SI21397B - Human granulocyte colony stimulating factor. - Google Patents

Abstract

The invention concerned provides a gene which is coding a polypeptide with an activity stimulating factor for a human granulocyte colony (human G-CSF), a recombinant vector of the said built-in gene, a transformed compound which the said vector contains, a polypeptide or glycoprotein having a human G-CSF activity and is manufactured by means of the said transformed compound, and the procedure for recovery of polypeptide or glycoprotein with the human G-CSF activity. The gene from the invention concerned comprises two lines of cDNA, complementary to a messenger RNA (mRNA) coding the polypeptide with the human G-CSF activity and contains also a human chromosome gene coding a polypeptide with the human G-CSF activity

Description

(30) Priority right:

09/17/1985 JP 206066/1985; 09/20/1985 JP 209638/1985; 09/30/1985 JP 217150/1985; 12/02/1985 JP 269455/1985; 02/12/1985 JP 269456/1985; 12/03/1985 JP 270838/1985; 12/03/1985 JP 270839/1985; 07/17/1986 JP 166709/1986; 07/17/1986 JP 166710/1986 (72) Inventors:

(73) Holder:

(74) Representative:

Yamazaki Tatsumi, Setagaya-ku, Tokyo, JP;

Nagata Shigekazu, Ota-ku, Tokyo, JP;

Tsuchiya Masayuki, Toshima-ku, Tokyo, JP;

Hirata Yuichi, Nerima-ku, Tokyo, JP;

Yamamoto Osami, Tokorozawa-shi, Saitama-ken, J P;

Sekimori Yasuo, Kawasaki-shi, Kenagawa-ken, JP

CHUGAI SEIYAKU KABUSHIKI KAISHA,

No. 5-1,5-chome, Ukima, Kita-ku, Tokyo, JP

MARK-INVENTA doo, Rimska 6, p.3916, 1001 Ljubljana, Sl (54) A FACTOR STIMULATING THE HUMAN COLONY OF GRANULOCYTES

Claims (18)

