TW404951B - Effector proteins of rapamycin - Google Patents

Abstract

This invention comprises novel Rapamycin-FKBP12 binding proteins of mammalian origin for identification, design and synthesis of immunomodulatory, anti-restenosis or anti-tumor agents, as well as fragments of the proteins and the DNA, cDNA, antisense RNA and DNA segments corresponding to the proteins. This invention also comprises methods for isolating the proteins and therapeutic uses related to the proteins.

Description

Printed by the Central Labor Bureau of the Ministry of Economic Affairs and Consumer Cooperatives 404051 A7 _B7_ V. Description of the Invention (/) Effector protein of rapembranin The present invention is two proteins of rapamectin effector. Farewell • The novel threshing hormone-FKBP 12 binding protein of the present invention, which has no mammalian source, can be used to confirm, design and synthesize immune regulators, anti-restenosis or anti-plaque tumors. BACKGROUND OF THE INVENTION The macrocyclic antibiotics produced by Streptoayces hygroscopicus are first treated with anti-fungal properties. It adversely affects the production of fungi such as Candida albicans and Plastocystis. Ramen meal, its preparation and its antibacterial activity are described in (published on December 30, 1975, Surendra Sehgal et al.) US Patent No. 3929, 92. Marte 1, R. R., et al., Published in Canadian J 〇urna 1 〇f P hysio 1 ogica 1 P haraco 1 ogy, 5 5, 4 8-51 (19 7 7) in 1997. (Huisu's immunosuppressive properties on experimental sensitive encephalitis and psoralenitis. In 1989, Caine, RY, et al., In Lancet, 1989, Ho. 2., p. 227 and Morris, RE and Meiser · BM reported in Medical Science Research, 1989, No.17, p.609-10 respectively the effect of Lei 1te huixin on inhibiting allograft rejection in vivo, and many literatures then described the immunosuppressive and rejection inhibitory properties of rapamycin In addition, rapamycin has been used in clinical studies to suppress the rejection effect of human 酦 transplantation. Rapaviin alone (US Patent No. 4,885, 171) or combined with Pisibani (? 丨 with 68 [^ 1) Use (U.S. Patent No. 4,401,653) has been shown to have antitumor activity. RR Martel et al. [Can. J. Physiol, Pharecol. 55, 48 (1977)] shows that dipaxemin is effective (please listen first Read the notes on the back side # 栋 «'This page) This paper is based on the Chinese national standard (CNS > A4 size (21 0X297 mm) 404951 A7 B7 Consumption cooperation of employees of the Central Government Bureau of the Ministry of Economic Affairs · Du printed 5. Description of the invention (>) Sensitive brain spine of the experiment "Yan Qi, a chess form of multiple sclerosis; A supplementary rabitis model, a rheumatoid arthritis pattern; and effective in inhibiting the formation of IgE-like antibodies. The immunosuppressive effect of thalassin has been shown in PASEB 3,3411 (1989). Cyclosporin A) and other macrosulinone molecules such as FK-506 have also been shown to be effective as immunosuppressive agents and are therefore useful in preventing graft rejection [FASEB 3,341 1 (1989); FASEB 3,5256 (1989); RY Caine et al., Lancet 1183 (1978); and U.S. Patent No. 5, 100,899]. Ray meal has been shown to prevent or treat golden body lupus erythematosus [U.S. Patent No. 5,078,999], pneumonia [ U.S. Patent No. 5,080,899], Insulin-type diabetes [Fifth Int. Conf. InflaBB. Res. Assol, 121 (Abstract) · (1990)], and after vascular wounds] Muscle cell proliferation and vascular intimal hypertrophy [Morris, RJ H eart L ung T ransp 1 ant 11 (pt · 2): 19 7 (19 9 2)]. One of dipamuxin (acidified at positions 28 and 48) and a dialdehyde derivative have been shown to be water-soluble for use as an anti-emuth (U.S. Patent No. 4,316,885) and for the manufacture of rasmin Prodrugs (U.S. Patent No. 4,6 50,803). Recently, the position numbering convention of Rapahui Lei has changed; therefore, according to Chemical Abstracts nomenclature, the above esters are in the 31- and 42-positions, Meipu Patent No. 5,1 1 8,678 It shows rapamectin carbamate for immunosuppressive, anti-inflammatory, antifungal and antitumor agents. U.S. Patent No. 5,100,88 No. 3 shows a gasified ester of rapamycin. U.S. Patent Cases Please read the back note first-I-

IT This paper size is applicable to the Chinese National Car (CNS) M Washer (2 丨 0X297 mm) A7 B7 Printed by the Shell Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (Ή 5,118,677) Amine esters. U.S. Patent No. 5,13Θ, 3Θ7, amine esters of uranium and oxavidin. U.S. patent case 5,117,2Θ3, sulfonate and sulfonate esters of rapamycin, US patent Case No. 5,194,44 No. 7 shows sulfonal carbamate of rapifolin. U.S. Patent No. 5,1Θ0,899 (Caine) shows the use of thalamin and its derivatives and prodrugs to inhibit milk Methods of animal transplant rejection. Other chemotherapeutic agents listed for use with thalassin are Axatheth: azathi 〇prine, corticosteroids, sarsporin (and ascosporin A > And PK-506, or any of its cleavage products. Rapamycin produces an immunosuppressive effect by hindering intracellular signal transmission. Rapaparin shows non-calcium-dependent doubling transmission in stem fluorescent T cells and giant cells Cascade [Schreiber et al. (1992) Tetrahedron 48: 2545-2558]. Rappa It has been shown to bind to imaunophilins, which are members of the FK-506 binding protein (FKBP) family. In particular, when paravidin has been shown to bind to proteins, FKBP 12, FKBP 13, FKBP 25 [ Galat A. et al. (1992) Biochemistry 31 (8); 2427-2437 and Ferrera A, et al. (1992) Gene 113 (1): 125-127; Armistead and Harding, Ann: Reports in Med. CheB. 28: 207-215, 1993], and combined with FKBP 52 [WO 93/07269]. Rapafin can inhibit the proliferation of mitogen-induced T cells and B cells, and by several cytokines, including IL-2, IL -3, IL-4 and IL-6 (Review of Sehgal et al., Med. Research Rev. 14: 1-22, 1994) -5-Please read the notes first

I page order * • The size of the original paper is applicable to the Chinese National Standard (CNS) Α4 size < 210X297 mm) 404S51 Printed by the Bayer Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs «. 5. Description of the invention (10) Induced proliferation" > It is also possible to suppress the production of anti-I. Rapain purple has been shown to impede the activation of cytokine-induced i > 70si > 潋, which is now associated with the reduction of leiyouhuisu with cell cycle progression (Calvo et al .; Proc. Hatl. Acad. Sci, USA, 89: 757 and 7575, 1992; Chung et al., Cell 69: 1227-1236, 1992: Kuo et al, Nature 358 ^ 70-73, 1992; Price et al, Science 257: 973-977 , 1992) have differing abilities in protein synthesis. It also inhibits the activation of the cdk2 / cyclin E complex (Flanagan et al., Ann. NY Acad · Sci, in print; Flanagan et al., Mol. Cell biol, in print; Flanagan et al., J, Cell Bioche ·. 17A; 292 · 1993), the effect of thunderbolin was not directly bound to ρ7〇 * ί > stimulated by "I and cdk2 / cyclin E, but through the threshingin-FKBP complex in the ganglion kinase activity state Effect on the upstream components. The role of immunosuppressive cocoons, such as rapamycin, ascosporin, and KF506, is produced by the formation of a complex with eel proteins in other cells called iBBunophilins. When the combination of these immunosuppressive drugs and their individual immunophilins (iBilunophilins) can inhibit the cis-trans-peptidylprolyl isoform W (PPIase) activity of iBBunophilins, PPI as e-suppression is not sufficient to mediate the activity of immunosuppression (Review Ara stead and Harding, Annual Reports in Med. Chen, 28: 207-215: 1 993). Two rapamycin analogs of Diels Alder adducts, which have 4-phenyl-1,2 ,. 4-triazolidine-3,5-dione, Another 4-methyl-1,2,4-triazolin-3,5-dione, which binds to FKBP, has inhibited its PPIase activity, but it does not appear to be 任 In n HI i ^ nmi m n (谙 Read the intent on the back and then fill out this page) i The paper size is applicable to the Chinese national standard (CNS 棬 (2 丨 0X297 mm) 404951 A7 B7 Shellfish Consumer Cooperative, Central Bureau of the Ministry of Economic Affairs Printed 5. Description of the invention (() How can the immunosuppressive activity be detected. The phenyl-triazolindione diels-argel adduct with a high number of ears appears to be purple on the rapa emblem Competitive inhibitory effect on mitogen-stimulating DNA synthesis in mouse thymocyte proliferation (Ocain et al., Bioche .. Biophys. Res. CoBBun. 192: 1340, 1993). Recent evidence suggests that two equal parents Immuno-drug complexes, such as cyclophilin-chensporin A and FKBP-FK506, have a new function that can hinder fold transmission by acting on specific target proteins. Cyclophilin- The molecular targets of both cyclosporin A and FKBP-PK506 complexes, such as serine / residue-butyric acid phosphate that has been identified as Ca2 + / calcitonin dependent "I» Neuro (calcineurirOU. Liu et al., C e 1 1 6, 6, 8 0 7, 19 91; J. Liu and others, Biochistry 31, 3896, 1992; WM Flanagan et al. N. ature 352, 803, 1992; McCaffrey et al., J. Biol Chen. 268, 3747, 1993; KcCaffrey et al., Science 262: 750, 1993). The anti-splenic and immunosuppressive activities of rasmin were obtained through the use of fraxin, FK5 06 binding protein (FKBP) and At least one other third protein called rapamycin-like (TOR) is composed of a post-conjugation medium. The FKBPs family has been reviewed by Araistead and Harding (Annual Reports in Med. Chen, 28: 207-215; 1993) F. Lei, FKBP molecules in the antifungal activity of Huixin have been shown as FKBP 12 (Heitman et al., Science 253: 905-909: 1993). In milk cells, related FKBPs have been studied. Although two TOR proteins (TORI and TOR2) have been identified in yeast (Kunz et al .; Cell 73: 585 -596: 1 993), the target of rapamycin in human cells is still difficult to read first Note on the back

