公开(公告)号：WO1984001150A1

公开(公告)日：1984-03-29

申请号：PCT/US1983001388

申请日：1983-09-13

Applicant: AMGEN

Inventor： AMGEN ,  SOUZA, Lawrence, M. ,  BOONE, Thomas, C.

IPC: C07C103/52

CPC classification number: C12N15/71 ,  A23K20/168 ,  A23K20/184 ,  C07K14/61 ,  C07K2319/02 ,  C12N15/69 ,  C12N15/70

Abstract: Novel polypeptides possessing biochemical and immunological properties of avian growth hormone are obtained by practice of recombinant DNA procedures. Novel DNA sequences are disclosed which are capable of directing the synthesis of such polypeptided in selected host microorganisms. In a preferred embodiment, a novel polypeptide with the properties of avian growth hormone of chicken species origins is produced as a result of bacterial expression of a novel plasmid, cGH-T21. This plasmid is harbored in transformed E.coli C600 cells deposited as A.T.C.C. 39182. Also disclosed is a DNA sequence capable of directing the synthesis of a novel polypeptide with the properties of avian growth hormone of turkey species origin.

Abstract translation: 通过实践重组DNA程序获得具有禽类生长激素的生物化学和免疫学特性的新型多肽。 公开了新的DNA序列，其能够指导这种多肽在选择的宿主微生物中的合成。 在优选的实施方案中，作为细菌表达新型质粒cGH-T21的结果产生具有鸡种来源的禽类生长激素性质的新型多肽。 该质粒载于转化大肠杆菌C600细胞中，淀粉为A.T.C.C. 39182.还公开了能够引导具有火鸡物种起源的禽类生长激素的性质的新型多肽的合成的DNA序列。

公开(公告)号：WO1986007387A1

公开(公告)日：1986-12-18

申请号：PCT/US1986001280

申请日：1986-06-13

Applicant: AMGEN ,  ABBOTT LABORATORIES

Inventor： AMGEN ,  ABBOTT LABORATORIES ,  SNITMAN, David, L. ,  STROUPE, Stephen, D.

IPC: C12Q01/68

CPC classification number: G01N33/54353 ,  C12Q1/6804 ,  C12Q1/6813 ,  C12Q1/705 ,  G01N33/532

Abstract: A method and a kit for the isolation and quantitative detection of a selected target nucleic acid sequence from solution employing two probes. A first probe is complementary to one portion of the target and is covalently attached to a first complexing agent (e.g., either an antigen or an antibody). The second probe is complementary to a different portion of the target and is associated with a reporter group. Following hybridization of the target and two probes in solution, a solid support coated with a second complexing agent (i.e., a corresponding antibody or antigen) capable of binding to the first complexing agent on the first probe is employed to immobilize the target-probe hybrid complex. A plurality of types of first probes may be used. Each type is attached to the same sort of complexing agent but each includes a nucleic acid sequence which is complementary to a different portion of the target.

Abstract translation: 用于使用两个探针从溶液中分离和定量检测所选靶核酸序列的方法和试剂盒。 第一个探针与靶的一部分互补，并且共价连接到第一络合剂（例如抗原或抗体）上。 第二个探针与目标的不同部分相互补充，并与报告者组相关联。 在溶液中杂交靶和两个探针之后，使用能够与第一探针上的第一络合剂结合的第二络合剂（即，相应的抗体或抗原）涂覆的固体支持物来固定靶 - 探针杂交体 复杂。 可以使用多种类型的第一探针。 每种类型都与相同种类的络合剂连接，但每种都包含与目标不同部分互补的核酸序列。

公开(公告)号：WO1985000831A1

公开(公告)日：1985-02-28

申请号：PCT/US1984001218

申请日：1984-08-01

Applicant: AMGEN

Inventor： AMGEN ,  BANKS, Allen, R. ,  PETERS, Mary, A. ,  SNITMAN, David, L.

IPC: C12P21/00

CPC classification number: C07K14/65 ,  A61K38/00 ,  C07K2319/00 ,  C07K2319/02 ,  C07K2319/75 ,  C12N15/70 ,  C12N15/72

Abstract: Manufacture of DNA sequences comprising structural genes coding for (1) a polypeptide having the amino acid sequence and properties of insulin-like growth factor I; for (2) a polypeptide having the amino acid sequence and properties of insulin-like growth factor II; and for (3) polypeptide analogs of insulin-like growth factor I and insulin-like growth factor II which differ from the naturally-occurring forms in terms of the identity and/or location of one or more amino acids, e.g., ADThr59 BD IGF-I and ADArg54Arg55 BD IGF-II.

