DNA sequences encoding hybrid lymphoblastoid-leukocyte human interferons
AssigneeUniversity Patents, Inc.
InventorMarvin H. Caruthers
Life of this patent
Estimating the term…
Abstract
DNA sequences encoding for novel human lymphoblastoid-leukocyte hybrid interferons. Methods of making nucleotide sequences comprising ribosome binding sites, promoter regions, and segments of human lymphoblastoid and leukocyte interferons are described. DNA sequences for hybrid interferons are cloned and expressed in plasmid vectors. Transformed microorganisms and novel lymphoblastoid-leukocyte polypeptide interferons having biological or immunological acitivity are disclosed.
Claims31
A deoxynucleotide sequence encoding for a hybrid lymphoblastoid-leukocyte human interferon comprising 166 amino acids and optionally having an additional methionine attached to the ordinary first amino acid at the N-terminus with a plus strand represented by the following formula:
- 5'-ATG-W-X-P-Y-B-Z
- wherein W is TGT or TGC;
- X consists essentially of deoxynucleotides encoding for amino acids 2 to 46 of human lymphoblastoid interferon (LbIF);
- Y consists essentially of deoxynucleotides encoding for amino acids 60 to 85 of said LbIF;
- Z consists essentially of deoxynucleotides encoding for amino acids 112 to 166 of said LbIF;
- P consists essentially of deoxynucleotides encoding for amino acids 47 to 59 of human .alpha.C-, .alpha.F-, or .alpha.I-type leukocyte interferon (LeIF) and is CAG.sub.47 TTC CAA AAG.sub.50 GCT CAG GCA ATC TCG.sub.55 GTT CTG CAT GAG.sub.59, or CAG.sub.47 TTC CAA AAG.sub.50 ACC CAG GCA ATC TCG.sub.55 GTT CTG CAT GAG.sub.59 ;
- B consists essentially of deoxynucleotides encoding for amino acids 86 to 111 to said .alpha.C-, .alpha.F-, or .alpha.I-type LeIF and is TCC.sub.86 ACT GAA CTC TAT.sub.90 CAG CAA CTG AAC GAT.sub.95 CTT GAA GCT TGC GTT.sub.100 ATC CAG GAG GTA GGC.sub.105 GTT GAA GAG ACT CCG.sub.110 CTT, TCC.sub.86 ATC GAA CTC AAT.sub.90 CAG CAA CTG AAC GAT.sub.95 ATG GAA GCT TGC GTT.sub.100 ATC CAG GAG GTA GGC.sub.105 GTT GAA GAG ACT CCG.sub.110 CTT, or TCC.sub.86 ATC GAA CTC TAT.sub.90 CAG CAA CTA AAC AAC.sub.95 CTT GAA GCT TGC GTT.sub.100 ATC CAG GAT GTA GGC.sub.105 ATG GAA GAG ACT CCG.sub.110 CTT;
- and transcriptional and translational single or multiple base substitutions, insertions and inversions;
- wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl and T is thymidyl.
The deoxynucleotide sequence according to claim 2, 5'-ATG TGT.sub.1 GAT TTG CCG CAG.sub.5 ACT CAT AGC TTG GGC.sub.10 AAC CGA AGA GCA CTC.sub.15 ATC CTG TTG GCC CAA.sub.20 ATG GGT CGC ATT TCC.sub.25 CTG TTC TCG TGC CTT.sub.30 AAA GAC CGC CAC GAT.sub.35 TTC GGG TTT CCA CAG.sub.40 GAA GAG TTT GAC GGC.sub.45 AAT CAG TTC CAA AAG.sub.50 GCT CAG GCA ATC TCG.sub.55 GTT CTG CAT GAG ATG.sub.60 ATT CAG CAA ATC TTC.sub.65 AAC CTG TTC TCC ACT.sub.70 AAA GAC TCT TCG GCT.sub.75 GCT TGG AAC GAA TCC.sub.80 TTG CTT GAT AAA TTC.sub.85 TCC ACT GAA CTC TAT.sub.90 CAG CAA CTG AAC GAT.sub.95 CTT GAA GCT TGC GTT.sub.100 ATC CAG GAG GTA GGC.sub.105 GTT GAA GAG ACT CCG.sub.110 CTT ATG AAT GTT GAG.sub.115 TCT ATC CTG GCT GTA.sub.120 TCT AAG TAT TTT CAG.sub.125 CGA ATT ACC CTC TAC.sub.130 CTC AGT GAG AAG AAA.sub.135 TAT TCA CCG TGT GCG.sub.140 TGG GAA ATT GTG AGA.sub.145 GCC GAA ATC ATG CGT.sub.150 TCC CTG ACT CTT CTG.sub.155 ACG AAC CTC CAG GAA.sub.160 CGC CTG CGT AAT AAA.sub.165 GAC.
The deoxynucleotide sequence according to claim 2, 5'-ATG TGT.sub.1 GAT TTG CCG CAG.sub.5 ACT CAT AGC TTG GGC.sub.10 AAC CGA AGA GCA CTC.sub.15 ATC CTG TTG GCC CAA.sub.20 ATG GGT CGC ATT TCC.sub.25 CTG TTC TCG TGC CTT.sub.30 AAA GAC CGC CAC GAT.sub.35 TTC GGG TTT CCA CAG.sub.40 GAA GAG TTT GAC GGC.sub.45 AAT CAG TTC CAA AAG.sub.50 GCT CAG GCA ATC TCG.sub.55 GTT CTG CAT GAG ATG.sub.60 ATT CAG CAA ATC TTC.sub.65 AAC CTG TTC TCC ACT.sub.70 AAA GAC TCT TCG GCT.sub.75 GCT TGG AAC GAA TCC.sub.80 TTG CTT CAT AAA TTC.sub.85 TCC ATC GAA CTC AAT.sub.90 CAG CAA CTG AAC GAT.sub.95 ATG GAA GCT TGC GTT.sub.100 ATC CAG GAG GTA GGC.sub.105 GTT GAA GAG ACT CCG.sub.110 CTT ATG AAT GTT GAG.sub.115 TCT ATC CTG GCT GTA.sub.120 TCT AAG TAT TTT CAG.sub.125 CGA ATT ACC CTC TAC.sub.130 CTC AGT GAG AAG AAA.sub.135 TAT TCA CCG TGT GCG.sub.140 TGG GAA ATT GTG AGA.sub.145 GCC GAA ATC ATG CGT.sub.150 TCC CTG ACT CTT CTG.sub.155 ACG AAC CTC CAG GAA.sub.160 CGC CTG CGT AAT AAA.sub.165 GAC.