I Binding travel paper is again applicable to the Chinese National Dart Standard (CNS) A4 specification (210X297 mm) 404251 A7 B7 Printed by the Bayer Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Explanation of the invention (>). The carboxyl terminus of yeast T0R2 showed 20% identity with two protein peptones, the P110 subunit of phosphatidylin-based myozyme 3- 檄 W and one papillary yeast vacuolar picking protein VPS34 with PI 3K activity. However, J.Blenis et al. (AAI meeting, May. 1 993) have reported that the rapamycin-FKBP 12 post-compound does not directly mediate its effect on PDGF-excited cells via the P110, p85 PI3K complexes. Description of the invention The present invention is based on isolated, colonized and expressed proteins bound to the GST-FKBP 12-rapamycin post-complex. These proteins are isolated and prepared from the cell membrane of Molt 4T cell leukemia. Four types The size of the novel protein was estimated to be 125 ± 12 kDa, 148 ± 14kDa, 208 ± 15kDa, and 210 ± 20kDa by the PAGE shift method. The following and patent applications are shown as 125kDa, 148kDa, 208kDa, and 210kDa, respectively. The four peptones may also be referred to as effects in the following Application protein β The protein of the present invention can be used in screening analysis, such as plum inhibition analysis and binding analysis, to confirm the endogenous complexes and ligands and novel exogenous compounds (such as rapamycin) that regulate these functions. These proteins can also be used in analysis to confirm compounds for restenosis, immunomodulation with therapeutic effects, and as anti-sleepy sleep agents. The selection of the protein of the present invention not only allows the production of these proteins, but also provides a The basis for the development of related anti-sense therapeutics, using cDHAS colonies to produce antisense therapeutics with immunosuppressive activity (to combat transplant rejection, graft versus host disease, autoimmune disease , Such as wolf mill, myasthenia gravis, multiple sclerosis, rheumatoid arthritis, type 1 diabetes and inflammation, such as dry 8 This paper size applies to China National Sample Rate (CNS) Α4 specifications (210X297 mm) Please read Λ first Note-I- Binding Binding Printed by the Central Labor Bureau of the Ministry of Economic Affairs, Shellfish Consumer Cooperative _404951_B7__ 5. Description of the Invention (T), dermatitis, eczema, cortical leakage • Inflammation, Inflammation * asthma and uveitis), anti-restenosis and anti-tumor activity are included in the scope of the present invention. The protein of the present invention can be self-mammalian cells such as τ cell leukemia cell line, Molt 4 (ATCC 1582, Aierican Type Cell Culture) , 12301 Parklawn Drive, Rockvi 1 1 e, MD, USA, 20852) »Cell isolation of B-cell lymphoma cells, B JAB or normal human T cells. These tadpole milk cells can be used in a buffer containing protein gull inhibitor and reducing agent (2-ME) * such as low-purity buffer A (100 · Μ HEPES, PH7.5, 2Θ · Μ KC1, 1 · Μ EDTA, 0.4 · M PMSF and 2 · N-polyethanol (2-ME) were lysed. Nuclei and unruptured cells were removed by centrifugation at a temperature that reduced protein degradation to a minimum procedure. The membrane portion of the cells was then passed 100,000g high-speed centrifugation S to reduce or into small pieces. The cell membrane entrapment solution is dissolved in a buffer solution, such as buffer solution B (50 · (Tris, pH7.2, 1Θ0 · Μ NaCl, 2Θ · Μ KC1.Θ. 2βΚ PNSF, 1 · Η 2-ME CaCU. 2 · M figCU, 5 «g / · 1 aprotinin, leucine, gastrin A and analgesics), containing CHAPS0 (3-[(3- Azaminyl-propyl) dimethylamino] -propane sulfonic acid®; 12 · M) or Triton X100 (Polyoxan 4-Isooctyl Phenyl Ether) in a detergent. The dissolved film The protein can be separated from the debris by high-speed centrifugation at 100,000 g at a temperature that minimizes protein degradation. The supernatant containing the dissolved membrane protein is then reduced to a temperature at which protein degradation is minimized. When the inhibitor is present, it is pre-adsorbed with an affinity resin, such as glutathione resin. The resin is removed from the supernatant by centrifugation, and the supernatant is combined with rapamycin or rapamycin analogue to FKBP. , Such as GST-FKBP 12-Raypain, apply Chinese National Standard (CNS) Λ4 specification (2 丨 0X 297 mm) in reducing protein paper size ---------- k ----- ---- tr ------ Λ (Please read the precautions on the back before writing this page) 404951 A7 B7 Printed by Zhengong Consumer Cooperative, Central Rubbing Bureau, Ministry of Economic Affairs 5. Description of Invention (8) 霣 Degradation Cultivate at a minimum temperature. The dissolved membrane protein reacts with FKBP's combined rasamin or rapavin analog, such as GST-PKBP 12-rapaparin, and the mixture is then cultivated with affinity resin. In order to reduce the degradation of protein to a minimum temperature, rapamycin or rapatin analogs, FKBP conjugated proteins and protein binding compounds. Buffered fluorene, such as Buffered C (50bM Tris, pH 7.2, 100 · M NaCl, 20βΜ KC1, 0.2bM PMSF, ΙηΜ 2-ΜΕ or 10πΜ dithiothrease, 0.5βΜ CaCU · 0 -5βΜ MgCla, 5wg / Bl Aprotinin per, Leupeptin, Prionin A, and analgesics and 0.1% Tritonx 100) (Polyoxan 4-isooctylphenyl ether) After washing away most non-specific proteins, under denaturing (protein) conditions, such as a buffer containing the cleanest detergent (such as Laeili, Nature 227: 680, 197Θ with or without glycerin or along with the material La eiili buffer solution) to separate proteins from the resin, or under non-denaturing conditions such as those contained in affinity columns, such as glutamandum 2® when the eluting compounds elute the protein from the resin. These proteins can be separated by size using SDS polyacrylamide gel electrophoresis (SDS-PAGE). The present invention also includes the genomic DHA sequence of the aforementioned protein, and cDNA corresponding to the aforementioned peptone gene, anti-sense RNA, and antisense DNA sequence Yin j. The present invention also includes other mammalian proteins that are at least homologous or equal in function to the aforementioned proteins, and the DNA sequences of these homologous or equivalent proteins, and cDHA, antisense RNA and Meaning DHA sequence. For the purposes of this document and the scope of the patent application in Myanmar, the equivalent of the protein of the present invention is considered to be protein, protein segment and / or use of Chinese national standards (CNS specification (210 X 297 mm) Please read the previous note-Item Binding Printed by the Central Economic and Technical Bureau of the Ministry of Economic Affairs, Consumption Cooperative of the Employees, V. Invention Description (Gas) 1 I Proteins are basically similar, but not exactly the same in a truncated form. 1 Amino acid sequence. This equivalent exhibits rapamectin-FKBP conjugate binding properties and functions similar to the above-mentioned proteins. Therefore, in this specification and after the mine 1 I applied for a patent scope, corresponding to the present invention 125kDa, 148kDa , 208kDa and the first 1 218kDa protein pseudo-representation and it contains the same or equivalent protein, and read back 1 1 above 125kDa, 148kDa «2 081 ^ 3 and 2101 ^ 3 protein amino acid sequence Note 1 column I Segments and / or truncated forms that are similar but not identical in spring quality. $ 1 1 I These proteins or proteins are the same or equivalent Substances can be 'filled in 1] from different cell types using similar separation steps. Steps and / or the use of recombinant DNA methods to generate and write copies I may be directed to mutagenesis (si te di rected autagenes is) Η 1 1 technical modification. For example, the gene of the present invention can be designed to express one or more 11 proteins as a fusion protein, and the fusion partner can be used to confer 1 | advantages (such as HIS oligomer, immunoglobulin F c, Glutathione, S-shift 1 transposase $ FLAG, etc.). Mutations or truncations that produce soluble patterns can also be generated by site-directed mutagenesis and have advantages in isolation. 1 I The invention also includes retention The oligopeptide K segment that binds affinity but has less amino acid sequence than the active 1 1 protein, its truncated form and protein Η segment. The present invention also includes single strains and multiple strains specific to these proteins. Antibodies and their uses. Their uses include screening for novel drugs with immunomodulatory and / or 1 1 antitumor activity. Method and method of surveying the original compound and / or its substitute 1 Xie Wu contained in the biological sample of self-intake immunosuppressive__ individual method 〇 I use cDNA clones to generate immunosuppressive activity (transplantation Rejection,-Γ grafts against chronic diseases, autoimmune diseases such as lupus, myasthenia gravis, 1 | multiple sclerosis, rheumatoid arthritis, type 1 diabetes and inflammation such as psoriasis, 1 -11- 1 1 1 1 1 The size of this paper is applicable to China National Farming Corporation (CNS) A4 specification (210X297 mm) 404S51 ____B7_ 5. Description of the invention (I. ) Dermatitis, eczema, cortical leak, enteritis, pneumonia, asthma, and uveitis of the eye) and antisense treatments with anti-sleepiness activity (Mi Π igan et al., J. Med. Chen. 36: 1923-1936 , 1993), is also included in the present invention. The peptone of the present invention can also be produced by recombinant DH A technology, which is well known to those skilled in the art. Yi Di, the gene of the protein in question is also obtained by hydrolyzing the protein with protein 醑, such as lysine C, to obtain a part of the amino acid sequence, and then separating the resulting protein Η section by nicrobore HPLC, and then using fragment sequence analysis ( Matsudaira in A practicl Guide to Protein and Peptide Puri-ficatian for Microsequencing, Acadenic Press (San Diego, CA, 1989)). The measured sequences can be used to make oligonucleotide probes for direct screening of human cDNA libraries or to generate probes by polymerase reaction. Atlas can be produced from human T cells or cell lines, Molt 4, Jurkat, or other cells to obtain pupae. These colonies can be used to confirm additional colonies with additional sequences up to the protein gold gene, that is, a fully open reading frame, and are selected. Yin Jia, Consumers Cooperative of Central Bureau of Standards, Ministry of Economic Affairs (please read the note on the back before filling in this page) Some proteins in this technical field are known to be longer by a predicted length than the size of the peptone originally borrowed Encoded by the open reading framework ^ These protein sentences can represent the split products of the precursor protein translated from the complete open reading framework (such as IL-point) or the protein translated using the downstream start code (such as hepatitis B surface antigen) . In view of this, the cDN A-word in the scope of this article and the patent application refers to cDHA or a protein encoded by the gene with an open reading frame, which can encode a protein of the present invention or a protein bound or isolated as described above. Fragments, or in truncated form. In the complementary strategy, the gene of the protein of the present invention may be used by -12- this paper is of moderate size «β 家 标 牟 (CNS) A4 size (210X297 mm) 404851 at B7 staff of the Central Bureau of Standards of the Ministry of Economic Affairs Consumption Cooperation Du Yinzhuang 5. Description of the Invention (1) 1 I FKBP12 «RAPA is used as a colony trap to interact with the interactive yeast colony selection technique! I confirmed these techniques. These strategies can be combined to accelerate the confirmation of seedling expansion. 1 1 Relevant cDNAS clones encoding any one of the four proteins can also be expressed in vectors using the technology of the state of the art. In yeasts or rods, please first M 1 1 I virus-infected cells or mammals. Cells 0 can be isolated from PCJ 1 1 as described above | or a fusion partner (fusi 〇η pa Γ tner) can be used to confer separation advantage Note-meaning 1 I (such as Η IS oligomers * immunoglobulin Fc, glutathione Glycine s-transfer m etc.) 1 1 | Fusion protein is made. Mutations that produce a soluble form can also be generated by § > Λ from the location-directed mutagenesis method to obtain the superiority in separation. 〇 Writing s Packs I The use of cDNA clones such as this includes the production of recombinant proteins. Only 1 1, these recombinant proteins isolated from mammalian cells, or the corresponding natural egg 1 1 white matter 9 or its K segment (including oligomeric oligomers) have anti-eel for the production of these proteins 1 I quality. In other words, single or multiple anti-plaques are obtained by using recombinant proteins, 1 or the corresponding natural proteins isolated from white mammalian cells, or the segments (including and--containing proteins ( (Such as keyhole 1 I hemocyanin or bovine serum protein) conjugated peptide oligomers > and 1 1 induced. These antibodies can be used in natural protein 1 or white coli f isolated from mammalian cells. Purification of recombinant proteins isolated from yeast, or baculovirus-infected cells or lactation 1 Class I cells 1 or cell products. 1 1 The use of such cDNi colonies includes the disruption of recombinant proteins. Another 1 1 isolates white Recombinant protein of milk animal cells or Its corresponding natural egg-1 I white matter has a novel agent for screening immunomodulatory and / or antitumor activity-I > such as synthetic compound 9 natural product, exogenous or endogenous substrate 1 I method 〇 Natural products that may be screened may include, but are not limited to, 1 -13- φ 1 1 1 1 1 This paper size applies to China National Standard (CNS) M specifications (21 OX 297 mm) Central Bureau of Standards, Ministry of Economic Affairs Consumption cooperation by employees Du printed 404851 A7 B7 __ V. Description of the invention (u) Cell culture lysate, cell epithelium, plant extracts, and natural culture solution of true essence or bacteria. Examples of competitive binding analysis One of these proteins is linked to the matrix (covalently or non-covalently) and can be combined with a compound, natural product, cell lysate or cell supernatant, and a labeled rapamycin: FKBP complex The ability of compounds, natural products, exogenous or endogenous substrates to compete against inhibitory complexes or specific anti-membrane can be evaluated. Examples of methods include radiolabeling, fluorescent or chemiluminescent labeling, fusion proteins with FKBP such as luciferase, and binding to enzymes such as horseradish overgassing hydrazone, hyaluronic acid, ethanosine Esterase (ACHE) and so on. As an example of enzyme analysis, peptone is incubated with the substrate in the presence or absence of novel compounds such as synthetic compounds, natural products, and exogenous or endogenous substrates, and the enzyme activity of the protein can be evaluated. Assays of the original compounds and / or metabolites from biological samples taken from ingested immunosuppressive drugs can also be evaluated using these proteins. The invention includes a method for identifying a substance that can be used as an immunomodulator or antineoplastic agent, which method uses the following steps: a) The test substance is one of the four mammalian peptones of the present invention (125k Da, 148kDa, 208kDa, or 210kDa). ) Merged, peptone is bound to a solid support. b) Maintain the test substance and protein bound to the solid support of step (a) under conditions suitable for binding the test substance and protein. c) Determine whether binding of the test substance occurs in step (b). The invention also includes substances that can be used as immunomodulating or antitumor agents. The paper size is applicable to China National Cricket (CNS) A4 (210X297 mm) — I --- I- (Please read the back-$- (Item t fill in this page) 404951 Printed by the Central Ministry of Economic Affairs of the Central Government Bureau of Shellfish Consumer Cooperatives A7 B7 V. Description of the invention (6) The method of 1 I includes the following steps • ♦ 1 1 y I a) Combine the test substance with — A mammalian protein of the present invention, wherein the protein 1 is pseudo-bound to a solid support; 1 I b) under conditions suitable for binding the test substance to the protein, to maintain the test, please first bind the M 1 1 I substance to the protein The solid part of step (a) supports the earning. (Reading 1 1C) Determine whether the presence of the test substance is active in mammalian proteins. The above note-1 I The present invention also includes a biological sample for detection, when Dangdang Baihuisu, Raibai $ 1 item-| Huimin analogues or rapamycin metabolites are combined with a FKBP, and then combined to 1 A method of one of the four listed proteins of the present invention, the method comprising steps: a book binding page 1 a) combining biological samples with a FKBP to form a first mixture, such as rapamycin 1 I, dipavin Analogs or rapamycin metabolites are present in biological samples. The 1 1 mixture contains a rapamycin: FKBP complex, thalassin extracts: 1 I FKBP post-complex, or rapamycin Metabolites: a mixture of FKBP complexes; order | b) 葙 by adding a protein of the present invention to the first mixture to make 1 1 to obtain a second mixture 9 protein pseudo-bound to a solid branch; 1 IC ) In combination suitable for rapamycin: FKBP complex, rapamycin is similar to 1 I: FKBP post-compound t or rapamycin metabolite: FKBP complex ( If the existence of \ A under these conditions), maintain the second mixture of 1 I and 1 I of the protein of the present invention in step (b) and d) determine rapamycin: FKBP complex in step (c) Fructoplasmin 1 analogue · FKBP complex * or rapavinin metabolite: whether FKBP complex 1 I binds to protein 0- 1 1 Also included in this issue is the use of cDNA colony The body produces antisense treatment 1 1 〇 This can be achieved by current technology, such as described in Milligan et al. J. Med. 1 I _ 1 5 _ 4 1 1 1 1 1 This paper size applies Chinese National Standard (CNS) A4 Specifications (210X297 mm) _951-Seal of the Shellfish Consumer Cooperative of the Central Government Bureau of the Ministry of Economic Affairs. 5. Description of the invention ((屮) 1 1 Ch eo. 36: 14: 1924- 1936. Completed for use as a guide and continuation. The special t antisense RNA and DNA images of Lishen's range include derived white papers with four inventions (125kDa, 148kDa, 208kDa or 210kDa). Please 1 | Skeleton II · Coding cDNA Select these RNA and DNA strands of the 鳢 or use the first read 1 I The person who modified the skeleton 〇 Such RHA and DNA skeleton modifiers are described on the back Xy 1 1 Mi 1 1 iga η et al. J. M e < j * Ch en. 36: 14: 1924- 2936 〇 Narrated by the note-1 1 (Μ ί 1 1 i ga η et al., J • Ned. Che fll * 36 : 14: 1 924- 1936) 1 2 '| 1 I Anti-sense compounds produced by current stage technology are used in sacral immune response packs | 1 Therefore they are used for transplant rejection »such as kidney f heart 臓 liver 臓 liver * bone% Posts on this page * Pancreatic crest (islet cells) »Corneal, small intestine and dermal xenograft $ and 1 I Heart valve xenograft treatment or suppression; for white body immune disease f such as wolf 1 1 sore% wind m Inflammation »Diabetes, Myasthenia gravis and multiple sclerosis I and inflammation t such as dry moss, dermatitis, eczema 1 cortical leak, win inflammation and eye fixation I uvitis 〇 The antisense molecule of the present invention also has antitumor 1 tumor 1 antifungal activity 1 and antiproliferative activity 0 The compound of the present invention is therefore also 1 I can be used to treat lumps of adult T cell leukemia / lymphoma% true 1 1 And hyperproliferative vascular diseases »such as restenosis and arteriosclerosis1 * so% The present invention also includes the treatment of mammals, preferably 9 humans, as described above 1 I diseases and conditions, the method includes administering effective 1 to 1 mammals in need Invention of a related antisense therapeutic agent. 1 When applied to treat or inhibit the above-mentioned disease states 9 The anti-sense of the present invention 1 | The child can be taken to mammals »Intestinal cost extra * Intra-selection > Intrabronchial 9 Meridian-1 Skin t Local 9 Intravaginal or intrarectal Administration 〇1 I When antisense and antisense molecules of the present invention are used as immunosuppressive or anti-inflammatory agents, they can be used with — 1 -16- 1 1 1 1 1 This paper size is applicable to Chinese national standards (CNS > A4 « L 棬 (210X 297mm) ^ 04851 A7 B7 Printed by the Central Government Bureau of the Ministry of Economic Affairs, Printed by the Shellfish Consumer Cooperative, V. Description of the Invention 05) 1 | or more other combinations of immunologically controlled drugs are expected. Other I immunomodulators include 9 but are not limited to axabinil (a ZB th 1 0 ρ Γ i ne) 1 1 «$ corticosteroids, such as prednisolone (predn is ο η e) and 6 A Prednisone please read 1 1 pine (eethy 1 p redn is ο 1 one), cyclic m amine • rapamycin 1 spore 1 I prime A, FK-506. 0KT " 3, and ATG〇 borrow Combining the complex of the present invention with reading back 1 I side I such other drugs or agents to induce immunosuppression or treatment of inflammatory conditions Note 1 1% A smaller amount of each agent is required to achieve the desired effect. Event 1 I Therapy foundation was established by Stepko vski, and the results show that the combined use of sub-therapeutic medicament I and thalassin and cinsporin A effectively prolonged the survival time of heart palpitation xenograft. C Tran SP 1 antat ion Pr 0C .23: 507 (1991)] 〇1 I Therapy using these antisense compounds usually starts with a smaller dose than the compound's best 1 1 1 and then increases the dose until the best 1 I effect is achieved under the circumstances. The precise dose is determined by formulating 1 based on the experience of the treatment master. — * In general. The antisense compounds of the present invention most hope to achieve effective results at a speed of 11 to 1, but have not produced any harmful or harmful side effects. 1 1 I Therapeutic value of the antisense compounds in accordance with the above% The present invention also includes a pharmaceutical group of derivatized 1 A white containing coding for the 125kDa 9 148kDa t 208kDa and 210kDa proteins of the present invention 1 I cDKA antisense RHA and antisense DHA compounds 1 1 Finished. 1 \ The present invention also includes the following methods for isolating the protein of the present invention and the isolated 1 I peptone:-1 * 1 A method for separating protein from mammalian cells »It includes steps: 1 I 1. Culture and collection of box milk Cell-like cells as described above »The cells can be used in 1 I -17--1 1 1 1 1 sizes of this paper. Chinese National Standards (CNS) A4 (210X297 mm) 404951 B7 Central Bureau of Standards, Ministry of Economic Affairs Consumption Cooperative Seal 5. Description of the Invention (^) 1 | Can be T cell source (such as T cell lymphoma, leukemia, normal human TI cells), B cell source (such as EBV transformed B cells, normal human B cells) 1 f Giant cell ginseng or other cellular sources sensitive to rapamycin. The cells may be used for work soon after collection or may be frozen before the work is performed. 1 I germanium is stored 1 if it is granulated. Cold * cells may be stored in dry ice and ethyl alcohol bath before use. > Store frozen at -70--80Ό. Note for this training and collection of V7-1 The step of I cells may be regarded as the first step of this method or only %% of this method-I Preliminary steps 0 and I 2. Cells are reducing protein degradation to a minimum Temperature (eg 41)% of this %% loading I «in a buffer containing (such as HEPES, Tris, f > H7.5), low 迩 (such as 10 1 I 50 Bl Vi N aC mm), chelating agent (Such as 1 ~ 2 · M EDTA), a buffer of protein W 1 inhibitor (such as 0.4 PMSF) and an original agent (such as 2 · M 2-ME or 1 ~ 20 1 IN dithiothreitol) Medium lysis, 晡 milk cells may be treated by any of the methods that can produce cytolysis, including sonic lysis and homogenization. 1 1 Plasma vessels are understandable. 0 3. Unbroken cells and nuclear nuclei By centrifugation at a temperature that reduces protein degradation to a minimum of 1 1 (such as 41C), the lysate is removed first, and centrifugation at 9 such as 1600g for ten minutes has been sufficient to remove undisrupted cells and Nucleus 0 this step, Mandatory, and then provides the following 11 steps a more Qing Che of articles square 14, while after the membrane fraction after the first row to clear the lysates in the as by 1 I of a high-speed centrifugation was added to concentrated. An example of a concentrated spring is high-speed separation at 100,000 g for 1 to 1.5 hours. 〇1 | 5. Then membrane proteins (such as transmembrane proteins, membrane proteins and binding proteins) 1 I -18 " 1 1 1 1 1 This paper size is in accordance with China Standard for Household Standards (CNS) 8-4 (210X297 mm). Printed by WC Industrial Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs ^ 04351 A7 __B7__ 5. Explanation of the invention (Γ)) is dissolved. This can be done by incubating the pellets from step 4 in a detergent containing soluble protein but not denaturing it, a buffer (such as 20 ~ 50bM Tris or HEPES, pH7.2), salt (such as 100 ~ 200bM HaCl + 20nM KC1), reducing agent (such as 1 ~ 2bM 2-ME or 1 ~ 20 · M dithiothrease), protein plum inhibitor (such as Θ.2 · Μ PMSF, 5 / ug / il aprotinin, Leupeptin, gastrin A and analgesics), divalent cations (such as 0 ~ 5 · M CaCU, 0 ~ 5 · M MCU). Examples of detergents used in this step are CHAPS0 (3-[(3-aminoaminopropyl) dimethylaeroline] -1-propanesulfonate) or Tritonxl00 (polyethylene 4-isooctylphenyl ether) ). After thinking about this, the mixture contains dissolved membrane proteins and undissolved cell debris. 6. The solubilized membrane protein is separated from the undissolved cell debris, such as at a temperature that reduces protein degradation to a minimum (such as 4t: > by high speed _ centrifugation (such as 100,000g 1 ~ 1.5 hours). 7. Supernatant containing dissolved membrane protein and an affinity resin at a temperature that allows protein to be directly adsorbed on the affinity resin and reduces protein degradation to a minimum (eg 4t: > and time, in a buffer containing one Agent (such as 20 ~ 50nM.Tris or HEPES, pH 7.2), salt (such as 100 ~ 200bM HaC, 20bM KC1), burial agent (such as 1 ~ 2bM 2-ME or 10 ~ 20bM dithiothreitol), chamber White plum inhibitors (such as 0.2nM PMSF, 5 / zg / nl aprotinin, leupeptin, pepsin a and analgesics), divalent cations (such as 0 ~ 5 mM CaCl2, 0 ~ 5nM MgCU) Incubate in buffer. 8. Resin is removed from the supernatant by centrifugation at a temperature that minimizes protein degradation to a minimum (eg 4¾). 9. The supernatant is then combined with a fusion protein complexed to FKBP 12 + protein- 19- This paper is suitable for 8 | National standard (0 milk) 8 4 specifications (210 fathers 297 males) ---------------, tr ----- -(Please read ii on the back, Yi Shi Yan, and then 4 '· write this page) ^ 04951 A / B7 Printed by the Central Bureau of Standards, Ministry of Economic Affairs, Shellfish Consumer Cooperatives Cultivar or thalassin analog (ICt * in LAF < 500 μM), the fusion protein strengthens the separation of the target protein and thus allows the fusion protein to allow the effector protein to bind to the fusion FKBP protein ： The combination of rapamycin or analog complex and reduce protein degradation to the minimum temperature and time (such as 4¾ 1 ~ 2 hours) under a buffer containing (such as 20 ~ 50bM Tris or HEPES, pH7.2 ), Salt (such as 100 ~ 200bM NaC and 20bM KC1), reducing agent (such as 1 ~ 2bM 2-ME or 1 ~ 20bM dithiothreitol), proteinase inhibitor (such as 0.2 · M PMSF, 5wg / »l Aprotin, Leupeptin, Stomach "Astatin A and anti-pain_), a bivalent limit ion (such as 0 ~ 5 · M CaCU, 0 ~ 5 · Η MgCU) in a buffer solution with an affinity resin or affinity Sex column, such as GST-FKBP12, histidine oligomer-FKBP12, biotin-FKBP12, etc. 10. Effector protein and fusion FKBP protein: Thunder meal emblem Step 9 mixture of the admixture at a temperature and time (such as 4C and 0.5 ~ 2 hours) that allows effector proteins and fusion FKBP proteins: rapatin or the like to bind to the affinity resin and reduce protein degradation to a minimum Public affinity resin cultivation. 11. Most non-specific proteins dissociate the binding of non-specific proteins, but do not 'dissociate' the desired protein from the RAPA-FKBP post-complex buffer wash, such as a buffer containing a buffer (such as 20 ~ 50bM Tr is or HEPES, pH7.2), salt (such as 100 ~ 1000bM HaCl + 20 «iM KC1), reducing agent (such as 1 ~ 2 * M 2-ME or 10 ~ 20bM dithiothreitol), protein W inhibitor (such as 0.2niM PMSF, 5 «g / ul aprotinin, leucopeptide purple, peroxin A and analgesics, divalent cations (such as 0 ~ 5 bM CaCU, 0 ~ 5bM MgCl3 -2 0-please first Notes to the front page · I- The size of the bound paper is in accordance with the Chinese National Standard (CNS) A4 (210X297 mm). Printed by the Shellfish Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs 404951 a7 ___B7_ 5. Description of the invention (W)) And a detergent that can dissociate non-specific peptone binding, but does not dissociate four proteins with RAPA-fused FKBP protein, such as Triton X100 (Polyoxetyl 4-Isooctyl ») buffer. 12. Effector protein fusion FKBP protein: Rapain complex is eluted from the resin with a suitable buffer, such as a buffer containing sufficient detergent to dissociate it from the resin (such as with or without glycerol or dye Laeiili buffer, Laemli, Nature 227: 680, 1970), or an appropriate elution compound such as glutathione, histidine in an affinity column. 13. Effector proteins can then be separated by size. This can be done by any method of separating proteins by size, including, but not limited to, polyacrylamide gels, swimming methods and size-flick column chromatography. It may also be used to compare male and female proteins by a control step, which is a pseudo-replacement of step 9 rapamycin or rapamycin analog (IC0 in LAF < 500n »O step of buffer solution). Thinking, this step can be used to more easily distinguish the protein bound to Dioscorerin: FKBP after-complex. The peptone of the present invention can also be cracked by recombinant DHA technology familiar to those skilled in the art. That is, the desired The gene of the protein can be obtained by digesting a part of the amino acid of the protein with an appropriate endopeptidase, such as lysine C enzyme, and separating the obtained protein fragment by nicrobore HPLC and further reading the sequence (M atsudairain AP ractica 1 G uidet ο Ρ 〇r * tei η and PeptίdePurificatiοn for Microsequencing, Acadeinic PressSan Diego, CA 1989) for cloning. the determined sequence may be used to manufacture may be used to screen human cDNA made Hall FIG Wei, such as human T cells, Μ 0 Η 4, Jurkat's Oligonucleotide Probes (Sabbr οk, Fritsch, and -21- This paper size is applicable to Zhonggu family promotion (CNS M4 grid (210X297 mm) --- ---.-- 丨: Install -------- Order ------ line (please read the precautions on the back before filling this page) V. 404951 A7 B7 Central Bureau of Economic Affairs Shellfish Consumer Cooperative's Neem invention description (β) Haniatas, Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press, 1989). These breeding feeds can be used to identify other clones containing additional sequences until the protein is complete and the gene is removed. Breeding (Sabr οk, Fritsch · and Maniatas, Μ 1 ο 1 ecu 1 ar Cloning, A Laboratiory Manual, Cold Spring Harbor Press, 1989). In a complementary strategy, genes may use FKBP12: RAPA as Breeding traps are identified by interacting yeast S colonization techniques (Chien et al., Proc. Natl. Acad. Sci. 88: 9578-9582 1991). These strategies can also be combined to accelerate the identification of colony plaques Relevant cDNAs can also be expressed in E. coli, yeast or baculovirus-infected cells or mammalian cells using expression vectors of current technology. Isolation of cDNA or cDNA as described above can be made in the form of a fusion protein (such as HIS oligomers, immunoglobulin Fc, glutathione S-transferases, etc.) in a fusion protein that confers excellent adhesion. Mutations that cause soluble forms can also be generated by site-directed mutagenesis, and have the advantage of heterogeneity. Articles of rats, rats, monkeys, dogs and other mammals can be obtained by similar steps. In addition, when the selective colony of human proteins is isolated, the colony can also be used to screen other mammalian homologues. This equivalent line can also be used to develop binding analysis and to establish a large number of compounds that can be screened for endogenous ligands. Screening analysis of exogenous preparations to confirm their activity and immune activity. Compounds with such immunomodulatory activity, endogenous ligands, endogenous ligands and exogenous ligands can be used to regulate the immune response and so there is -22- (Please read the precautions on the back before filling out this (Page)-Installed. -----! · Binding line The paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) 404951 at _B7 _ V. Description of the invention (> 丨) Used for transplantation rejection effects such as Treatment or suppression of kidney, heart, liver ridge, lung, epiphysis, pancreas (islet cells), sclera, small intestine, and skin graft xenograft and heart valve transplantation; autoimmune diseases such as lupus, rheumatoid arthritis, diabetes , Treatment or suppression of myasthenia gravis, and multiple sclerosis; and inflammation, such as dry addiction, dermatitis, eczema, cortical leakage, enteritis, and uveitis. The above-mentioned compounds, endogenous ligands and exogenous ligands can also have anti-tumor pain, anti-fungal activity, and anti-proliferative activity. The compounds of the present invention can therefore be used for the treatment of adult tumorous T-cell leukemia / lymphoma, fungi of hard tumors. Qi stain, and hyperproliferative vascular diseases such as restenosis and arteriosclerosis. Example 1 The protein of the present invention was isolated using a fusion protein of glutathione S-transfer SS-FK506 binding protein 'white matter 12 (GST-FKBP). GST-FKBP is produced by recombinant E. coli containing pGEX-FKBP. Cells were cultured, induced by IPIG and used as taught in D, B. S bithand K. S. Johns ο, Gene 67, 31 · 1988 and K · L. Guan and JE Dixο η, Andl. Biocheb. Standard techniques of 192,262, 1991 separate the fusion proteins. The solution containing glutathione and GST-FKBP12 was exchanged 5 times using a Centricon 10 filter (Amicon) to remove glutathione and replace the buffer. Printed by the Shellfish Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the note ^^ on the back before filling this page)