A human chromosomal gene, which is a coding gene for a polypeptide having a factor that stimulates the activity of a human granulocyte colony, characterized in that it contains the nucleotide sequence shown in Figure 5 in whole or in part. The human chromosomal gene according to claim 1, characterized in that said chromosomal gene contains a nucleotide sequence involved in transcriptional control. Human chromosomal gene according to claim 1, characterized in that it is linked to a replicon derived from a microorganism or virus. A recombinant vector comprising the human chromosomal gene of claim 1. A recombinant vector according to claim 4, characterized in that said human chromosomal gene contains a nucleotide sequence that is involved in transcriptional control. A recombinant vector according to claim 4 or 5, characterized in that said human chromosomal gene is linked to a replicon derived from a microorganism or virus. Transformant comprising the recombinant vector of any one of claims 4 to 6. z A process for making a glycoprotein having a factor that stimulates the activity of a human granulocyte colony, characterized in that it comprises the following steps: a) growing the transformant according to claim 7 in a culture medium; and b) obtaining a glycoprotein having a factor that stimulates the activity of a human granulocyte colony. A method according to claim 8, characterized in that said glycoprotein contains a sugar chain moiety and a polypeptide represented by all or part of the following amino acid sequence: Thr Pro Leu Gly Pro Ala Sir Sir Leu Pro Gin Sir Phe Leu Leu Lys Cys Leu Glu Gin Val Arg Lys Ile Gin Gly Asp Gly Ala Ala Leu Gin Glu Lys Leu (Val Sir Glu) _m Cys Ala Thr Tyr Lys Leu Cys His Pro Glu Glu Leu Val Leu Leu Gly His Sir Leu Gly Ile Pro Trp Ala Pro Leu Sir Sir Cys Pro Sir Gin Ala Leu Gin Leu Ala Gly Cys Leu Sir Gin Leu His Sir Gly Leu Phe Leu Tyr Gin Gly Leu Leu Gin Ala Leu Glu Gly Ile Sir Pro Glu Leu Gly Pro Thr Leu Asp Thr Leu Gin Leu Asp Val Ala Asp Phe Ala Thr Thr Ile Trp Gin Gin Met Glu Glu Leu Gly Met Ala Pro Ala Leu Gin Pro Thr Gin Gly Ala Met Pro Ala Phe Ala Sir Ala Phe Gin Arg Arg Ala Gly Gly Val Leu Val Ala Sir His Leu Gin Sir Phe Leu Glu Val Sir Tyr Arg Val Leu Arg His Leu Ala Gin Pro where m is 0 or 1 10. E. coli of type X 1776, characterized in that it harbors a 4-kb DNA fragment encoding for human G-CSF located at the EcoRI site of plasmid pBR327 (FERM BP-956). 11. A cDNA sequence characterized in that it encodes the following amino acid sequence: Met Ala Gly Pro Ala Thr Gin Sir Pro Met Lys Leu Met Ala Leu Gin Leu Leu Leu Trp His Sir Ala Leu Trp Thr Val Gin Glu Ala Thr Pro Leu Gly Pro Ala Sir Sir Leu Pro Gin Sir Phe Leu Leu Lys Cys Leu Glu Gin Val Arg Lys Ile Gin Gly Asp Gly Ala Ala Leu Gin Glu Lys Leu (Val Sir Glu) m Cys Ala Thr Tyr Lys Leu Cys His Pro Glu Glu Leu Val Leu Leu Gly His Sir Leu Gly Ile Pro Trp Ala Pro Leu Sir Sir Cys Pro Sir Gin Ala Leu Gin Leu Ala Gly Cys Leu Sir Gin Leu His Sir Gly Leu Phe Leu Tyr Gin Gly Leu Leu Gin Ala Leu Glu Gly Ile Sir Pro Glu Leu Ala Asp Phe Ala Thr Thr Ile Trp Gin Gin Met Glu Glu Leu Gly Met Ala Pro Ala Leu Gin Pro Thr Gin Gly Ala Met Pro Ala Phe Ala Sir Ala Phe Gin Arg Arg Ala Gly Gly Val Leu Val Ala Sir His Leu Gin Sir Phe Leu Glu W1 Sir Tyr Arg Val Leu Arg His Leu Ala Gin Pro where m is 0 or 1. A cDNA sequence according to claim 11, characterized in that it has the following nucleotide sequence: AT G GCT G GA CCT GCC ACC CAG AGC CCC ATG AAG CTG ATG GCC CGT CAG CTG CTG CTG TGG CAC AGT GCA CTC TGG ACA GTG CAG GAA GCC ACC CCC CTG GGC CCT GCC AGC TCC CTG CCC CAG AGC TTC CTG CTC AAG TGC TTA GAG CAA GTG AGG AAG ATC CAG GGC GAT GGC GCA GCG CTC CAG GAG AAG CTG (GTG AGT GAG) _m TGT GCC ACC TAC AAG CTG TGC CAC CCC GAG GAG CTG GTG CTG CTC GGA CAC TCT CTG GGC AT C CCC TGG GCT CCC CTG AGC AGC TGC CCC AGC CAG GCC CTG CAG CTG GCA GGC TGC TTG AGC CAA CTC CAT AGC GGC CTT TTC CTC TAC CAG GGG CTC CTG CAG GCC CTG GAA GGG ATC TCC CCC GAG TTG GGT CCC ACC TTG GAC ACA CTG CAG CTG GAC CTC GCC GAC TTT GCC ACC ACC ATC TGG CAG CAG ATG GAA GAA CTG GGA ATG GCC CCT GCC CTG CAG CCC ACC CAG GGT GCC ATG CCG GCC TTC GCC TCT GCT TTC CAG CGC CGG GCA GGA GGG GTC CTG GTT GCC TCC CAT CTG CAG AGC TTC CTG GAG GTG TCG TAC CGC GTT CTA CGC CAC CTT GCC CAG CCC where m is 0 or 1. A recombinant vector characterized in that it absorbs the DNA sequence of claim 11 or 12. 14. The transformant comprising the recombinant vector of claim 13. r A method for producing a protein, characterized in that it comprises culturing the transiormant according to claim 14 under suitable conditions and producing said protein. 16. Protein, characterized in that it has the following amino acid sequence: Met Ala Gly Pro Ala Thr Gin Sir Pro Met Lys Leu Met Ala Leu Gin Leu Leu Leu Trp His Sir Ala Leu Trp Thr Val Gin Glu Ala Thr Pro Leu Gly Pro Ala Sir Sir Leu Pro Gin Sir Phe Leu Leu Lys Cys Leu Glu Gin Val Arg Lys Ile Gin Gly Asp Gly Ala Ala Leu Gin Glu Lys Leu (Val Sir Glu) m Cys Ala Thr Tyr Lys Leu Cys His Pro Glu Glu Leu Val Leu Leu Gly His Sir Leu Gly Ile Pro Trp Ala Pro Leu Sir Sir Cys Pro Sir Gin Ala Leu Gin Leu Ala Gly Cys Leu Sir Gin Leu His Sir Gly Leu Phe Leu Tyr Gin Gly Leu Leu Gin Ala Leu Glu Gly Ile Sir Pro Glu Leu Ala Asp Phe Ala Thr Thr Ile Trp Gin Gin Met Glu Glu Leu Gly Met Ala Pro Ala Leu Gin Pro Thr Gin Gly Ala Met Pro Ala Phe Ala Sir Ala Phe Gin Arg Arg Ala Gly Gly Val Leu Val Ala Sir His Leu Gin Sir Phe Leu Glu W4L Sir Tyr Arg Val Leu Arg His Leu Ala Gin Pro where m is 0 or 1. 17. Human granulocyte colony stimulating factor, characterized in that it has the following amino acid sequence: + 1 Thr Pro Leu Gly Pro Ala Sir Sir Leu Pro Gin Sir Phe Leu Leu Lys Cys Leu Glu Gin Val Arg Lys Ile Gin Gly Asp Gly Ala Ala Leu Gin Glu Lys Leu (Val Sir Glu) _m Cys Ala Thr Tyr Lys Leu Cys His Pro Glu Glu Leu Val Leu Leu Gly His Sir Leu Gly Ile Pro Trp Ala Pro Leu Sir Sir Cys Pro Sir Gin Ala Leu Gin Leu Ala Gly Cys Leu Sir Gin Leu His Sir Gly Leu Phe Leu Tyr Gin Gly Leu Leu Gin Ala Leu Glu Gly Ile Sir Pro Glu Leu Gly Pro Thr Leu Asp Thr Leu Gin Leu Asp Val Ala Asp Phe Ala Thr Thr Ile Trp Gin Gin Met Glu Glu Leu ciy Met Ala Pro Ala Leu Gin Pro Thr Gin Gly Ala Met Pro Ala Phe Ala Sir Ala Phe Gin Arg Arg Ala Gly Gly Val Leu Val Ala Sir His Leu Gin Sir Phe Leu Glu Val Sir Tyr Arg Val Leu Arg His Leu Ala Gin + 177 Pro 18. An anti-infection agent, characterized in that they contain, as active ingredient, a factor that stimulates granulocyte colonies in humans according to claim 17.