Independent Claim
359 words
A method for obtaining a polypeptide having complementary binding affinity for at least a portion of an original peptide or protein, comprising the steps of ascertaining the amino acid sequence of at least a portion of the original protein or peptide; reading the ascertained amino acid sequence starting from the carboxy-terminal direction thereof to substitutingly correspond to the amino-terminal direction of the complementary polypeptide; substituting in place of each isoleucine of the ascertained amino acid sequence, tyrosine, asparagine or aspartic acid; substituting in place of each leucine of the ascertained amino acid sequence, lysine, glutamine or glutamic acid; substituting in place of each phenylalanine of the ascertained amino acid sequence, threonine or alanine; substituting in place of each methionine of the ascertained amino acid sequence, histidine; substituting in place of each alanine of the ascertained amino acid sequence, arginine, serine, glycine or cysteine; substituting in place of each arginine of the ascertained amino acid sequence, alanine, serine, threonine or proline; substituting in place of each lysine of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each asparagine of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each aspartic acid of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each glutamine of the ascertained amino acid sequence, leucine; substituting in place of each glutamic acid of the ascertained amino acid sequence, leucine or phenylalanine; substituting in place of each histidine of the ascertained amino acid sequence, valine or methionine; substituting in place of each glycine of the ascertained amino acid sequence, proline, serine, threonine or alanine; substituting in place of each threonine of the ascertained amino acid sequence, glycine, serine, arginine or cysteine; substituting in place of each tryptophan of the ascertained amino acid sequence, proline; substituting in place of each serine of the ascertained amino acid sequence, glycine, threonine, alanine or arginine; substituting in place of each tyrosine of the ascertained amino acid sequence, isoleucine or valine; substituting in place of each proline of the ascertained amino acid sequence, glycine, arginine or tryptophan; and obtaining a polypeptide comprising the amino acid sequence determined by the above substitutions.
Claim 17
Dependent Claim
14 words
The method of claim 12 wherein the obtaining step comprises chemically synthesizing said polypeptide.
Claim 18
Dependent Claim
27 words
The method of claim 12 wherein the obtaining step comprises further as comprising excising said polypeptide from a protein or larger polypeptide including said amino acid sequence.
Claim 19
Dependent Claim
44 words
The method of claim 12 wherein the obtaining step comprises insertion of a DNA nucleotide sequence coding for said polypeptide into a plasmid to form a recombinant DNA vector and transforming a unicellular organism therewith to produce a transformant unicellular organism biosynthesizing said polypeptide.
Claim 20
Dependent Claim
23 words
The method of claim 12 wherein the unicellular organism is selected from the group consisting of bacterial cells, yeast cells and mammalian cells.
Claim 21
Dependent Claim
169 words
The method of claim 12 defined further wherein: substituting in place of each valine of the ascertained amino acid sequence, aspartic acid or histidine; substituting in place of each leucine of the ascertained amino acid sequence, lysine or glutamine; substituting in place of each phenylalanine of the ascertained amino acid sequence, glutamic acid; substituting in place of each orginine of the ascertained amino acid sequence, alanine or proline; substituting in place of each lysine of the ascertained amino acid sequence, leucine; substituting in place of each histidine of the ascertained amino acid sequence, valine; substituting in place of each glycine of the ascertained amino acid sequence, proline or alanine; substituting in place of each threonine of the ascertained amino acid sequence, glycine or arginine; substituting in place of each serine of the ascertained amino acid sequence, glycine, arginine or alanine; substituting in place of each tyrosine of the ascertained amino acid sequence, valine; and substituting in place of each proline of the ascertained amino acid sequence, glycine or arginine.
Claim 22
Independent Claim
149 words
A method for determining the amino acid sequence of a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein wherein the amino acids of the polypeptide, original peptide and original protein are defined as being in groups according to the second base of their codons, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein; substituting in place in place of each A group amino acid, an amino acid of the U group substituting in place of U group amino acid, an amino acid of the A group substituting in place of each C group amino acid, an amino acid of the G group; substituting in place of each G group amino acid, an amino acid of the C group; and determining, after the above substitutions, the resulting amino acid sequence.
Claim 23
Independent Claim
149 words
A method for determining the amino acid sequence of a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein wherein the amino acids of the polypeptide, original peptide and original protein are defined as being in groups according to the second base of their codons, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein; substituting in place in place of each A group amino acid, an amino acid of the U group substituting in place of U group amino acid, an amino acid of the A group substituting in place of each C group amino acid, an amino acid of the G group; substituting in place of each G group amino acid, an amino acid of the C group; and determining, after the above substitutions, the resulting amino acid sequence.
Claim 24
Independent Claim
151 words
A method for obtaining a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein, wherein the amino acids of the polypeptide, original peptide or original protein are defined as contained in groups (U, A, C or G) according to the second base of their codons, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein; substituting in place of each A group amino acid, an amino acid of the U group substituting in place of U group amino acid, an amino acid of the A group substituting in place of each C group amino acid, an amino acid of the G group; and substituting in place of each G group amino acid, an amino acid of the C group; and obtaining a polypeptide comprising the amino acid sequence defined by the above substitutions.
Claim 25
Independent Claim
151 words
A method for obtaining a polypeptide having complementary binding affinity for to at least a portion of an original peptide or protein, wherein the amino acids of the polypeptide, original peptide or original protein are defined as contained in groups (U, A, C or G) according to the second base of their codons, comprising the steps of: ascertaining the amino acid sequence of at least a portion of the original peptide or protein; substituting in place of each A group amino acid, an amino acid of the U group substituting in place of U group amino acid, an amino acid of the A group substituting in place of each C group amino acid, an amino acid of the G group; and substituting in place of each G group amino acid, an amino acid of the C group; and obtaining a polypeptide comprising the amino acid sequence defined by the above substitutions.
Claim 26
Dependent Claim
14 words
The method of claim 19 wherein the obtaining step comprises chemically synthesizing said polypeptide.
Claim 27
Dependent Claim
24 words
The method of claim 19 wherein the obtaining step comprises excising said polypeptide from a protein or larger polypeptide including said amino acid sequence.
Claim 28
Dependent Claim
39 words
The method of claim 19 wherein the obtaining step comprises insertion of a DNA nucleotide sequence including the code for said polypeptide into a plasmid to form a recombinant DNA vector and transforming a unicellular organism biosynthesizing said polypeptide.
Claim 29
Dependent Claim
23 words
The method of claim 22 wherein the unicellular organism is selected from the group consisting of bacterial cells, yeast cells and mammalian cells.
References Cited (Backward Citations)
WO1983004053A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1992-09-04 | 1993-03-06 | 1993-09-05 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1996-09-04 | 1997-03-06 | 1997-09-05 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2000-09-04 | 2001-03-06 | 2001-09-05 | $7700.00 | Window Closed / Lapsed |