Molt 4 cells (1X10®) were cultured in a standard culture solution (containing 100U / β1 penicillin · 100 «g / nl L-glutamine, 10% FCS RPMI 1 640). Cells were harvested and washed 3 times with PBS (50dM phosphate buffer, pH7.0, 150bM NaCl), fresh frozen in a dry ice ethanol bath, and stored at -80t :. On ice, the cells are thawed and buffered in 5B1-23- This paper scale applies Chinese national standard (CNS > A4 this grid (2 丨 OX297 mm) 404951 A7 B7_ V. Description of the invention ((Please read first Note on the back, please fill in this page) Printing solution A (1Θ · Μ HEPES, pH7.5 · 20 · Μ KCl.laN EDTA, 6.4bM PMSF and 2bM 2-ME) The dounce homogenizer with type B ground birch was dissolved. After the debris was removed by centrifugation at 1600g for 10 minutes, the membrane part was concentrated by centrifugation at 100,000g (1 hour), and 100,000g pellets were contained in 3bI containing 12bMCHAPS0. Buffer B (50βΜ Tris. PH 7.2, 100bM NaCl, 20eM KC1, 0.2n- MPMSF, 1 · M 2-ME, 2nM CaCla, 2 · M MgCU, 5a g / nl aprotinin, leucine, gastric enzyme Inhibin A and analgesics) were incubated at 4 t! For 2 hours. The dissolved membrane protein was separated from the incubation by centrifugation at 100,000 g. The supernatant was swollen by 0.4 · 1 glutathione in buffer B After the glycine agarose resin was first adsorbed for 3-18 hours, the supernatant and the compound Raynin-GST-FKBP12 (from 660 wg GST-FKBP + 60 / zg ΚΑΡΑ were incubated in buffer B, 4 * 0 1 Formed in ~ 2 hours) Incubate at the bottom for 2 hours. Then the supernatant is incubated with 100 «丨 glutathione resin U: 1 buffer B) for 2 hours at 4t. Non-specific proteins are buffered with buffer C (buffer B) + 0.1% T rit η X 1 0 0) and wash 5 times. The protein was eluted from the resin in Laeeli buffer by incubating at 95 ° C for 3 minutes and thoroughly centrifuged. The protein was extracted by 7% SDS-PAGE. Size separation. The silver staining method should be followed. The concentration of the four plaques corresponding to proteins of molecular weight 210kDa, 208kDa, 148kDa, and 125kDa in samples containing RAPA + GST-FKBP12 was higher than that in samples containing only GST-FKBP.

An analysis of mitogen-stimulated thymocyte proliferation known as LAF can be inhibited by rapamycin or an analogue such as noroxygen rapamycin and indicates the relative activity of rapatin analogs in immunosuppression, the same Protein utilization and immunosuppressive analogues, nostril rapamectin (Table 1) compounded GST -24 · This paper size applies the Chinese National Standard (CNS) A4 specification (210 × 297 mm) year ^ amendment 40495J., Sa丄 止 才 攱 charge Α7 Β7 V. Description of the invention (8) (Please read the precautions on the back before filling out this page) -FKBP is separated, Diels Adele adduct binds to FKBP12 and inhibits FOP1 2 PPIase activity, but does not show detectable immunosuppressive activity, so it does not bind to the target of rapamycin. Use these two compounds complexed with GST-FKBP12 in a similar separation step (that is, replace rapatin : GST-FKBP12), yielding a 210 kDa protein at a background concentration (without rapamycin) (Table 1). FK506, an immunosuppressive compound, has at least certain functions via the FK506-FKBP complex and calcium button binding mediators. * FK506 complexes with GST-FKBP in similar steps only produces 210 kDa protein at background concentrations (Table 1). Table 1 Comparison of rapamycin analogue-FKBP12 complex and 210kDa protein binding

Compound_ 210kDa LAF PPlaseiKi) RAPA + + + 6nM 0. 12nM demethoxyrapamin (immunosuppressive analogue) + + + 58nM 4.4nil Dielsader adduct (phenyl) soil > 1000nM 12nM Diels Adele Adduct (Methyl) + 1000nM 12nM FK506 3Nra * 0.4nM None (FKBP) (* Different mechanism of action)

The action of rapamycin, its analogues and other agents such as FK 5 06 suppresses the immune system, and they perform this kind of action by binding to the respective hippoproteins. This binding inhibits the PPI a se activity of the receptor protein However, inhibiting PPIase activity alone in combination with rapamycin or its analogs does not only cause immune suppression, it is indeed due to two rapamycin analogs (Diels Alder adduct in the table) adducts)) bound to FKBP by -25-