公开(公告)号：WO1984004915A1

公开(公告)日：1984-12-20

申请号：PCT/US1984000834

申请日：1984-06-01

Applicant: AMGEN

Inventor： AMGEN ,  JONES, Theodore ,  RUDMAN, Christopher, G.

IPC: C07C103/52

CPC classification number: C07K14/61 ,  A61K38/00 ,  C07K5/1016 ,  Y10S930/12

Abstract: Novel synthetic peptides having primary structural homology to a continuous sequence of amino acid residues of human growth hormone in a region spanning positions thirty-two to forty-six ("hGH32-46", or "deletion peptide"). In preferred forms, peptides of the invention comprehend: duplicate portions (i.e., sequence fragments) of hGH32-46; stereochemical analogs and fragment analogs of hGH32-46 including one or more amino acid residues in D-isomeric configuration; and, "interspecies" analogs and fragment analogs of hGH32-46 including one or more non-homologous amino acid residues duplicating variant residues present in corresponding positions in corresponding regions of heterologous species growth hormones. Peptides of the invention are administered to mammals contemporaneously with exogenous insulin to generate hypoglycemic effects greater than available through administration of insulin alone. A presently preferred heptapeptide has the sequence NH2-Glu-Glu-Ala-Tyr-Ile-Pro-Lys-COOH, and has insulin-potentiating activity greater than hGH32-46.

公开(公告)号：WO1984002847A1

公开(公告)日：1984-08-02

申请号：PCT/US1984000063

申请日：1984-01-19

Applicant: AMGEN

Inventor： AMGEN ,  FOX, Gary, M. ,  HU, Sylvia, S.

IPC: A61K39/23

CPC classification number: C07K14/005 ,  A61K39/00 ,  C07K2319/00 ,  C07K2319/40 ,  C12N15/69 ,  C12N2750/14322 ,  C12Q1/701 ,  G01N33/56983

Abstract: Novel immunologically active polypeptide for use in anti-parvovirus vaccines through structural analysis and characterization of the parvovirus genome. In a preferred embodiment, microbial expression of polypetides is secured through use of DNA vectors comprising DNA sequences duplicative of porcine parvovirus (PPV) genomic DNA. Microbially expressed polypeptides as well as synthetic replicas of polypeptides coded for by DNA sequences of the invention exhibit immunological activity in, e.g., plate binding assays.

公开(公告)号：WO1984001787A1

公开(公告)日：1984-05-10

申请号：PCT/US1983001670

申请日：1983-10-27

Applicant: AMGEN

Inventor： AMGEN ,  ENSLEY, Burt, D.

IPC: C12P17/10

CPC classification number: C12P17/165 ,  C09B7/02 ,  C12N15/52 ,  Y10S435/849 ,  Y10S435/877

Abstract: Microbial synthesis of indigo dyestuff in indole-free media. Indigo production is preferably accomplished by genetic transformation of selected host cells having the capacity to produce and accumulate indole (either as a result of endogenous genomic capacity or genetic transformation) to incorporate the capacity for synthesis of an aromatic dioxygenase enzyme. Growth of transformed cells under suitable conditions facilitates aromatic dioxygenase enzyme catalyzed oxidative transformation of cellular indole, with consequent formation of indigo from the oxidized reaction products. In a highly preferred embodiment, E.coli cells having endogenous indole production capacity are transformed with a DNA expression vector comprising the structural gene for naphthalene dioxygenase, resulting in the microbial synthesis of isolatable quantities of indigo.

公开(公告)号：WO1987004461A1

公开(公告)日：1987-07-30

申请号：PCT/US1987000027

申请日：1987-01-07

Applicant: AMGEN

Inventor： AMGEN ,  STABINSKY, Yitzhak ,  ZUKOWSKI, Mark, M.

IPC: C12N09/56

CPC classification number: C12N9/54 ,  C12N15/90

Abstract: A mutated subtilisin suitable for admixture to washing compositions and exhibiting substantially improved stability over naturally occuring Bacillus serine proteases is prepared by expressing a modified gene encoding subtilisin in Bacillus subtilis. A preferred subtilisin analog product differs from wild-type Bacillus alkaline proteases by having any amino acid, and preferably serine, at position (218) in place of asparagine. The product is preferably produced in a strain of B. subtilis which is mutated to block synthesis of endogenous proteases. The method of replacing an Asn or Gly in an Asn-Gly sequence in order to improve pH and thermal stability may be applied to other sites in substilisin and to other proteins as well.