The deoxynucleotide sequence according to claim 2, 5'-ATG TGT.sub.1 GAT TTG CCG CAG.sub.5 ACT CAT AGC TTG GGC.sub.10 AAC CGA AGA GCA CTC.sub.15 ATC CTG TTG GCC CAA.sub.20 ATG GGT CGC ATT TCC.sub.25 CTG TTC TCG TGC CTT.sub.30 AAA GAC CGC CAC GAT.sub.35 TTC GGG TTT CCA CAG.sub.40 GAA GAG TTT GAC GGC.sub.45 AAT CAG TTC CAA AAG.sub.50 GCT CAG GCA ATC TCG.sub.55 GTT CTG CAT GAG ATG.sub.60 ATT CAG CAA ATC TTC.sub.65 AAC CTG TTC TCC ACT.sub.70 AAA GAC TCT TCG GCT.sub.75 GCT TGG AAC GAA TCC.sub.80 TTG CTT GAT AAA TTC.sub.85 TCC ATC GAA CTC TAT.sub.90 CAG CAA CTG AAC AAC.sub.95 CTT GAA GCT TGC GTT.sub.100 ATC CAG GAT GTA GGC.sub.105 ATG GAA GAG ACT CCG.sub.110 CTT ATG AAT GTT GAG.sub.115 TCT ATC CTG GCT GTA.sub.120 TCT AAG TAT TTT CAG.sub.125 CGA ATT ACC CTC TAC.sub.130 CTC AGT GAG AAG AAA.sub.135 TAT TCA CCG TGT GCG.sub.140 TGG GAA ATT GTG AGA.sub.145 GCC GAA ATC ATG CGT.sub.150 TCC CTG ACT CTT CTG.sub.155 L ACG AAC CTC CAG GAA.sub.160 CGC CTG CGT AAT AAA.sub.165 GAC.
A deoxynucleotide sequence encoding for a hybrid lymphoblastoid-leukocyte human interferon comprising a hybrid promoter region, an operator region, a ribosome binding site and a polypeptide-coding-region coding for a polypeptide of 166 amino acids and optionally coding for an additional methionine attached to the ordinary first amino acid at the N-terminus of said polypeptide, and a termination region with a plus strand represented by the following formula:
- 5'-L-O-Q-ATG-W-X-P-Y-B-Z-E
- wherein L consists essentially of the promoter region;
- O consists essentially of the operator region;
- Q is a portion of a ribosome binding site and is A-A-A-A-A-T-T-A-A-G-G-A-G-G-A-T-C-A-C-T, T-T-A-T-T-A-C-C-C-A-A-C-T-T-G-A-G-G-A-A-T-T-T-A-T-A, or C-G-G-C-C-C-C-T-T-A-C-T-T-G-A-G-G-A-T-A-A-A-T-T;
- W is TGT or TGC;
- X consists essentially of deoxynucleotides encoding for amino acids 2 to 46 of human lymphoblastoid interferon (LbIF);
- Y consists essentially of deoxynucleotides encoding for amino acids 60 to 85 of said LbIF;
- Z consists essentially of deoxynucleotides encoding for amino acids 112 to 166 of said LbIF;
- P consists essentially of deoxynucleotides encoding for amino acids 47 to 59 of human .alpha.C-, .alpha.F-, or .alpha.I-type leukocyte interferon (LeIF) and is CAG TTC CAA AAG.sub.50 GCT CAG GCA ATC TCG.sub.55 GTT CTG CAT GAG, or CAG TTC CAA AAG.sub.50 ACC CAG GCA ATC TCG.sub.55 GTT CTG CAT GAG;
- B consists essentially of deoxynucleotides encoding for amino acids 86 to 111 of said .alpha.C-, .alpha.F-, or .alpha.I-type LeIF and is TCC ACT GAA CTC TAT.sub.90 CAG CAA CTG AAC GAT.sub.95 CTT GAA GCT TGC GTT.sub.100 ATC CAG GAG GTA GGC.sub.105 GTT GAA GAG ACT CCG.sub.110 CTT, TCC ATC GAA CTC AAT.sub.90 CAG CAA CTG AAC GAT.sup.95 ATG GAA GCT TGC GTT.sub.100 ATC CAG GAG GTA GGC.sub.105 GTT GAA GAG ACT CCG.sub.110 CTT, or TCC ATC GAA CTC TAT.sub.90 CAG CAA CTG AAC AAC.sub.95 CTT GAA GCT TGC GTT.sub.100 ATC CAG GAT GTA GGC.sub.105 ATG GAA GAG ACT CCG.sub.110 CTT;
- E consists essentially of the termination region;
- and transcriptional and translational single or multiple base substitutions, insertions and inversions;
- wherein A is deoxyadenyl, G is deoxyguanyl, C is deoxycytosyl and T is thymidyl.
Dependencies are inferred from the claim text.