This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210X2 blade public director T 404951 Π year 丨 called B7) V. Description of the invention (smart) Body protein and PPlase that inhibits FKBP * But it cannot prove any detectable immunosuppressive activity, This has been resolved in paragraph 2 on page 6 of the instruction manual (paragraph 3 on page 3 of the English manual). Thymus cell proliferation assay is called LAF. An assay is commonly used to indicate the relative immunosuppressive activity of some compounds, such as thunder The higher the level of palmitoxin analog, the lower the immunosuppressive activity of the compound. The data indicate that the 210kDa protein was separated by two different methods (the first was separated by GST-FKBP12 and RADA, and the second was separated by GST-FKBP and Demethoxyrapamycin separation, the first two rows of the table) shows low PPUse activity and more immunosuppressive activity compared with two Dielsader adducts and FK506, in the LAF assay These two Diels Adel adducts and FK506 demonstrate lower immunosuppressive activity (> 100 nM, and the known immunosuppressive tincture FK506 is 3 nM, and the 210kD protein separated by the two methods described above is divided into 6nM and 58nM), and more PPIase activity (more than 10 times). It is known that rapamycin must be combined with a member of the FKBP family to mediate its effect. In order to prove the binding of the protein of the present invention to the complex RAPA-GST-FKBP ( Please read the notes on the back before filling in this page) Non-individual use in combination with rapamycin or FKBP12 has been amended to separate the steps of consumer cooperation of the Intellectual Property Bureau of the Ministry of Economic Affairs. This amendment includes the use of (1) a mine Palmitoin-42-biotin glycinate replaces rapatin (all show equivalent immunosuppressive activity in LAF analysis), (2) no exogenous FKBP and (3) —Stripar Davidin (Strepatavidi) A binding resin instead of glutathione. Resin composition * Only the background concentration of 2101 cDa protein was separated using this modified separation step. The 210 kDa protein was using the GST-FKBP12-rapamycin complex by reference-Haipak ( Ficoll-Hygauue) and T cell column purified normal human T lymphocytes and BJAB cells (B-cell lymphoma cells) and their isolation. The results of some amino acid composition analysis are listed in Table 2 » Percentage union -26- This paper size is in accordance with Chinese National Standard (CNS) A4 specification (210X297mm). V. Description of the invention (reserved component name α in the spike table except 18! Spikes and 蒱 reactions Min 芊 Sharpness Η 〇. /. ς Λ 9.38 11.09 1 Asp / Asn 12.06 12.47076 0.02344 .0.05142 () .30 2 Tfir .13. () 5 2.92898 (). () () () 00 0.00985 0.068 3 Ser 13.78 6.43968 (). () () ( X) () 0.01995 0.15 15.68. 4 Glu / Gln 16.87 25.47273 0.00000 0.05285 0.59 Prp 18.24 0.1 ^ 1 5 Gly 22.35 21.50384 0. () () (X) () 0.04645 0.44 22.90 6 Ala 23.73 16.69160 0.0 () (X) 0 0.03113 0.36 26.06 28.81 7 Val • 29.39 4.83196 0.00000 0.00605 0.11 'Met 32.28 8 lie 34.10 3.00560 0.2326 0.00782 0.0699 9 Leu 35.09 5.73202 0.02331 0.01372 0.1383 10 nLeu 36.27 20.48232 0.02174 Q.042S6 0.4453 11 Tyr 38,33 1.44792 0.00 0.02 40.05 1.25017 0.02703 0.00187 0.0338 13 His 47.79 1.50905 0.02553 0.00580 0.0385 14 51.80 12.66136 0.00000 0.01960 0.000 () 15 Lys 53.34 9.90767 0.0228 3 0.02274 0.2262 No. 146.553645 0.33436 144.29 Undecided * 7-Real-time consumer cooperation in the sample rate bureau of the Ministry of Economic Affairs Du printed 404951__J; __V. Description of the invention (虬) Example 2 210kDa (210 ± 20kDa) protein of the present invention Isolation was performed from 4X104 Molt4 cells using the affinity matrix method as previously described. The bound protein was eluted from the affinity matrix with lx LAEMLI buffer (0.0625: 1 ^ 8-HC1, pH 6.8, 2% SDS, 0.37M / 3-methylacetone) without glycerol and dye. Centrifuge 3 times using centricon 100 (Anicon, Beverly, MA) connection and centrifuge, the first two at temperature 4¾ • the second at 18¾. The concentrated sample was eluted from the centricon 1Θ0 filter by incubating the same volume of 2x Laenli buffer without glycerol and dye for 2 hours at room temperature. The first 2x and third 2x Laenli buffers were used. Performed on a 1.5 πβ thick 7% polyalbumin gel (38: 1)? Six £ 6 and separated. The protein was transferred to a polyvinylidene difluoride, PVDF (Biorad, HerculeSi (A), PVDF (Biorad, HerculeSi (A)) at 50bAbps, 4% overnight at 0.01% 13 / glycine buffer solution (Biorad) containing 0.037% 5.05. ). PVDF protein stained with amine black (Biorad) in 10% acetic acid · 2% acetic acid • and cut off the appropriate plaque, washed with PBS and water and stored frozen. Sequence determination of protein on PVDF membrane (about 3w g) In situ hydrolysis with trypsin according to the modified method described by J. Fernanez et at, (Anal. Biochen. 201: 255-64, 1 992). In a short time, PVDF was cut into Ibb2 broken K, first moistened, protein Hydrolysis in 100bM Tris-HCl containing 10% acetamidine and 1% at 0.2 «8: Trytonin reduced triton (CalBiochen), pH buffer at 371 for 6 hours, then 0.2 / zg trypsin was added and Incubate all night. The M section is buffered by sonication from the membrane and contains the K section buffer. -28- (Please read the note on the back before filling in this page) This paper size applies Chinese National Standard (CNS) Α4 Specifications (210X297 cm) Printed by the Central Sample Rate Real Consumer Cooperative of the Ministry of Economic Affairs ^ 04951 at B7 V. Description of the invention (3) Centrifugal separation by small tubes. Extract again 2 times (ie add 50 w 1 buffer to the membrane, sonicate, centrifuge in a small tube and combine the solution with the original buffer containing the eluted fragments). Sample (140-145 / i 1 ) By using a Vydac C182.1bbX 150ea reversed-phase column in a W. Lane et al., (J. Protein Che »., 10 (2): 151-60, 1991) 40-pole matrix detection previously described The narrow-bore HP LC was separated on a Hewlett Packard HPLC 1090 of the detector. Multiple fractions were collected and the absorbance was measured at multiple wavelengths (210, 277, and 292η ·). According to resolution, symmetry, and UV absorption and spectrum (210 mu, 277η and 292ηβ) Select the best part for sequence determination. The 5% better part is by oatrix assisted laser desorption tine of flight nass on a Finnigan laser device spectrometry (MALDE-TOF-MS) analysis of its homogeneity (hoBogeneity) and fragment length. The best part selected was automatically performed on the Applied Biosystees 477A protein sequencer by using a 撖 cassette and the chemical cycle recommended by the manufacturer. Ed »a η degradation order . The sequence comparison was performed using the Intelligenetics suite (Intelligeneitcs, (A). The sequence was determined using the above method to determine the 210 kDa (210 ± 28 kDa) protein of the present invention containing a peptide peptide segment, of which four have the following amino acid sequences: a) ILLKIEHR; B) LIRPYMEP ILK; -29- — -HI n nnm * equipment — I— * 1 order one * —, III — line 'I (please read the precautions on the back before filling this page) This paper size applies to Chinese national standards (CNS > A4 specifications (210X297 mm), the Ministry of Economic Affairs, the Chinese Communist Party, the rate of f, the cost of work, and the cost of the cooperative, and the A7 _B7 _V. Description of the invention (W) c) DXMEAQE; and d) QLDHPLPTVHPQVTYAYM (K) The single-letter symbols of the amino acids mentioned will be identified (the definitions can also be found in the text of Biochemistry, Third Editi, WH Preeaan and Conpany, c 1988 by Lubert Stryer, etc., page 21). Those skilled in the art also understand that in the sequence c), X indicates the amino acid 1 identified in the sequence and the bracket in the sequence d) indicates that the amino acid at this position may be an amino acid. As mentioned above, the present invention includes segments or truncated forms of the mentioned proteins. These proteins have one or more of the four sequences listed above as part or all of their amino acid sequences, for the purpose of patent application. In terms of the following, it means that a protein including one or more "lactam sequences" can be understood as any protein containing one of the above sequences, whether or not the protein is composed of one or more sequences a) -d) or Whether any part of the protein is formed for one or more of the above sequences. This range includes all positions on the amino acid sequence of the protein, including but not limited to the protein portion that starts and terminates the amino acid chain of the protein. It is understandable. Part of the limbic acid sequence was compared with each sequence on the Genbank database. The sequence number L 34075 is the same (Brown et al., Nature 369,756-758 (1994)). The cDHA of the SEP gene was cloned as follows: 2 mg Molt 4 cDNA (Clontech, Palo Alto, CA) IxPCR buffer (10mM TrMs-HCl, pH 8.3. 50bM KCl, lnM MgCl, 200wM dATP, 200w M dTTP, 200wM dCTP, 200bM -3 0 _ ---------- -: --- --- Packing -------- ^ Order ------ line (please read the note $ on the back before filling this page) This paper standard applies to the H® house standard rate (CNS > A4 Grid (210X297 mm) 404951 A7 B7 Printed by the Central Government Bureau of the Ministry of Economic Affairs, Industrial and Consumer Cooperatives. 5. Description of the Invention (M) dGTP; Perkin Elnex) and 1 unit of Taq Polymer Permer (Perkin Elmer). Enlargement and enrichment, 30 seconds at 94C, 30 cycles at 66 υ, 4 minutes at 66 υ, 72t: 10 minutes at the bottom, and three separate reactions with one of the following oligomer pairs: CGATCGGTCGACTGCAGCACnTGGGGATTATTGCTCTC and GCGGCCGCAGCTTTCTTCATGCATGACAACAGCCCAGGC; GCGGCCGCAAGCrTCAAGTATGCAACCiCTCTGCGGCAAGA and CCATCGGTCGACACCTTCTGCATCAGAGTCAAGTGGTCA; ^: 'GCGGCCGCAAGCrrCCTCAGCTCACATCCTTAGAGCTGCA and CGATCGGTCGACTTATTACCAGAAA sigma and sigma sigma sigma and sigma sigma sigma and sigma sigma Gassin and Anne Lang. Verlag Chenie, LFA, 1982). Do not name the PCR products of SEP3, SEP4 and SEP5 in a buffer containing T4DHA ligase (1 unit) and a 50ng pcII (TA selection kit, Invitrogen. San Diego, CA) containing a PCR product that can be modified to actively bind PCR products. Incubate at 15t: overnight to produce PCR-pcII ligated products. The PCR-pcII product was transformed into competent E.coli INVaF cells commercially available from Invitrogen. Miniprep DKA was selected from Quiagen niniprep kits (Quiagen, Chatsvorth, CA) and selected size colonies containing PCR products of appropriate size were digested with commercially available Hindlll or Sail to limit digestion, electrophoresis, and comparison with standard samples to confirm. Sep2 and Sepl cDNA were first synthesized using Tine Saver eDNA synthesis kit (Pharnacia, Piscataway, NJ) and CGATCGGTCGACCAGATGAGCACATCATAGCGCTGATGA or CGATCGGTCGACAAATTCAAAGCTGCCAAGCGTTCGGAG ·. Sep2 and Sep 1 use TineSaver cDNA synthesis kit to add 250pmoles of GCGGCCGCAAGCTTTGQCTCGAGCAATGGGGCCAGGCA?!? GCGGCCGCAAGCTTAAGATGCTTGGAACCGCACCTGCCG, perform e -31---------- install---(read W Read the notes on the reverse side and fill in this page again.) The printed paper is in accordance with the national standard of the country (CNS > A4 size (210X297mm) 404951. Printed by the Central Government Bureau of the Ministry of Economic Affairs, Shelley Consumer Cooperatives. «A7 B7 V. Description of the invention ( >) Sep 2 and Sepl cDHA thereafter were used CGATCGGTCGACCAGATGAGCACATCATAGCGCTGATGA and GCGGCCGCAAGCnTGGCTCGAGCAATGGGGCCAGGCA or GCGGCCGCAAGCITAAGATGCTTGGAACCGCACCTGCCG and CGATCGGTCGACAAATTCAAAGCTGCCAAGCGTTCGGAG, be enlarged .SeP2 PCR product described above of PCR using ΤΑ cloning kit (Invitrogen then subcloned into pell in .Sei > I PCR product is Cut with Hindlll and Sail, and separate it from the pcII vector by agarose electrophoresis. SepI (HindIII-SalI) K segment was isolated using Pharaacia's Sephasels bandprep kit and described (Saebrook et al., Molecular * Cloning Cold Spring Har * bor »1989) breeding to pUC19 Hindlll and Sail positions. Separate Sep2 (HUdIII, AspI) and Sep3 (AspI, SalI) K segments or SeP4 (HindIII, AccIII / Mrol) and Sep5UccIII / MroI, SalI fragments to the pUC18 (111110111, 5811 > vector And using this well-known technique (53 «11 ^ 〇〇11 et al., Μ 1 ο 1 ecu 1 ar C 1 ο ning C ο 1 d S pr ng arbor 19 8 9) to perform E. coli IHVct The transformation of F cells (Invitrogen) can obtain POC18-Sep23 and pUC18-Sep45, which additionally contain the sequences 1468-5326 but nucleotides and Nucleic acid. The connection and transformation of p (JC19-Sepl (EcoRV, Sail), Sep23 (EcrRV, BstEII) and Sep45 (BstEII, SalI) H segments have the ability to intestinal bacteria IN «F cells (Invitrogen) can be a well-known technology Performed (eg Sambrook et al., Molecular Cloning Cold Spring Harbor, 1989) to obtain a full length selection. The nucleic acid sequence encoding this protein is shown in Sequence No. 1. GST-SEP (g 1 utathi ο ne S transferase-s ί r ο 1 imus -32- ------.---- install ----- ^ ---: order ----- F line {please read the notes on the back before filling in this (Page) This paper size is applicable to China's domestic standard (CNS) A4 specification (210X297 mm) 404951 Printed by the Bayer Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (w) effector .protein) Sep4 and Sep5H segments For example, in the next breeding of pGEX-KG (Guan, K. and Dixon, JE (1991) Anal. Bioche · 192, 262-267). Briefly, Sep 4 was cut with a commercially available Hindlll restriction enzyme, the cut was cut with a Klenow fragment (Gibco) of NDA polymerase, and the DNA was extracted with phenol-nitroform and ethanol-precipitated using this well-known technique (Sanbrook et al., Molecular Cloning Cold Spring Harbor 1989). Sep4 (Hind 111-K1 eηow) was cut with Mrol restriction enzyme, and separated by pcII carrier with agarose electrophoresis and isolated into M segment SEP4-HindIII- The Klenow-MroI Sep5H segment was prepared by incision with Sail and Mrol. It was separated from the PCII by agarose swimming and separated into fragments SEP5-SalI-MroI. pGEX-KG (Guan, K. and Dixon, JE (1991) Anal. Biocheio. 192, 262-267) was cut with Ncol, filled with the KlenowK segment of DNA polymerase, and DHA was extracted with phenol-aerodynamics and this was used. Alcohol precipitation is performed by well-known techniques (Sanbrook et al., M e 1 ecu 1 ar C 1 o ning C o 1 d S pring H arbor, 189 9). pGEX-KG (HcoI, Klenow) was cut with Sail, and the unnamed carrier was separated by agarose electrophoresis and isolated into PGEX-KG-Ncol-Klena * -Sail, using techniques well known in the art. Since the sequence and linker of GST are the same as previously reported (Guan, K. and Dixon, JE (1991) Anal. Biochen. 192, 262-267), the protein sequence of this fusion protein is started from the sequence Ko.2 Linker between GST and SEP 45. The flag sequence can use an oligo nucleic acid including the coding sequence Asp Tyr Lys Asp Asp Asp Asp Lys, added to the carboxyl terminus of SEP or SEP, SEP4, 5 or other fragments · 33- ----------- Installation — «__ (Please read the precautions on the back before filling this page). 丨 The size of the paper is applicable to the Chinese National Standard (CNS) Α4 specification (210 ×: 297) (%) 404951 A7 B7 printed by Shellfish Consumer Cooperative of Central Government Bureau of the Ministry of Economic Affairs. 5. Description of invention (ο). Conjugated proteins can be isolated by affinity chromatography using anti-flag specific antibodies using IBI, He »Haven, Conn. Kits. Contains the aforementioned pUC19-Sepl (nucleotide 1785 in Sequence 1), PUC18-Sep23 (nucleotide 1468-5326 in Sequence 1), pUC18-Sep45 (nucleotide 4964-7 653 in Sequence 1) or? (^ ¥-$ 6 {> 45 (SEQ ID NO. 2) the transformed host cells of plastids were deposited at the Institute of Food Industry Development Center on March 7, 1995, and their registration numbers were 940075, 940076, 94 00 74 and 940073. Transformed host containing pUC19-Sepl (EcoRV, SalI) Sei > 23 (£ <; 00,831 £ 11) and 545 (& 31 £ 11,5811) fragments shown in sequence 1 The cells have been deposited in ATCC, 1 2 3 0 1P ar * k 1 awn D rive, R 0ckvi 1 1 e, May land 20852, US A, and their deposit numbers are 69756, 69753, 69754 and 69755, Fen Shi Example 3 The 210 kDa protein of the present invention was isolated using the following rapamycin analogs using the techniques described in the examples: a) 42-deoxy-42- [1- (1,1-dimethylethylamino) ) -2-GasylethylGasyl] Rapamin (described in US Patent No. 5,233,036); b) 42- [0-[(l,: l-dimethylethyl) dimethylformamide Silyl]] rapamycin (described in U.S. Patent No. 5,120,842); c) dimethylethoxy) carbonyl] -N-methylglycine rapamycin 42-ester (described in US Patent No. 5,130,307); d) with 5- (1, 1-dimethylethylamino) -2-[[((1,1-dimethylethylamino) carbonyl] limyl] -5-aminovaleric acid ethyl acetate solvate 3/4 hydrate -34- (Please read the note on the back of the page before filling in this page) This paper size is applicable to the Chinese National Standard (CNS) Λ4 Washing (210X297 mm) The Central Standards Bureau of the Ministry of Economic Affairs, Shellfish Consumer Cooperation Du printed 404351 V. Description of the invention (^) Miner's ash emblem 42-ester (refer to US Patent No. 5,130,307); e) N- [U, budimethylethoxy) glassyl] glycinaldehyde Glycerin Hydrate Rapapin 42-08 (refer to U.S. Patent No. 5,130.307 >; and f) Η2 · 燹 6-1 [(1, dimethyldimethylethoxy) kisyl]- L-lysine rapamycin 42-vinegar (refer to H patent No. 5,130,307). Serial Hao Shen Table (1) General Information (i) Applicant: Μ ο 1 π ar-K iaber, K ather ί π e L. Failli, Amedeo F. Caggiano, Thomas J. Nakanishi.Koji Chen.Yanqiu (ii) Name of the invention * · Effector protein of Rabelin (iii) Number of sequences: 2 (iv) Contact address: (A) Address: RonaId W · Alice, A aerican Home Products Corporation (B) Street name: 5 G ira 1 da F ar brews (C) City name: Madison (D) State name: New Jersey (E) Country: United States (F) Zip code: 07940-0874 -35- ---------- ; --- r-install ----: ----: order = ------ line (please read the precautions on the back before filling this page) This paper size applies to Chinese National Standard (CNS) A4 Specification (21〇 × 297 mm) 404Θ51 A7 B7 V. Description of the invention (from) Staff Cooperative Association of the Central Standards Bureau of the Ministry of Economic Affairs (V) Computer available form: (A) Media form: chip, 3.5in, 1.4 Hb capacity (B) Computer: Apple Macintosh (C) Operating system: Macintosh 7.1 (D) Software: Microsoft text (vi) Current application information: (A) Application number: (B) Date of filing: (C) Classification: (vii) Previous Please provide information: (A) Application number: U8 08 / 312,023 (B) Date of filing: September 26, 994 (C) Application number: US 0 8/20 7, 9 7 5 (D) Date of filing: March 8, 1994 (viii > Lawyer / Agent Information: (A) Name ".: Eck.Steren R · (B > Registration Number: 36, 126 (C) Reference / Record Number: AHP-93 1 67- 2-C2 (ix) Telecommunication information: (A) Telephone: (6 1 0) 90 2 -2 628 (B) Fax: (610) 688- 2 0 7 3 (2) Information of serial ID number 1: ( i) Special features of the sequence: (A) Spread: Amino acid 36 This paper is applicable to the Chinese National Standard (European People) Rule 4 is {210 > < 297mm) Please read Note I of it first;订 ^ 04951 A7B7 V. Description of the invention (v) (B) Type: Nucleic acid (C) Amine: Meaning of double-stranded DNA strand (D) Topology: Straight line (ii) Molecular type: Double-stranded cDNA to bRNA Direction of meaning (iii) Hypothesis: None (iv) Meaning: None (vi) Origin: Organ: molting 4 Human T-cell leukemia cell (B) strain: ATCCCRL 1 5 8 2 i 2 i (xi) Sequence description: Serial ID number: 1 Please Η it first .. Note sheet AAG ATG CTT GGA A CC GGA CCT Leu Gly

Met 1 AAT GTG Asn Val

Thr Gly Pro 5 GCC GCC 2Ί Ala Ala GCC ACC ACC GCT Thr Ala AGC GTC CTG CAG CAG Leu Gin Gin

Ser Val 20 ACC AGG GCC AAA GCC GCC AAG Thr Arg Ala I »ys Ala Ala Lys 40 GAG ATG AGT CAA GAG GAG TCT Ser Gin TTT GCC 81 Phe Ala 25 GAG CTC 135 Glu Leu

Ala Thr 10 AGT GGC Ser Gly GCC ACC Ala Thr CTA AAG AGC CGG Leu Lys Ser Arg 30

CAG CAC TAT GTC ACC ATG Tyr VaX

Gin His 45 Staff Cooperative Cooperative Seal of the Central Standards Bureau of the Ministry of Economic Affairs «.