Abstract translation: 通过在枯草芽孢杆菌中表达编码枯草杆菌蛋白酶的修饰基因，制备适合于混合洗涤组合物并且显示出比天然存在的芽孢杆菌丝氨酸蛋白酶显着提高的稳定性的突变枯草杆菌蛋白酶。 优选的枯草杆菌蛋白酶类似产物不同于野生型碱性蛋白酶，通过在位置（218）代替天冬酰胺具有任何氨基酸，优选丝氨酸。 产物优选在枯草芽孢杆菌菌株中产生，其被突变以阻断内源性蛋白酶的合成。 为了改善pH和热稳定性，替换Asn-Gly序列中的Asn或Gly的方法也可以应用于替米多蛋白酶和其它蛋白质中的其它位点。

公开(公告)号：WO1987001129A1

公开(公告)日：1987-02-26

申请号：PCT/US1986001702

申请日：1986-08-15

Applicant: AMGEN

Inventor： AMGEN ,  FIESCHKO, John, C.

IPC: C12N05/00

CPC classification number: C07K14/005 ,  C12N1/18 ,  C12N2730/10122

Abstract: A method for culturing yeast capable of expressing an exogenous gene product wherein the yeast is cultured at about 25 C in a medium including balanced free amino acids at a concentration between about 20 g/l and about 160 g/l. Alternatively, yeast capable of expressing an exogenous gene product (e.g. the product of a gene introduced by the application of recombinant gene technology) may be introduced into a medium including a total carbon source concentration above but maintained as close to zero as possible, and cultured at about 25 C. A medium for fermentation of yeast capable of expressing an exogenous gene product includes balanced free amino acids at a concentration between about 20 g/l and about 160 g/l. The culture medium is preferably saturated with oxygen at least 30% of air saturation.

Abstract translation: 培养能够表达外源基因产物的酵母的方法，其中酵母在约25℃下在包含平衡游离氨基酸的培养基中培养，浓度为约20g / l至约160g / l。 或者，可以将能够表达外源基因产物的酵母（例如通过应用重组基因技术引入的基因的产物）引入到包含总碳源浓度高于但仍保持接近于零的培养基中的培养基中，并培养 在约25℃。 能够表达外源基因产物的酵母发酵培养基包括浓度在约20g / l至约160g / l之间的平衡游离氨基酸。 培养基优选用氧气饱和至少30％的空气饱和度。

公开(公告)号：WO1987001128A1

公开(公告)日：1987-02-26

申请号：PCT/US1986001704

申请日：1986-08-15

Applicant: AMGEN

Inventor： AMGEN ,  LEVINE, Howard, L. ,  LEVINE, Mark

IPC: C12N01/00

CPC classification number: C07K14/005 ,  C12N1/063 ,  C12N2730/10122

Abstract: A yeast cell lysis buffer for enhancing the recovery of subcellular particles including an aqueous solution of a nonionic detergent at a concentration within the range of about 0.5% to about 1.0% by volume at a temperature within a range from about 3 C to about 8 C. Triton X-100 is a preferred nonionic detergent for use in the lysis buffer. This lysis buffer may be employed in a method of extracting subcellular particles (e.g. HBsAg particles) from cells (e.g. genetically transformed yeast cells) which are broken open in the presence of the lysis buffer.

Abstract translation: 一种酵母细胞裂解缓冲液，其用于增强亚细胞颗粒的回收，所述亚细胞颗粒包括浓度在约0.5％至约1.0体积％范围内的浓度在约3℃至约1.0℃的范围内的非离子洗涤剂的水溶液 约8℃。 Triton X-100 R是用于裂解缓冲液的优选的非离子洗涤剂。 该裂解缓冲液可以用于从裂解缓冲液存在下断开的细胞（例如遗传转化的酵母细胞）中提取亚细胞颗粒（例如HBsAg颗粒）的方法。

公开(公告)号：WO1986007361A1

公开(公告)日：1986-12-18

申请号：PCT/US1986001281

申请日：1986-06-13

Applicant: AMGEN

Inventor： AMGEN ,  STABINSKY, Yitzhak

IPC: C07H01/02

CPC classification number: C07H21/00

Abstract: A method for the preparation of a 3' end functionalized polynucleotide. An amine-functionalized solid phase support is treated sequentially with an anhydride, then with an 1-hydroxylamine. A polynucleotide is chemically synthesized on the treated support and is subsequently cleaved therefrom by hydrolysis of the amide bonds. A polynucleotide having a 3' free primary amine is recovered for use in hybridization assays and other uses.