Glu MET 55 TTT GAA TTG GTT Phe Glu Leu Val 75

Glu Glu Ser 60 TCC AGC TCA Ser Ser Ser ACT CGC 189 Thr Arg TTC TAT Phe Tyr GAC CAA Asp Gin 65

Thr MET 50 CTG AAC Leu Asn ACA TCT AGC 54 Thr Ser Ser 15 AAT GAG GAA 108 Asn Glu Glu 35 GAA CTC CGA 162 Glu Leu Arg Order ATA GCTIle Ala 90 AGC CTC ATA GGA GTGSer Leu lie Gly Val 95 GAT GCC AAT GAG 243 Asp Ala Asn Glu 80 GAA GGT GGG AAT 297Glu Gly Gly Asn 100 AGG AAA GGT GGC Arg Lys GCC ACCAla Thr

Gly Gly 85 CGA ATTArg lie CAT CAC ATT 216 His His lie 70 ATC TTG GCC 270 lie Leu Ala GGC AGA TTT 324 Gly Arg Phe 105 -37 This paper uses China Gujia Mou (CNS) A4 specifications &210; 2. 97 mm) 404951 A7 B7 V. Description of the invention (ice GCC AAC Ala Asn TAT CTT CGG AAC CTC CTC Arg Asn

Tyr Leu 110

Leu Leu 115 GCA TCC AAG GCC ATT GGC CGT CTT Ala Ser Lys Ala lie Gly Arg Leu 130 TAC GTG GAA TTT Tyr Val Glu Phe 145 GAG GTG AAG CGA Glu Val Lys Arg 150 CCC TCC AAT GAC CCA GTT GTC ATG GAA ATG 351 378 Pro Ser Asn Asp Pro Val Val MET Glu MET 120 \? Λ GCC ATG GCA GGG GAC ACT TTT ACC GCT GAG 405 432 Ala MET Ala Gly Asp Thr Phe Thr Ala Glu 135 140 GCC CTG GAA TGG CTG GGT GCT.GAC CGC AAT 459 486 Ala Leu Glu Trp Leu Gly Ala Asp Arg Asn 155 160 GAG GGC CGG AGA CAT GCA Glu Gly Arg Arg His Ala 165 GCT GTC CTG GTT CTC CGT GAG CTG GCC ATC AGC GTC 513 540 Ala Val Leu Val Leu Arg Glu Leu Ala He Ser Val 170 175 CCT ACC TTC TTC TTC CAG Phe Phe

Pro Thr 180

Phe Gin 185 CAA GTG CAA CCC TTC TTT GAC AAC ATT TTT GTG GCC 567 594 Gin Val Gin Pro Phe Phe Asp Asn lie Phe Val Ala. 190 195 GTG TGG GAC CCC Val Trp Asp Pro 200 AAA CAG GCC ATC Lys Gin Ala lie 205 CGT GAG GGA GCT GTA GCC GCC CTT CGT GCC 621 646 Arg Glu Gly Ala Val Λίβ Ala Leu Arg Ala 210 215 TGT CTG Cys Leu ATT CTC ACA ACC lie Leu Thr Thr 220 CAG CGT GAG CCG AAG GAG ATG CAG AAG CCT CAG TGG 702 Gin Arg Glu Pro Lys Glu MET Gin Lys Pro Gin Trp 225 230 Printed by TAC AGG CAC ACA Tyr Arg His Thr 235 GAG AAG GGC ATG Glu Lys Gly MET 255 TTT GAA GAA GCA Phe Glu Glu Ala 240 GAG AAG GGA 729 Glu Lys Gly TTT GAT GAG ACC TTG GCC AAA 756 Phe Asp Glu Thr Leu Ala Lys 245 250 AAT CGG GAT GAT CGG ATC CAT GGA GCC TTG TTG ATC CTT AAC 783 810 Asn Arg Asp Asp Arg lie His Gly Ala Leu Leu lie JLeu Asn 260 265 GAG CTG GTC CGA ATC AGC Val Arg

Glu Leu 270 GAA ATC ACA CAG Glu He Thr Gin 290

He Ser 275CAG CAG Gin Gin AGC ATG GAG GGA GAG CGT CTG AGA GAA GAA ATG GAA 837 864 Ser MET Glu Gly Glu Arg Leu Arg Glu Glu MET Glu 280 285

Leu Val 295 GTA CAC GAC AAG TAC 891 His Asp Lys

GAT CTC ATG 918

Tyr Cys Lys Asp Leu i4t.T Giy 300 305 -38- The standard of this paper is not applicable to the National Solid Standard (CNS) A4 specification (210X297 mm) 404951 A7 B7 V. Description of the invention (47 TTC GGA ACA AAA CCT CGT CAC Phe Gly Thr Lys Pro Arg His 310 CCC CAG CAG TCA AAT GCC TTG Gin Ser

Pro Gin 325 CTC ATG Leu MET

Asn Ala Leu 330 ATT ACC CCC '945 lie Thr Pro 315 GTG GGG CTG 999 Val Gly Leu TTC ACC Phe Thr AGT TTC Ser Phe GGA TTT GGG ACC TCC Gly Thr Ser

Gly Phe 345 CGG TGT TGC AGA GAC TTG ATG Cys Arg

Arg Cys 360 CTG AAA TGC AGG Leu Lys

Cys Arg 380

Asp Leu MET 365 AAT AGC AAG Asn Ser Lys CCC AGT CCA 1053 Pro Ser Pro 350 GAG GAG AAA 1107 Glu Glu Lys CTG GGG TAC AGC Leu Gly Tyr Ser 335 GCT AAG TCC ACC Ala Lys Ser Thr 355 CAG GCT GTA CAG 972 Gin Ala Val Gin 320 TCT CAC CA / V Gnc 1026 Ser His Gin Gly 340 CTG GTG GAG AGC 1080 Leu Val Glu Ser TTG CCC Leu Pro CAA GAT Gin Asp 415 GCG GCC Ala Ala CGC TTG GCT GCA TTC Arg Leu Ala Ala Phe 400 ACC ATG AAC CAT GCC Thr MET Asn His Ala 420 AAC TCG CTG 1161 Asn Ser Leu 385 CGA CCT TCT 1215 Arg Pro Ser 405 CTA AGC TGT 1269 Leu Ser Cys TTT GAT CAG GTG TGC CAG Phe Asp Gin Val Cys Gin 370 375 ATC CAA ATG ACA ATC CTT He Gin MET Thr lie Leu 390 GCC TTC Ala Phe ACA GAT ACC CAG Thr Asp GTC AAG AAG GAG Lys Glu TGG GTG 1134 Trp Val (please read the “Precautions on the back page before this page”) TTC CAA Phe Gin 435 GCC CTG GGG Ala Leu Gly Employees of the Central Standards Bureau of the Ministry of Economic Affairs Lambda Cooperative printed GTC TAT TTG CCT CGC GTG CTG Leu Pro

Val Tyr 450 TTC GCC CAT AAG Phe Ala

His Lys 470 ATC AGC lie Ser

Arg Val Leu 455 AGG CAG AAG Arg Gin Lys CTA CTT TCT 1323 Leu Leu Ser. 440 GAC ATC ATC 1377 Asp lie lie

Val Lys 425 GTG GCT Val Ala

Thr Gin 410 AAG GAA Lys Glu AAT TTG 1188 Asn Leu 395 TAT CTC 1242 Tyr Leu GTG AGG TCT GAG Val Arg Ser Glu 445 CGT ACA 1296 Arg Thr 430 TTT AAG 1350 Phe Lys ATG CTG GCT CGA GCA MET Leu Ala Arg Ala 490 ATG CAG 1431 Ala MET Gin 475 ATG GGG CCA 1485 MET Gly Pro 495 CGA GCG GCC CTG CCC CCA Arg Ala Ala Leu Pro Pro 460 465 GTG GAC GCC ACA GTC TTC Val Asp Ala Thr Val Phe 480 GGC ATC Gly lie CAG CAG GAT ATC Gin Gin Asp lie 500 AAG GAC 1404 Lys Asp ACT TGC 1458 Thr Cys 485 AAG GAG 1512 Lys Glu This paper arc ruler agricultural suitable Λ middle β national standard (GNS > Α4 size (210X297 mm) 404951 A7 B7 V. Description of the invention (4) CTG CTG GAG CCC ATG CTG GCA GTG GGA CTA AGC 1539 Leu Leu Glu Pro MET Leu Ala Val Gly Leu Ser 505 510 Member of the Central Standards Bureau of the Ministry of Economic Affairs'- :: Cooperative printed TAC GAC CTG AGC CGT CAG ATT CCA CAG CTA AAG 1593 Tyr Asp Leu Ser Arg Gin lie Pro Gin Leu Lys 525 5.30 CTG AAA ATG CTG TCC CTG GTC CTT ATG CAC AAA 1647 Leu Lys MET Leu Ser Leu Val Leu MET His Lys 540 545 550 CCC AAG GGC CTG GCC CAT CAG CTG GCC TCT CCT 1701 Pro Lys Gly Leu Ala His Gin Leu Ala Ser Pro 560 565 GCC AGC GAT GTG GGC AGC ATC ACT CTT GCC CTC 1755 Ala Ser Asp Val Gly Ser lie Thr Leu Ala Leu 580 585 TTT GAA GGC CAC TCT CTG ACC CAA TTT GTT CGC 1809 Phe Glu Gly His Ser Leu Thr Gin Phe Val Arg 595 600 AAC AGT GAG CAC AAG GAG ATC CGC ATG GAG GCT 1863 Asn Ser Glu His Lys Glu lie Arg MET Glu Ala 615 620 CTC ACA CCC TCC ATC CAC CTC ATC AGT GGC CAT 1917 Leu Thr Pro Ser lie His Leu lie Ser Gly His 630 635 640 GCA GTG CAA GTG GTG GCA GAT GTG CTT AGC AAA 1971 Ala Val Gin Val Val Ala Asp Val Leu Ser Lys 650 655 GAT CCT GAC CCT GAC ATT CGC TAC TGT GTC TTG 2025 Asp Pro Asp Pro Asp lie Arg Tyr Cys Val Leu 670 675 GAT GCA CAC CTG GCC CAG GCG GAG AAC TTG CAG 2079 Asp Ala His Leu Ala Gin Ala Glu Asn Leu Gin 685 " 690 CCT GCC CTC ACT GCA GTG CTC 1566 Pro Ala Leu Thr Ala Val Leu 515 520 AAG GAC ATT ΟΑΛ GAT GGC, ΓΤΛ 1620 Lys Asp lie Gin Asp Gly Leu 535 CCC CTT CGC CAC CCA GGC ATG 1674 Pro Leu Arg His Pro Gly MET 555 GGC CTC AC G ACC CTC CCT GAG 1728 Gly tea Thr Thr Leu Pro Glu 570 575 CGA ACG CTT GGC AGC TTT GAA 1782 Arg Thr Leu Gly Ser Phe Glu 590 CAC TGT GCG GAT CAT TTC CTG 1836 His Cys Ala Asp His Phe Leu 605 610 GCC CGC ACC TGC TCC CGC CTG 1890 Ala Arg Thr Cys Ser Arg Leu 625 GCT CAT GTG GTT AGC CAG ACC 1944 Ala His Val Val Ser Gin Thr 645 CTG CTC GTA GTT GGG ATA ACA 1998 Leu Leu Val Val Gly lie Thr 660 665 GCG TCC CTG GAC GAG CGC TTT 2052 Ala Ser Leu Asp Glu Arg Phe 680 GCC TTG TTT GTG GCT CTG AAT 2106 Ala Leu Phe Val Ala Leu Asn 695 700 Please make the first page of this page. The paper size is applicable to 0 Chinese standards (CNS) Α4 specifications U10X297 mm)

U ^ 04951 V. Description of the Invention (β) A7 B7 GAC CAG GTG TTT GAG ATC Asp Gin Val Phe Glu lie 705 AGC ATG AAC CCT GCC TTT Asn Pro

Ser MET 720 TTG ACA GAG TTG Leu Thr

Glu Leu 740

Ala Phe 725 GAG CAC Glu His CGG GAG CTG 2133 Arg Glu Leu 710 GTC ATG CCT 2187 Val MET Pro GCC ATC TGC ACT GTG Ala lie Cys Thr Val 715 GGC CGA CTC AGT 2160 Gly Arg Leu Ser

TTC CTG CGC AAG ATG CTC ATC Lys MET ATG CTG MET Leu GGG CAC CTG GTC Gly His CCT ATT CTG AAG Pro lie Leu Lys 775 CCA GGT GTG ATC Pro Gly Val lie 795 GGC CTG Gly Leu 810

GAA ATG Glu MET CTC CAG GAT TCC Leu Gin

Asp Ser 830

Leu Val 760 GCA TTA Ala Leu AAT AAT Asn Asn AGG ΑΑΆ Arg Lys 815 TCT TTG Ser Leu AGT GGG ATT 2241 Ser Gly lie 745 TCC AAT GCC 2295 Ser Asn Ala ATT TTG AAA 2349 lie Leu Lys 780 GTC CTG GCA 2403 Val Leula 800 TGG GTT GAT 2457 Trp Val Asp

Phe Leu Arg 730 GGA AGA ATC Gly Arg lie

Lys Glu 75a

Leu lie 735 CAG ATT 2214 Gin lie GAG CAG AGT Gin Ser CCC CGA CTC ATC CGC CCC TAG lie Arg

Pro Arg Leu 765

CTG

Leu Lys Asp 785 ACA ATA GGA Thr He Gly GAT CCA GAC Pro Asp GAA TTG Glu Leu 805

Pro Tyr 770 CCT GAT Pro Asp GCA CAG Ala Gin GCC CGC 2268 Ala Arg 755 ATG GAG 2322 MET Glu CCA AAC '2376 Pro Asn 790 GTT AGT 2430 Val Ser : Cooperative printed CAG TTG Gin Leu GTG GCC AGC ACT Val Ala Ser Thr 850 TTG GCC AAA 2511 Leu Ala · Lys 835 GGC TAT GTA 2565 Gly Tyr Val TTG CTT GAG GTG CTA CTG Leu Leu Glu Val Leu Leu 865 AGA GAG GCC ATC CGT GTG Arg Glu Ala He Arg Val 885 AAT TTT CTG 2619 Asn Phe Leu 870 TTA GGG CTT 2673 Leu Gly Leu 890 GAA CTT TTT ATT ATC Glu Leu Phe lie lie 820 AGG CAG GTG GCT CTG Arg Gin Val Ala Leu 840 GTA GAG CG TAC AGG Val Glu Pro Tyr Arg 855 AAG ACT GAG CAG AAC Lys Thr Glu Gin Asn 875 TTA GGG GCT Leu Gly Ala TTG GAT Leu Asp 895 ATC ATG GAC ATG 2484 lie MET Asp MET 825 TGG ACC CTG GGA 2538 Trp Thr Leu Gly AAG TAC CCT ACT 2592 Lys Tyr Pro Thr 860 CAG GGT ACA CGC 2646 Gin Gly Thr Arg 880 CCT TAC AAG CAC 2700 Pro Tyr Lys His -41- This paper uses China • Jiaxiong Series_ 《CNS》 A4 * L Grid (210X297 PCT) A7 B7 _ ^ 04951 V. invention is described)

AAA GTG AAC ATT GGC ATG ΑΤΑ GAC CAG TCC CGG GAT GCC TCT GCT GTC AGO CTG 2727 2754

Lys Val Asn lie Gly MET lie Asp Gin Ser Arg Asp Ala Ser Ala Val Ser Leu 900 905 910 915 TCA GAA TCC AAG TCA AGT CAG GAT TCC TCT GAC TAT AGO ACT AGT GAA ΛΤΟ CTC? 2781 2808

Ser Glu Ser Lys Ser Ser Gin Asp Ser Ser Asp Tyr Ser Thr Ser Glu MET Leu 920 925 930 935

GTC AAC ATG GGA AAC TTG CCT CTG GAT GAG TTC TAC CCA GCT GTG TCC ATG GTG 2835 2862

Val Asn MET Gly Asn Leu Pro Leu Asp Glu Phe Tyr Pro Ala Val Ser MET Val 940 945 950 •

GCC CTG ATG CGG ATC TTC CGA GAC CAG TCA CTC TCT CAT CAT CAC ACC ATG GTT 2889 2916

Ala Leu MET Arg He Phe Arg Asp Gin Ser Leu Ser His His His Thr MET Val 955 960 965_ 970

GTC CAG GCC ATC ACC TTC ATC TTC AAG TCC CTG GGA CTC AAA TGT GTG CAG TTC 2943 2970

Val Gin Ala lie Thr Phe He Phe Lys Ser Leu Gly Leu Lys Cys Val Gin Phe 975 980 985

CTG CCC CAG GTC ATG CCC ACG TTC CTT AAT GTC ATT CGA GTC TGT GAT GGG GCC 2997 3024

Leu Pro Gin Val MET Pro Thr Phe. Leu Asn Val lie Arg Val Cys Asp Gly Ala 990 995 1000 1005

ATC CGG GAA TTT TTG TTC CAG CAG CTG GGA ATG TTG GTG TCC TTT GTG AAG AGC 3051 3078 lie Arg Glu Phe Leu Phe Gin Gin Leu Gly MET Leu Val Ser Phe Val Lys Ser 1010 1015 1020 1025

CAC ATC AGA CCT TAT ATG GAT GAA ATA GTC ACC CTC ATG AGA GAA TTC TGG GTC 3105 3132

His He Arg Pro Tyr MET Asp Glu lie Val Thr Leu MET Arg Glu Phe Trp Val 1030 1035 1040

ATG AAC ACC TCA ATT CAG AGC ACG ATC ATT CTT CTC ATT GAG CAA ATT GTG GTA 3159 3186 MET Asn Thr Ser lie Gin Ser Thr lie He Leu Leu lie Glu Gin He Val Val 1045 1050 1055 1060

GCT CTT GGG GGT GAA TTT AAG CTC TAC CTG CCC CAG CTG ATC CCA CAC ATG CTG 3213 3240

Ala Leu Gly Gly Glu Phe Lys Leu Tyr Leu Pro Gin Leu lie Pro His MET Leu 1065 1070 1075

CGT GTC TTC ATG CAT GAC AAC AGC CCA GGC CGC ATT GTC TCT ATC AAG TTA CTG 3267 3294

Arg Val Pha MET His Asp Asn Ser Pro Gly Arg lie Val Set lie Lys Leu Leu 1080 1085 1090 1095 42 This paper size is applicable to China National Standard (CNS) A4 (210X297 mm) ----.---- -^ ------ ir ------ ^. ». L · ::, ¾ (please read the precautions on the back ^^ this page) Member of the Central Government Prospective Bureau of the Ministry of Economic Affairs A7 B7 404951 V. Description of the invention (4 · *)

GCT GCA ATC CAG CTG TTT GGC GCC AAC CTG GAT GAC TAC CTG CAT TTA CTG CTG 3321 3348 (Please read the notes on the back first 'this page)

Ala Ala lie Gin Leu Phe Gly Ala Asn Leu Asp Asp Tyr I »eu His Leu Leu \ Leu 1100 1105 1110/1115 CCT CCT ATT GTT AAG TTG TTT GAT GCC CCT GAA GCT CCA CTG CCA TCT CGA ΛΛΟ 3375 3402

Pro Pro lie Val Lys Leu Phe Asp Ala Pro Glu Ala Pro Leu Pro Ser Arg Lys 1120 1125 1130

GCA GCG CTA GAG ACT GTG GAC CGC CTG ACG GAG TCC CTG GAT TTC ACT GAC TAT 3429 3456

Ala Ala Leu Glu Thr Val Asp Arg Leu Thr Glu Ser Leu Asp Phe Thr Asp Tyr 1135 1140 1145 1150

GCC TCC CGG ATC ATT CAC CCT ATT GTT CGA ACA CTG GAC CAG AGC CCA GAA CTG 3483 3510

Ala Ser Arg lie lie His Pro lie Val Arg Thr Leu Asp Gin Ser Pro Glu Leu 1155 1160 1165

CGC TCC ACA GCC ATG GAC ACG CTG TCT TCA CTT GTT TTT CAG CTG GGG AAG AAG 3537 3564

Arg Ser Thr Ala MET Asp Thr Leu Ser Ser Leu Val Phe Gin Leu Gly " Lys Lys 1170 1175 1180 1185

TAC CAA ATT TTC ATT CCA ATG GTG AAT AAA GTT CTG GTG CGA CAC CGA ATC AAT 3591 3618

Tyr Gin lie Phe He Pro MET Val Asn Lys Val Leu Val Arg His Arg lie Asn 1190 1195 1200 1205

CAT CAG CGC TAT GAT GTG CTC ATC TGC-AGA ATT GTC AAG GGA TAC ACA CTT GCT 3645 3672

His Gin Arg Tyr Asp Val Leu He Cys Arg lie * Val Lys Gly Tyr Thr Leu Ala 1210 1215 1220

GAT GAA GAG GAG GAT CCT TTG ATT TAC CAG CAT CGG ATG CTT AGG AGT GGC CAA 3699 3726

Asp Glu Glu Glu Asp Pro Leu He Tyr Gin His Arg MET Leu Arg Ser Gly Gin 1225 1230 1235 1240

GGG GAT GCA TTG GCT AGT GGA CCA GTG GAA ACA GGA CCC ATG AAG AAA CTG CAC 3753 3780 Member of the Central Standards Bureau of the Ministry of Economic Affairs. Printed by Cooperative.

Gly Asp Ala Leu Ala Ser Gly Pro Val Glu Thr Gly Pro MET Lys Lys Leu His 1245 1250 1255

GTC AGC ACC ATC AAC CTC CAA AAG GCC TGG GGC GCT GCC AGG AGG GTC TCC AAA 3807 3834

Val Ser Thr lie Asn Leu Gin Lys Ala Trp Gly Ala Ala Arg Arg Val Ser Lys 1260 1265 1270 1275

GAT GAC

GAA \ CTG AGA CGG CTG AGC CTG GAG CTG CTG AAG GAC TCA 3861 3888

Asp Asp Trp Leu Glu Trp Leu Arg Arg Leu Ser Leu Glu Leu Leu Lys Asp Ser 128〇1285 1290 1295 -43 · This paper size is free to use the Chinese national rubbing < CNS > person 4 washing grid "2 丨 0X297 mm ) A7 B7 4〇495l V. Description of the invention (〇)

TCA TCG CCC TCC CTG CGC TCC TGC TGG GCC CTG GCA CAG GCC TAC AAC CCG ATG 3915 3942 (Please read the notes on the back first and fill in this page)

Ser Ser Pro Ser Leu Arg Ser Cys Trp Ala Leu Ala Gin Ala Tyr Asn Pro MET 1300 1305 1310 GCC AGG GAT CTC TTC AAT GCT GCA TTT GTG TCC TGC TGG TCT GAA CTG ΑΛΤ ΟΛΛ 3969 3996

Ala Arg Asp Leu Phe Asn Ala Ala Phe Val Ser Cys Trp Ser GIu Leu Asn Glu 1315 1320 1325 1330

GAT CAA CAG GAT GAG CTC ATC AGA AGC ATC GAG TTG GCC CTC ACC TCA CAA GAC 4023 4050

Asp Gin Gin Asp Glu Leu lie Arg Ser lie Glu Leu Ala Leu Thr Ser Gin Asp 1335 1340 1345

ATC GCT GAA GTC ACA CAG ACC CTC TTA AAC TTG GCT GAA TTC ATG GAA CAC AGT 4077 4104 lie Ala Glu Val Thr Gin Thr Leu Leu Asn Leu Ala Glu Phe MET Glu His Ser 1350 1355 1360 1365

GAC AAG GGC CCC CTG CCA CTG AGA GAT GAC AAT GGC, AXT GTT CTG CTG GGT GAG 4131 4158

Asp Lys Gly Pro Leu Pro Leu Arg Asp Asp Asn Gly lie Val Leu Leu Gly Glu 1370 1375 1380 1385

AGA GCT GCC AAG TGC CGA GCA TAT GCC AAA GCA CTA CAC TAC AAA GAA CTG GAG 4185 4212

Arg Ala Ala Lys Cys Arg Ala Tyr Ala Lys Ala Leu His Tyr Lys Glu Leu Glu 1390 1395 1400

TTC CAG AAA GGC CCC ACC CCT GCC ATT CTA GAA TCT CTC ATC AGO ATT AAT AAT 4239 4266

Phe Gin Lys Gly Pro Thr Pro Ala lie Leu Glu Ser Leu lie Ser lie Asn Asn 1405 1410 1415 1420

AAG CTA CAG CAG CCG GAG GCA GCG GCC GGA GTG TTA GAA TAT GCC ATG AAA CAC 4293 4320

Lys Leu Gin Gin Pro Glu Ala Ala Ala Gly Val Leu Glu Tyr Ala MET Lys His 1425 1430 1435

TTT GGA GAG CTG GAG ATC CAG GCT ACC TGG TAT GAG AAA CTG CAC GAG TGG GAG 4347 4374 Member of the Central Standards Bureau of the Ministry of Economic Affairs \ -Printed by the cooperative

Phe Gly Glu Leu Glu lie Gin Ala Thr Trp Tyr Glu Lys Leu His Glu Trp Glu 1440 1445 1450 1455

GAT GCC CTT GTG GCC TAT GAC AAG AAA ATG GAC ACC AAC AAG GAC GAC CCA GAG 4401 4428

Asp Ala Leu Val Ala Tyr Asp Lys Lys MET Asp Thr Asn Lys Asp Asp Pro Glu 1460 1465 1470 1475

CTG ATG CTG GGC CGC ATG CGC TGC CTC GAG GCC TTG GGG GAA TGG GGT CAA CTC 4455 4482

Leu MET Leu Gly Arg MET Arg Cys Leu Glu Ala Leu Gly Glu Trp Gly Gin Leu 1480 1485 1490 This paper size is applicable to the Chinese national kneading car (CNS > A4 size (210X297 mm) A7 B7 404951 V. Description of the invention (M )

GTT CAC CCT CAG GTG ACC TAT GCC TAC ATG AAA AAC ATG TGG AAG AGT GCC CGC 5103 5130

Val His Pro Gin Val Thr Tyr Ala Tyr MET Lys Asn MET Trp Lys Ser Ala Arg 1695 1700 1705 AAG ATC GAT GCC TTC CAG CAC ATG CAG CAT TTT GTC CAG ACC ATG CAC5 CAA ΠΛΠ 5157 5184

Lys He Asp Ala Phe Gin His MET-Gin His Phe Val Gin Thr MET Gin Gin Gin 1710 1715 1720 1725

GCC CAG CAT GCC ATC GCT ACT GAG GAC CAG CAG CAT AAG CAG GAA CTG CAC AAG 5211 5238

Ala Gin His Ala lie Ala Thr Glu Asp Gin Gin His Lys Gin Glu Leu His Lys 1730 1735 1740 1745 •

CTC ATG GCC CGA TGC TTC CTG AAA CTT GGA GAG TGG CAG CTG AAT CTA CAG GGC 5265 5292

Leu MET Ala Arg Cys Phe Leu Lys Leu Gly Glu Trp Gin Leu Asn Leu Gin Gly 1750 1755 1760

ATC AAT GAG AGC ACA ATC CCC AAA GTG CTG CAG TAC TAC AGC GCC GCC ACA GAG 5319 5346 lie Asn Glu Ser Thr lie Pro Lys Val Leu Gin Tyr Tyr Ser Ala Ala Thr Glu 1765 1770 1775 1780

CAC GAC CGC AGC TGG TAC AAG GCC TGG CAT GCG TGG GCA GTG ATG AAC TTC GAA 5373 5400

His Asp Arg Ser Trp Tyr Lys Ala Trp His Ala Trp Ala Val MET Asn Phe Glu 1785 1790 17.95

GCT GTG CTA CAC TAC AAA CAT CAG AAC CAA GCC CGC GAT GAG AAG AAG AAA CTG 5427 5454

Ala Val Leu His Tyr Lys His Gin Asn Gin Ala Arg Asp Glu Lys Lys Lys Leu 1800 1805 1810 1815

CGT CAT GCC AGC GGG GCC AAC ATC ACC AAC GCC ACC ACT GCC GCC ACC ACG GCC 5481 5508

Arg His Ala Ser Gly Ala Asn lie Thr Asn Ala Thr Thr Ala Ala Thr Thr Ala 1820 1825 1830 1835

GCC ACT GCC ACC ACC ACT GCC AGC ACC GAG GGC AGC AAC AGT GAG AGC GAG GCC 5535 5562 Member of the Central Bureau of Assistance, Ministry of Economic Affairs > h Cooperative Press

Ala Thr Ala Thr Thr Thr Ala Ser Thr Glu Gly Ser Asn Ser Glu Ser Glu Ala 1840 1845 1850

GAG AGC ACC GAG AAC AGC CCC ACC CCA TCG CCG CTG CAG AAG AAG GTC ACT GAG 5589 5616

Glu Ser Thr Glu Asn Ser Pro Thr Pro Ser Pro Leu Gin Lys Lys Val Thr Glu 1855 I860 1865 1870

GAT CTG TCC AAA ACC CTC CTG ATG TAC ACG GTG CCT GCC GTC CAG GGC TTC TTC 5643 5670

Asp Leu Ser Lys Thr Leu Leu MET Tyr Thr Val Pro Ala Val Gin Gly Phe Phe 1875 1880 1885 -46- This paper size is applicable to Chinese solid family standards < CNS > A4 specifications (210X297 male * > A7 B7 404951 5 And invention description (4r)

CGT TCC ATC TCC TTG TCA CGA GGC AAC AAC CTC CAG GAT ACA CTC AGA GTT CTC 5697 5724

Arg Ser lie Ser Leu Ser Arg Gly Asn Asn Leu Gin Asp Thr Leu Arg Val Leu 1890 1895 1900 190Π

ACC TTA TGG TTT GAT TAT GGT CAC TGG CCA GAT GTC AAT GAG GCC TTA GTG GAG 5751 5778

Thr Leu Trp Phe Asp Tyr Gly His Trp Pro Asp Val Asn Glu Ala Leu Val Glu 1910 1915 1920 1925

GGG GTG AAA GCC ATC CAG ATT GAT ACC TGG CTA CAG GTT ATA CCT CAG CTC ATT 5805 5832

Gly Val Lys Ala lie Gin lie Asp Thr Trp Leu Gin Val lie Pro Gin Leu lie 1930 1935 1940

GCA AGA ATT GAT ACG CCC AGA CCC TTG GTG GGA CGT CTC ATT CAC CAG CTT CTC 5859 5886

Ala Arg lie Asp Thr Pro Arg Pro Leu Val Gly Arg Leu lie His Gin Leu Leu 1945 1950 1955 1960

ACA GAC ATT GGT CGG TAC CAC CCC CAG GCC CTC ATC TAC CCA CTG ACA GTG GCT 5913 5940

Thr Asp lie Gly Arg Tyr His Pro Gin Ala Leu lie Tyr Pro Leu Thr Val Ala 1965 1970 '1975

TCT AAG TCT ACC ACG ACA GCC CGG CAC AAT GCA GCC AAC AAG ATT CTG AAG AAC 5967 5994

Ser Lys Ser Thr Thr Thr Ala Arg His Asn Ala Ala Asn Lys lie Leu Lys Asn 1980 1985 1990 1995

ATG TGT GAG CAC AGC AAC ACC CTG GTC CAG CAG GCC ATG ATG GTG AGC GAG GAG 6021 6048 MET Cys Glu His Ser Asn Thr Leu Val Gin Gin Ala MET MET Val Ser Glu Glu 2000 2005 2010 2015

CTG ATC CGA GTG GCC ATC CTC TGG CAT GAG ATG TGG CAT GAA GGC CTG GAA GAG 6075 6102

Leu lie Arg Val Ala He Leu Trp His Glu MET Trp His Glu Gly Leu Glu Glu 2020 2025 2030

GCA TCT CGT TTG TAC TTT GGG GAA AGG AAC GTG AAA GGC ATG TTT GAG GTG CTG Printed by V X Cooperative, Central Standards Bureau, Ministry of Economy 6129 6156

Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly MET Phe Glu Val Leu 2035 2040 2045 2050

GAG CCC TTG CAT GCT ATG ATG GAA CGG GGC CCC CAG ACT CTG AAG GAA ACA TCC 6183 6210

Glu Pro Leu His Ala MET MET Glu Arg Gly Pro Gin Thr Leu Lys Glu Thr Ser 2055 2060 2065

TTT AAT CAG GCC TAT GGT CGA GAT TTA ATG GAG GCC CM GAG TGG TGC AGG AAG 6237 6264

Phe Asn Gin Ala Tyr Gly Arg Asp Leu MET Glu Ala Gin Glu Trp Cys Arg Lys 2070 2075 2080 2085 -47- This paper size applies to Chinese national standards (CNS > A4 grid (210X297 mm) 404951 A7 B7 V. Invention Explanation (枓) Printed by TAC ATG AAA TCA GGG AAT GTC AAG GAC CTC ACC CAA GCC TGG GAC CTC TAT TAT 6291 6318 Tyr MET Lys Ser Gly Asn Val Lys Asp Leu Thr Gin Ala Trp Asp Leu Tyr Tyr 2090 209S 2100 2105 CAT GTG TTC CGA CGA ATC TCA AAG CAG CTG CCT CAG CTC ACA TCC TTA GAG CTG 6345 6372 His Val Phe Arg Arg He Ser Lys Gin Leu Pro Gin Leu Thr Ser Leu Glu Leu 2110 2115 2120 CAA TAT GTT TCC CCA AAA CTT CTG ATG TGC CGG GAC CTT GAA TTG GOT GTG CCA 6399 6426 Gin Tyr Val Ser Pro Lys Leu Leu M £ T Cys Arg Asp Leu Glu Leu Ala Val Pro 2125 2130 2135 2140 GGA ACA TAT GAC CCC AAC CAG CCA ATC ATT CGC ATT CAG TCC ATA GCA CCG TCT 6453 6480 Gly Thr Tyr Asp Pro Asn Gin Pro lie lie Arg lie Gin Ser lie Ala Pro Ser 2145 2150 2155 TTG CAA GTC ATC ACA TCC AAG CAG AGG CCC CGG A AA TTG ACA CTT ATG GGC AGC 6507 ^ 6534 Leu Gin Val lie Thr Ser Lys Gin Arg Pro Arg Lys Leu Thr Leu MET Gly Ser 2160 2165 2170 2175 AAC GGA CAT GAG TTT GTT TTC CTT CTA AAA GGC CAT GAA GAT CTG CGC CAG GAT 6561 6588 Asn Gly His Glu Phe Val Phe Leu Leu Lys Gly His Glu Asp Leu Arg Gin Asp 2180 2185 2190 2195 GAG CGT GTG ATG CAG CTC TTC GGC CTG GTT AAC ACC CTT CTG GCC AAT GAC CCA 6615 6642 Glu Arg Val MET Gin Leu Phe Gly Leu Val Asn Thr Leu Leu Ala Asn Asp Pro 2200 2205 2210 ACA TCT CTT CGG AAA AAC CTC AGC ATC CAG AGA TAC GCT GTC ATC CCT TTA TCG 6669 6696 Thr Ser Leu Arg Lys Asn Leu Ser He Gin Arg Tyr Ala Val lie Pro Leu Ser 2215 2220 2225 2230 ACC AAC TCG GGC CTC ATT GGC TGG GTT CCC CAC TGT GAC ACA CTG CAC GCC CTC 6723 6750 Thr Asn Ser Gly Leu lie Gly Trp Val Pro His Cys Asp Thr Leu His Ala Leu 2235 2240 2245 ATC CGG GAC TAC AGG GAG AAG AAG AAG ATC CTT CTC AAC ATC GAG CAT CGC ATC 6777 6804 lie Arg Asp Tyr Arg Glu Lys Lys Lys lie Leu Leu Asn lie Glu His Arg lie 2250 2255 2260 --J2265, ATG TTG CGG ATG GCT CCG GAC TAT GAC CAC TTG ACT CTG ATG CAG AAG GTG GAS 6831 ^ 858 MET Leu Arg MET A 丄 a Pro Asp Tyr Asp Kis Leu Thr Leu MET Gin Lys Vai Giu 2270 2275 2280 2285 I, ~ <. Packing. — • Order --- --- Ball (please read the back < Note 1 ^^-then fill out this page) This paper size is applicable to the Chinese National Standard (CNS) M size (210X297 mm) A7 B7 6939 ^ 04951 V. Description of the invention (〇)

GTG TTT GAG CAT GCC GTC AAT AAT ACA GCT GGG GAC GAC CTG GCC AAG CTG CTG 6885 6912

Val Phe Glu His Ala Val Asn Asn Thr Ala Gly Asp Asp Leu Ala Lys Leu Leu 2290 2295 2300

TGG CTG AAA AGC CCC AGC TCC GAG GTG TGG TTT GAC CGA AGA ACC AAT ΤΛΤ ACC 6966

Trp Leu Lys Ser Pro Ser Ser Glu Val Trp Phe Asp Arg Arg Thr Asn Tyr Thr 2305 2310 2315 2320

CGT TCT TTA GCG GTC ATG TCA ATG GTT GGG TAT ATT TTA GGC CTG GGA GAT AGA 6993 7020

Arg Ser Leu Ala Val MET Ser MET Val Gly Tyr lie Leu Gly Leu Gly Asp Arg 2325 2330 2335

CAC CCA TCC AAC CTG ATG CTG GAC CGT CTG AGT GGG AAG ATC CTG CAC ATT GAC 7047 7074

His Pro Ser Asn Leu MET Leu Asp Arg Leu Ser Gly Lys lie Leu His lie Asp 2340 2345 2350 2355

TTT GGG GAC TGC TTT GAG GTT GCT ATG ACC CGA GAG AAG TTT CCA GAG AAG ATT 7101 7128

Phe Gly Asp Cys Phe Glu Val Ala MET Thr Arg Glu Lys Phe Pro Glu Lys He 2360 2365 2370 2375

CCA TTT AGA CTA ACA AGA ATG TTG ACC AAT GCT ATG GAG GTT ACA GGC CTG GAT 7155 7182

Pro Phe Arg Leu Thr Arg MET Leu Thr Asn Ala MET Glu Val Thr Gly Leu Asp 2380 2385 2390

GGC AAC TAC AGA ATC ACA TGC CAC ACA GTG ATG GAG GTG CTG CGA GAG CAC AAG 7209 7236

Gly Asn Tyr Arg lie Thr Cys His Thr Val MET Glu Val Leu Arg Glu His Lys 2395 2400 2405 2410

GAC AGT GTC ATG GCC GTG CTG GAA GCC TTT GTC TAT GAC CCC TTG CTG AAC TGG 7263 7290

Asp Ser Val MET Ala Val Leu Glu Ala Phe Val Tyr Asp Pro Leu Leu Asn Trp 2415 2420 2425

AGG CTG ATG GAC ACA AAT ACC AAA GGC AAC AAG CGA TCC CGA ACG AGG ACG GAT 7317 7344

Arg Leu MET Asp Thr Asn Thr Lys Gly Asn Lys Arg Ser Arg Thr Arg Thr Asp 2430 2435 2440 2445

TCC TAC TCT GCT GGC CAG TCA GTC GAA ATT TTG GAC GGT GTG GAA CTT GGA GAG 7371 * 7398

Ser Tyr Ser Ala Gly Gin Ser Val Glu He Leu Asp Gly Val Glu Leu Gly Glu 2450 2455 2460 2465

CCA GCC CAT AAG AAA ACG GGG ACC ACA GTG CCA GAA TCT ATT CAT TCT TTC ATT 7425. 7452

Pro Ala His Lys Lys Lys Thr Glv Thr Thr Val Pro Glu Ser lie His Ser Phe He 2470. 2 ^ fb 2480 -49- This paper size is for Chinese National Standard (CNS) A4 (210X297 mm), please first a Face note $ Item page Printed by the Central Standards Bureau of the Ministry of Economic Affairs V. 乂 D Zuosha rt Printed by the Standards Bureau of the Ministry of Economic Affairs' ---- Cooperatives Printed Together 404951 A7 B7 V. Description of Invention (W)

CAC CAG CAG TGC TGT GAA AAG TGG ACC CTG GTT AAT GAT GAG ACC CAA GCC AAG 4509 4536

His Gin Gin Cys Cys Glu hys Trp Thr Leu Val Ann Asp Π3υ Thr Gin Λίη? .Yn 1495 1500 1505 .1510

ATG GCC CGG ATG GCT GCT GCA GCT GCA TGG GGT TTA GGT CAG TGG GAC AGC ATG 4563 4590

MET Ala Arg MET Ala Ala Ala Ala Ala Trp Gly Leu Gly Gin Trp Asp Ser MET 1515 1520 1525

GAA GAA TAC ACC TGT ATG ATC CCT CGG GAC ACC CAT GAT GGG GCA TTT TAT AGA 4617 4644

Glu Glu Tyr Thr Cys MET lie Pro Arg Asp Thr His Asp Gly Ala Phe Tyr Arq 1530 1535 1540 1545

GCT GTG CTG GCA CTG CAT CAG GAC CTC TTC TCC TTG GCA CAA CAG TGC ATT GAC 4671 4698

Ala Val Leu Ala Leu Hie Gin Asp Leu Phe Ser. Leu Ala Gin Gin Cys lie Asp 1550 1555 1560 1565

AAG GCC AGG GAC CTG CTG GAT GCT GAA TTA ACT GCA ATG GCA GGA GAG TAC 4725 4752

Lys Ala Arg Asp Leu Leu Asp Ala Glu Leu Thr Ala MET Ala Gly Glu Ser Tyr 1570 1575 1580

AGT CGG GCA TAT GGG GCC ATG GTT TCT TGC CAC ATG CTG TCC GAG CTG GAG GAG 4779 4806

Ser Arg Ala Tyr Gly Ala MET Val Ser Cys His MET Leu Ser Glu I # eu Glu Glu 1585 1590 1595 1600

GTT ATC CAG TAC AAA CTT GTC CCC GAG CGA CGA GAG ATC ATC CGC CAG ATC TGG 4833 4860

Val lie Gin Tyr Lys Leu Val Pro Glu Arg Arg Glu lie lie Arg Gin lie Trp 1605 1610 1615

TGG GAG AGA CTG CAG GGC TGC CAG CGT ATC GTA GAG GAC TGG CAG AAA ATC CTT 4887 4914

Trp Glu Arg Leu Gin Gly Cys Gin Arg lie Val Glu Asp Trp Gin Lys He Leu 1620 1625 1630 1635

ATG GTG CGG TCC CTT GTG GTC AGC CCT CAT GAA GAC ATG AGA ACC TGG CTC AAG 4941 4968 MET Val Arg Ser Leu Val Val Ser Pro His Glu Asp MET Arg Thr Trp Leu Lys 1640 1645 1650 1655

TAT GCA AGC CTG TGC GGC AAG AGT GGC AGG CTG GCT CTT GCT CAT AAA ACT TTA 4995 5022

Tyr Ala Ser Leu Cys Gly Lys Ser Gly Arg Leu Ala Leu Ala His Lys Thr Leu 1660 1670

GTG TTG CTC CTG GGA GTT GAT CCG TCT CGG CAA CTT GAC CAT CCT CTG CCA ACA * 5049 5076

Val Leu Leu Leu Gly Val Asp Pro Ser Arg Gin Leu Asp His Pro Leu Pro Thr 1675 1680 1685 1§90 -45- This paper size is applicable to Chinese families (CNS > M size (210X297)) I- -: ---- 1--installation-* ·. (Please read the “Precautions on the back page” first) Member of the Central Standards Bureau of the Ministry of Economic Affairs r: Cooperative Du printed installation 4〇495l λτ B7 V. Description of the invention ( Frame) GGA GAC GGT TTG GTG AAA CCA GAG GCC CTA AAT AAG AAA GCT ATC CAG ATT ΑΤΪ 7479 750 £

Gly Asp Gly Leu Val Lys Pro Glu Ala Leu Asn hys Lys Ala lie Gin lie Il € 2485 2490 2495 2500

AAC AGG GTT CGA GAT AAG CTC ACT GGT CGG GAC TTC TCT CAT GAT GAC ACT TTG 7533 7560

Asn Arg Val Arg Asp Lys Leu Thr Gly Arg Asp Phe Ser His Asp Asp Thr Leu 2505 2510 2515

GAT GTT CCA ACG CAA GTT GAG CTG CTC ATC AAA CAA GCG ACA TCC CAT GAA AAC 7587 7614

Asp Val Pro Thr Gin Val Glu Leu Leu lie Lys Gin Ala Thr Ser His Glu Asn 2520, 2525 2530 2535

CTC TGC CAG TGC TAT ATT GGC TGG TAC CCT TTC TGG TAA 7641

Leu Cys Gin Cys Tyr lie Gly Trp Tyr Pro Phe Trp 2540 2545 ⑶ (3 > Information of Sequence ID No. 2: (U sequence characteristics: (A) Spread: Amino acid (B) Type ·· C) Amineness: Meaning direction of double-stranded DNA strands (D) Topology: Straight line (Π) Molecular type: Meaning of double-stranded cDNA to · ΝΝΑ (A) Sequence number 2 illustrates some of the GST-SEP fusion proteins. Starts with a linker sequence between the GST and SEP45 portions of the protein (iiU hypothesis: None (iv) Antisense: None (vi) Source: (A) Organ: Molt 4 Human T-cell leukemia cell (B) Strain: ATCC Strain CRL 1582 Ui) Sequence description: SEQ.IDN0: 2 ATG TCC CCT ΑΤΑ CTA GGT TAT TGG AAA ATT AAG GGC CTT GTG CAA CCC ACT CGi 2Ί 54 MET Ser Pro He Leu Gly Tyr Trp Lys He Lys Gly Leu Val Gin Pro Thr Arc 15 10 15

CTT CTT TTG GAA TAT CTT GAA GAA AAA TAT GAA GAG CAT TTG TAT GAG CGC GAT 81 10 (

Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asj 20 25 30 35 Clothes and paper are standard for the Chinese Communist Party (CNS > A4 size (210X297))

4ϋ ^ Ιθ5χ Α7 Β7 V. Description of the invention (still) Printed by the Central Bureau of Standards of the Ministry of Economy R '·' Fee Cooperative printed by TGG CGA AAC AAA AAG TXT GAA TTG GGT TTG GAG TTT CCC AAT 135 162 Trp Arg Asn Lys Lys Phe Glu Leu Gly Leu Glu Phe Pro Asn 45 50 CTT CCT TAT TAT ATT GAT GGT GAT GTT AAA TTA ACA CAG TCT ATG GCC ATC ΛΤΛ 189 216 Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys Leu Thr Gin Ser MET Ala lie lie 55 60 65 70 CGT TAT ATA GCT GAC AAG CAC AAC ATG TTG GGT GGT TGT CCA AAA GAG CGT GCA 243 270 Arg Tyr lie Ala Asp Lys His Asn MET Leu Gly Gly Gly Cys Pro Lys Glu Arg Arg Ala 75 80 85 90 GAG ATT TCA ATG CTT GAA GGA GCG GTT TTG GAT ATT AGA TAC GGT GTT TCG AGA 297 3 * 24 Glu lie Ser MET Leu Glu Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg 95 100 105 ATT GCA TAT AGT AAA GAC TTT GAA ACT CTC AAA GTT GAT TTT CTT AGC AAG CTA 351 378 lie Ala Tyr Ser Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser 4 * ys Leu 110 115 120 125 CCT GAA ATG CTG AAA ATG TTC GAA GAT CGT TTA TGT CAT AAA ACA TAT TTA AAT 405 432 Pro Glu MET Leu Lys MET Phe Glu As p Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 GGT GAT CAT GTA ACC CAT CCT GAC TTC ATG TTG TAT GAC GCT CTT GAT GTT GTT 459 486 Gly Asp His Val Thr His Pro Asp Phe MET Leu Tyr Asp Ala Leu Asp Val Val 145 150 155 160 TTA TAC ATG GAC CCA ATG TGC CTG GAT GCG TTC CCA AAA TTA GTT TGT TTT AAA 513 540 Leu Tyr MET Asp Pro MET Cys Leu Asp Ala Phe Pro Lys Leu Val Cys Phe Lys 165 170 175 180 AAA CGT ATT GAA GCT ATC CCA CAA ATT GAT AAG TAC TTG AAA TCC AGC AAG TAT 567 594 Lys Arg He Glu Ala lie Pro Gin lie Asp Lys Tyr Leu Lys Ser Ser Lys Tyr 185 190 195 ATA GCA TGG CCT TTG CAG GGC TGG CAA GCC ACG TTT GGT GGT GGC GAC CAT CCT 621 648 lie Ala Trp Pro Leu Gin Gly Trp Gin Ala Thr Phe Gly Gly Gly Gly Asp His Pro 200 205 210 215 CCA AAA TCG GAT CTG GTT CCG CGT GGT GGA TCC CCG GGA ATT TCC GGT GGT GGT 675 702 Pro Lys Ser Asp Leu Val Pro Arg Gly Gly Ser Pro Pro Gly lie Ser Gly Gly Gly 220 225 230 GAA GGT GAT AAA Glu Gly Asp Lys 40 (please read it first and pay attention to it on this page) Deaf ·

• T-51- 404951 V. Description of the invention (卬) A7 B7

GGT GGT

ATT CTA GAC

Gly Gly Gly lie Leu Asp 235 240 GAC TCC ATG AGC 729 Asp Ser MET Ser AAG AGT GGC AGG CTG GCT CCT GCT Lys Ser Gly Arg Leu Ala Leu Ala 255 260 GAT CCG TCT CGG CAA CTT GAC CAT Asp Pro Ser Arg Gin Leu Asp His 275 CAT AAA 783 His Lys TAT GCC TAC ATG Tyr Ala Tyr MET 290 AAA AAC ATG TGG Lys Asn MET Trp 295 CCT CTG 837 Pro Leu 280 AAG AGT 891 Lys Ser TTC AAG TAT GCA AGC Phe Lys Tyr Ala Ser 245 ACT TTA GTG TTG CTC Thr Leu Val Leu Leu 265 CCA ACA GTT CAC CCT Pro Thr Val His Pro 285 GCC CGC AAG ATC GAT Ala Arg Lys lie Asp 300 CTG TGC GGC 756 Leu Cys Gly 250 CTG GGA GTT 810 Leu Gly Val 270 CAG GTG ACC 864 Gin Val Thr CAC ATG CAG CAT TTT GTC CAG ACC His MET Gin His Phe Val Gin Thr 310 ACT GAG GAC CAG CAG CAT Asp Gin

Thr Glu 325

Gin His 330 AAG CAG Lys Gin ATG CAG 945 MET Gin 315 GAA CTG 999 Glu Leu CAA CAG GCC CAG CAT Gin Gin Ala Gin His 320 CAC AAG CTC ATG GCC His Lys Leu MET Ala 335 GCC TTC CAG 918 Ala Phe Gin 305 GCC ATC GCT 972 Ala He Ala CTG AAA CTT GGA GAG TGG Leu Lys Leu Gly Glu Trp 345 CCC AAA Pro Lys GTG CTG CAG TAC Val Leu Gin Tyr 365 Printed by AAG GCC TGG CAT GCG TGG Lys Ala Trp His Ala Trp 380, CAT CAG AAC CAA GCC CGC His Gin Asn Gin Ala Arg 400 AAC ATC ACC AAC GCC ACC Asn lie Thr Asn Ala Thr 415 420 CAG CTG AAT CTA 1053 Gin Leu Asn Leu 350 TAC AGC GCC GCC 1107 Tyr Ser Ala Ala 370 GCA GTG ATG AAC 1161 Ala Val MET Asn 385 GAT GAG AAG AAG 1215 Asp Glu Lys Lys 405 ACT GCC GCC ACC 1269 Thr Ala Ala Thr CAG GGC ATC AAT GAG Gin Gly lie Asn Glu 355 ACA GAG CAC GAC CGC TGC Glu His Asp Arg 375 TTC GAA GCT GTG CTA Phe Glu Ala Val Leu 390 AAA CTG CGT CAT GCC Lys Leu Arg His Ala 410 ACX3 GCC GCC ACT GCC Thr Ala Ala Thr Ala 425 CGA TGC TTC 1026 Arg Cys Phe 340 AGC ACA ATC 1080 Ser Thr lie 360 AGC TGG TAC 1134 Ser Trp Tyr CAC TAC AAA 1188 His Tyr Lys 395 AGC GGG GCC 1242 Ser Gly Ala ACC ACC ACT 1296 Thr Thr Thr 430 --------- i ----- --- IT ------ m (please read the note ^ on the back before filling in this page) The paper size is applicable ** furniture ratio (CNS) A4 specification (210X297 mm) 404951 A7 B7 5 , Description of invention (fantasy)

GCC AGC ACC GAG GGC AGC AAC AGT GAG AGC GAG GCC GAG AGC ACC GAG AAC AGC 1323 1350

Ala Ser Thr Glu Gly Ser Asn Ser Glu Ser Glu Ala Glu Ser ihr Γϊΐη Ληη Γ.γ »γ 435 440 .445 Λ50

CCC ACC CCA TCG CCG CTG CAG AAG AAG GTC ACT GAG GAT CTG TCC AAA ACC CTC 1377 1404

Pro Thr Pro Ser Pro Leu Gin Lys Lys Val Thr Glu Asp Leu Ser hys Thr Leu 455 460 465

CTG ATG TAC ACG GTG CCT GCC GTC CAG GGC TTC TTC CGT TCC ATC TCC TTG TCA 1431 1458

Leu MET Tyr Thr Val Pro Ala Val Gin Gly Phe Phe Arg Ser lie Ser Leu Ser 470 475 480 485

CGA GGC AAC AAC CTC CAG GAT ACA CTC AGA GTT CTC ACC TTA TGG TTT GAT TAT 1485 1512

Arg Gly Asn Asn Leu Gin Asp Thr Leu Arg Val Leu Thr Leu Trp Phe Asp Tyr 490 495 500

GGT CAC TGG CCA GAT GTC AAT GAG GCC TTA GTG GAG GGG GTG AAA GCC ATC CAG 1539 " 1566

Gly His Trp Pro Asp Val Asn Glu Ala Leu Val Glu Gly Val Lys Ala lie Gin 505 510 515 520

ATT GAT ACC TGG CTA CAG GTT ATA CCT CAG CTC ATT GCA AGA ATT GAT ACG CCC 1593 1620 lie Asp Thr Trp Leu Gin Val lie Pro Gin Leu He Ala Arg lie Asp Thr Pro 525 530 535 540

AGA CCC TTG GTG GGA CGT CTC ATT CAC CAG CTT CTC ACA GAC ATT GGT CGG TAC 1647 1674

Arg Pro Leu Val Gly Arg Leu lie His Gin Leu Leu Thr Asp He Gly Arg Tyr 545 550 555

CAC CCC CAG GCC CTC ATC TAC CCA CTG ACA GTG GCT TCT AAG TCT ACC ACG ACA 1701 1728

His Pro Gin Ala Leu lie Tyr Pro Leu Thr Val Ala Ser Lys Ser Thr Thr Thr 560 565 570 575

GCC CGG CAC AAT GCA GCC AAC AAG ATT CTG AAG AAC ATG TGT GAG CAC AGC AAC Member of the Central Standards Bureau of the Ministry of Economic Affairs Printed by the Cargo Cooperative (谙 Read first. ** Footnote '* 洚 芩 #Write this page) 1755 1782

Ala Arg His Asn Ala Ala Asn Lys He Leu Lys Asn MET Cys Glu His Ser Asn 580 585 590

ACC CTG GTC CAG CAG GCC ATG ATG GTG AGC GAG GAG CTG ATC CGA GTG GCC ATC 1809 1836

Thr Leu Val Gin Gin Ala MET MET Val Ser Glu Glu Leu lie Arg Val Ala lie 595 600 605 610

CTC TGG CAT GAG ATG TGG CAT GAA GGC CTG GAA GAG GCA TCT CGT TTG TAC TTT 1863 1890

Leu Trp His Glu MET Trp His Glu Gly Leu Glu Glu Ala Ser Arg Leu Tyr Phe 615 620 625, 630 -53- Too much paper ruler for use · Zhongye storehouse friends choose who to come together, 0/7 grid, 404951 A7 B7 V. Description of the invention (^ GGG GAA Gly Glu AGG AAC Arg Asn ATG GAA CGG GGC MET Glu Arg Gly 650 CGA GAT TTA ATG Arg Asp I * eu MET 670 GTC AAG GAC CTC Val Lys Asp Leu 685 TCA AAG CAG CTG Ser Lys Gin Leu 705 CTT CTG Leu Leu ATG TGC MET Cys CAG CCA ATC ATT Gin Pro lie lie 740 GTG AAA GGC ATG TTT 1917 Val Lys Gly MET Phe 635 CCC CAG ACT CTG AAG 1971 Pro Gin Thr Leu Lys 655 GAG GCC CAA GAG TGG 2025 Glu Ala Gin Glu Trp 675 ACC CAA GCC TGG GAC 2079 Thr Gin Ala Trp Asp 690 CCT CAG CTC ACA TCC 2133 Pro Gin Leu Thr Ser 710 CGG GAC CTT GAA TTG 2187 Arg Asp Leu Glu Leu 725 CGC ATT CAG TCC ATA 2241 lie Gin Ser He 745 GAG GTG CTG GAG CCC TTG CAT GCT ATG 1944 Glu Val Leu Glu Pro Leii His Ala MET 640 645 GAA ACA TCC TTT AAT CAG GCC TAT GGT 1998 Glu Thr Ser Phe Asn Gin Ala Tyr Gly 660 665

TGC AGG AAG TAC ATG

TCA AAG CAG AGG CCC CGG Lys Gin

Arg Pro 760 h TTG ACA CTT 2295 Arg hys Leu Thr Leu T65 Member of the sample bureau in the Ministry of Economic Affairs u 'Μ cooperation Du Yin «. TTC CTT CTA AAA Phe Leu Leu Lys 775 GGC CAT GAA GAT CTG 2349 Gly His Glu Asp Leu 780 TTC GGC CTG GTT AAC ACC CTT CTG GCC 2403 Phe Gly Leu Val Asn Thr Leu Leu Ala 795 800CTC AGC ATC CAG AGA TAC GCT GTC ATC 2457Leu Ser lie Gin Arg Tyr Ala Val lie 815; AAT 2052 Cys Arg Lys Tly MET Lys Ser Asn 680 CTC TAT TAT CAT GTG TTC CGA CGA ATC 2106 Leu Tyr Tyr His Val Phe Arg Arg lie 695. 700 TTA GAG CTG CAA TAT GTT TCC CCA AAA 2160 Leu Glu Leu Gin Tyr Val Ser Pro Lys 715 720 GCT GTG CCA GGA ACA TAT GAC CCC AAC 2214 Ala Val Pro Gly Thr Tyr Asp Pro Asn 730 735 GCA CCG TCT TTG CAA GTC ATC ACA TCC 2268 Ala Pro Ser Leu Gin Val lie Thr Ser 750 755 ATG GGC AGC AAC GGA CAT GAG TTT GTT 2322 MET Gly Ser Asn Gly His Glu Phe Val 770 CGC CAG GAT GAG CGT GTG ATG CAG CTC 2376 Arg Gin Asp Glu Arg Val MET Gin Leu 785 790 AAT GAC CCA ACA TCT CTT CGG AAA AAC 2430 Asn Asp Pro Thr Ser Leu Arg Lys Asn 805 810 CCT TTA TCG ACC AAC TCG GGC CTC ATT 2484 Pro Leu Ser Thr Asn Ser Gly Leu lie 820 825 items-• 54 · This paper size uses two samples from China · (CNS > Λ4 size (210X297 mm) 404951 A7 B7 5. Description of the invention

GGC TGG GTT CCC CAC TGT GAC ACA CTG CAC GCC CTC ATC CGG GAC TAC AGG GAG 2511 2538

Gly Trp Val Pro His Cys Asp Thr Leu His Ala Leu lie Arg Asp Tyr Arg Glu 830 835 840 845

AAG AAG AAG ATC CTT CTC AAC ATC GAG CAT CGC ATC ATG TTG CGG ATG GOT CCG 2565 2592

Lys Lys Lys lie Leu Leu Asn lie Glu His Arg lie MET Leu Arg MET Ala Pro 850 855 860

GAC TAT GAC CAC TTG ACT CTG ATG CAG AAG GTG GAG GTG TTT GAG CAT GCC GTC 2619 2646

Asp Tyr Asp His Leu Thr Leu MET Gin Lys Val Glu Val Phe Glu His Ala Val 865 870, 875 880

AAT AAT ACA GCT GGG GAC GAC CTG GCC AAG CTG CTG TGG CTG AAA AGC CCC AGC 2673 2700

Asn Asn Thr Ala Gly Asp Asp Leu Ala Lys Leu Leu Trp Leu Lys Ser Pro Ser 885 890 895 900

TCC GAG GTG TGG TTT GAC CGA AGA ACC AAT TAT ACC CGT TCT TTA GCG GTC ATG 2727 — 2754

Ser Glu Val Trp Phe Asp Arg Arg Thr Asn Tyr Thr Arg Ser Leu Ala Val MET 905 910 915

TCA ATG GTT GGG TAT ATT TTA GGC CTG GGA QAT AGA CAC CCA TCC AAC CTG ATG 2781 2808

Ser MET Val Gly Tyr lie Leu Gly Leu Gly Asp Arg His Pro Ser Asn Leu MET 920 925 930 935

CTG GAC CGT CTG AGT GGG AAG ATC CTG CAC ATT GAC TTT GGG GAC TGC TTT GAG 2835 2862

Leu Asp Arg Leu Ser Gly Lys lie Leu His lie Asp Phe Gly Asp Cys Phe Glu 940 945 950

GTT GCT ATG ACC CGA GAG AAG TTT CCA GAG AAG ATT CCA TTT AGA CTA ACA AGA 2889 2916

Val Ala MET Thr Arg Glu Lys Phe Pro Glu Lys lie Pro Phe Arg Leu Thr Arg 955 960 965 970

ATG TTG ACC AAT GCT ATG GAG GTT ACA GGC CTG GAT GGC AAC 'TAC AGA ATC ACA 2943 2970 MET Leu Thr Asn Ala MET Glu Val Thr Gly Leu Asp Gly Asn Tyr Arg lie Thr 975 980 985 990

TGC CAC ACA GTG ATG GAG GTG CTG CGA GAG CAC AAG GAC AGT GTC ATG GCC GTG 2997 3024

Cys His Thr Val MET Glu Val Leu Arg Glu His Lys Asp Ser Val MET Ala Val 995 1000 1005

CTG GAA GCC TTT GTC TAT GAC CCC TTG CTG AAC TGG AGG CTG ATG GAC ACA AAT 3051 3078

Leu Glu Ala Phe Val Tyr Asp Pro Leu Leu Asn Trp Arg Leu MET Asp Thr Asn 1010 1015 1020 1025 This paper size applies to the China National Standard (CNS > A4 specification (210X297 mm) —.1 I — I — II- , I — order — —— I I- ^ (Please read the ^^^ service on the back before filling this page) Printed by the Male Workers Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economics --- B-7404951 V. Description of Invention (^ ) ACC AAA GGC AAC AAG CGA TCC CGA ACG AGG ACG GAT TCC TAC TCT GCT GGC CAG 3105 3132 Thr Lys Gly Asn Lys Arg Ser Arg Thr Arg Thr Asp Ser Tyr Ser Ala Gly Gin 1030 1035 1040 TCA GTC GAA ATT TTG GAC GGT GTG GAA CTT GGA GAG CCA GCC CAT AAG AAA ACG 3159 3186 Ser Val Glu lie Leu Asp Gly Val Glu Leu Gly Glu Pro Ala His Lys Lys Thr 1045 1050 1055 1060 GGG ACC ACA GTG CCA GAA TCT ATT CAT TCT TTC ATT GGA GAC GGT TTG GTG AAA 3213 3240 Gly Thr Thr Val Pro Glu Ser lie His Ser Phe lie Gly Asp Gly Leu Val Lys 1065 1070 1075 1080 CCA GAG GCC CTA AAT AAG AAA GCT ATC CAG ATT ATT AAC AGG GTT CGA GAT AAG 3267 3294 Pro Glu Ala Leu Asn Lys Lys Ala He Gin lie lie Asn Arg Val Arg Asp Lys 1085 1090 '1095 CTC ACT GGT CGG GAC TTC TCT CAT GAT GAC ACT TTG GAT GTT CCA ACG CAA GTT 3321 3348 Leu Thr Gly Arg Asp Phe Ser His Asp Asp Thr Leu Asp Val Pro Thr Gin Val 1100 1105 1110 1115 GAG CTG CTC ATC AAA CAA GCG ACA TCC CAT GAA AAC CTC TGC CAG TGC TAT ATT 3375 3402 Glu Leu Leu lie Lys Gin Ala Thr Ser His Glu Asn Leu Cys Gin Cys Tyr lie 1120 1125 1130 GGC TGG TAC CCT TTC TAA Gly Trp Tyr Pro Phe Trp 1135 1140 --------- d -------- ΐτ ------ ^ (Please read the back ^ aE-v before filling this page) Economy The paper size of the printed paper of the Central Bureau of Standards and Commerce of the People's Republic of China is applicable to China® Furniture Standard (CNS) A4 (210X297 mm)

Claims (1)

Central Standards Bureau of the Ministry of Economy It encodes a part of human-derived peptone that can be bound to the GST-FKBP-rapamycin complex, and the sequence is as follows: AAG ATG CTT GGA ACC GGA CCT GCC GCC GCC ACC ACC GCT GCC ACC ACA TOT AGC 27 54 Met Leu Gly Thr Gly Pro Ala Ala Ala Thr Thr Ala Ala Thr Thr Ser Ser 15 10 15 AAT GTG AGC GTC CTG CAG CAG TTT GCC AGT GGC CTA AAG AGC CGG AAT GAG GAA 81 108 Asn Val Ser Val Leu Gin Gin Phe Ala Ser Gly Leu Lys Ser Arg Asn Glu Glu »20 25 30 35 ACC AGG GCC AAA GCC GCC AAG GAG CTC CAG CAC TAT GTC ACC ATG GAA CTC CGA 135 162 Thr Arg Ala Lys Ala Ala Lys Glu Leu Gin His Tyr Val Thr MET Glu Leu Arg 40 45 50 GAG ATG AGT CAA GAG GAG TCT ACT CGC TTC TAT GAC CAA CTG AAC CAT CAC ATT 189 216 Glu MET Ser Gin Glu Glu Ser Thr Arg Phe Tyr Asp Gin Leu Asn His His lie 55 60 65 70 TTT GAA TTG GTT TCC AGC TCA GAT GCC AA T GAG AGG AAA GGT GGC ATC TTG GCC 243 270 Phe Glu Leu Val Ser Ser Ser Asp Ala Asn Glu Arg Lys Gly Gly lie Leu Ala 75 80 85 ATA GCT AGC CTC ATA GGA GTG GAA GGT GGG AAT GCC ACC CGA ATTjGGC AGA. TTT 297 324 He Ala Ser Leulie Gly Val Glu Gly Gly Asn Ala Thr Arg He Gly Arg Phe 90 95 100 105 GCC AAC TAT CTT CGG AAC CTC CTC CCC TCC AAT GAC CCA GTT GTC ATG GAA ATG 351 378 Ala Asn Tyr Leu Arg Asn Leu Leu Pro Ser Asn Asp Pro Val Val MET Glu MET 110 115 120 125 GCA TCC AAG GCC ATT GGC CGT CTT GCC ATG GCA GGG GAC ACT TTT ACC GCT GAG 405 432 Ala Ser Lys Ala lie Gly Arg Leu Ala MET Ala Gly Asp Thr Phe Thr Ala Glu 130 135 140 TAC GTG GAA TTT GAG GTG AAG CGA GCC CTG GAA TGG CTG GGT GCT GAC CGC AAT 459 486 Tyr Val Glu Phe Glu Val Lys Arg Ala Leu Glu Trp Leu Gly Ala Asp Arg Asn 145 150 155 160 Sheet The scale is applicable to the rate of the middle and poor countries (CNS) A4 (210X297 mm) in ^ — n ^ in ^ i 1 ^ 1 im III— —I— —f ^ in ^ —-(Please read the note on the back first ^ • Fill in this page again) It QC 4 A, B1C, D Application for patents Fanyuan GAG CGG AGA CAT Glu Gly Arg Arg His 165 GCA GCT Ala Ala CCT ACC TTC TTC TTC CAG CAA Pro Thr Phe Phe Phe Gin Gin Work 185 GTG TGG GAC CCC AAA CAG GCC Val Trp Asp Pro Lys Gin Ala 200 TGT CTG ATT CTC ACA ACC CAG Cys Leu He Leu Thr Thr Gin 220 TAC AGG CAC ACA TTT GAA GAA Tyr Arg His Thr Phe Glu Glu 235 240 GAG AAG GGC ATG AAT CGG GAT Glu Lys Gly MET Asn Arg Asp 255 GAG CTG GTC CGA ATC AGC AGC Glu Leu Val Arg lie Ser Ser 270 275GAA ATC ACA CAG CAG CAG CTG Giu lie Thr Gin Gin Gin Leu 290 GTC CTG 513 Val Leu 170 GTG CAA 567 Val Gin GTT CTC CGT GAG Val lieu Arg Glu CCC TTC TTT GAC Pro Phe Phe Asp 190 ATC CGT 621 lie Arg 205 CGT GAG 675 Arg Glu GAG GGA Glu Gly GCT GTA Ala Val 210 GCA GAG 729 Ala Glu CCG AAG GAG ATG Pro Lys Glu MET 225 AAG GGA TTT GAT Lys Gly Phe Asp 245 CTG GCC ATC AGC GTC 540 Leu Ala lie Ser Val 175 AAC ATT TTT GTG GCC 594 Asn lie Phe Val Ala 195 GCC GCC CTT CGT GCC 648 Ala Ala lieu Arg Ala 215 CAG AAG CCT CAG TGG 702 Gin Lys Pro Gin Trp 230 GAG ACC TTG GCC AAA 756 Glu Thr Leu Ala Lys 250 GAT CGG ATC CAT GGA GCC TTG TTG ATC'CTT AAC 783 810 Asp Arg lie His Gly Ala Leu Leu lie Leu Asn 260 265 ATG GAG 837 MET Glu GTA CAC 891 Va丄 His 295 GGA GAG CGT CTG Gly Glu Arg Leu 280GAC AAG TAC TGC Asp Lys Tyr Cys 300 AGA GAA GAA ATG GAA 864 Arg Glu Glu MET Glu -2-65 / AAA GAT CTC ATG GGC 918 Lys Asp Leu McT Gly 305 HI 1 ^ 1 11 —a— in ^^^ 1 I (Please read the precautions on the back before filling in this page) Order printed by the Central Bureau of the Ministry of Economic Affairs, Shellfish Consumer Cooperatives TTC GGA ACA AAA CCT CGT CAC ATT ACC CCC TTC ACC AGT TTC CAG GCT GTA CAG 945 972 Phe Gly Thr Lys Pro Arg His lie Thr Pro Phe Thr Ser Phe Gin Ala Val Gin 310 315 320 CCC CAG CAG TCA AAT GCC TTG GTG GGG CTG CTG GGG TAC AGC TCT CAC CAA GGC 999 1026 Pro Gin Gin Ser Asn Ala Leu Val Gly Leu Leu Gly Tyr Ser Ser His Gin Gly 325 330 335 340 CTC ATG GGA TGA GTT GG ACC TCC CCC AGT CCA GCT AAG TCC ACC CTG GTG GAG AGC 1053 1080 Leu MET Gly Phe Gly Thr Ser Pro Ser Pro Ala Lys Ser Thr Leu Val G lu Ser 345 350 355 This paper uses China National Standards (CNS > A4 Washing (210X297mm)) 404951 Application for patent scope A8 B8 C8 D8 CGG TGT TGC Arg Cys Cys 360 AGA GAC TTG ATG Arg Asp CTG TGC Leu Lys Cys 380 AGG AAT Arg Asn TTG CCC CGC TTG GCT Leu Pro Arg Leu Ala 400 Leu MET 365 AGC AAG Ser Lys GCA TTC Ala Phe GAG GAG 1107 Glu Glu AAC TCG 1161 Asn Ser 385 CGA CCT 1215 Arg Pro Lys Phe 370 CTG ATC Leu lie TTT GAT CAG Asp Gin CAA ATG Gin MET 390 GTG TGC Val Cys ACA ATC Thr lie TCT GCC TTC ACA GAT ACC Phe Thr Ser Ala 405 GAT ACC ATG AAC CAT GCC MET Asn Gin Asp Thr 415 His Ala 420 CTA AGC 1269 Leu Ser TGT GTC AAG AAG Cys Val Lys Lys 425 Asp Thr 410 GAG AAG Glu Lys CAG TGG GTG 1134 Gin Trp Val 375 CTT AAT TTG 1188 Leu Asn Leu 395 CAG TAT CTC 1242 Gin Tyr Leu GAA CGT ACA 1296 Glu Arg Thr 430 GCG GCC TTC Ala Ala Phe CAA GCC Gin Ala 435 CTG GGG Leu Gly GTC TAT TTG CCT CGC GTG CTG Pro Arg Val Tyr Leu 450 Val Leu 455 CTA CTT 1323 Leu Leu 440 GAC ATC 1377 Asp l ie TCT GTG GCT GTG AGG TCT Ser Val Ala Val ATC CGA GCG GCC lie Arg Ala hla 460 Arg Ser 445 CTG CCC Leu Pro GAG TTT AAG 1350 Glu Phe Lys CCA AAG GAC 1404 Pro Lys Asp 465 n In In ^^ 1 l ^ i ϋ— ϋ— · ϋ 4 In nn 1 I — (please note the first item on Min Nian before filling this page) TTC GCC CAT AAG AGG CAG AAG Phe Ala His Lys Arg Gin Lys 470 GCA ATG 1431 Ala MET 475 CAG GTG GAC GCC ACA GTC Gin Val Asp Ala Thr Val 480 TTC ACT TGC 1458 Phe Thr Cys 〆-485 ATC AGC ATG CTG GCT CGA GCA lie Ser MET Leu Ala Arg Ala 490 ATG GGG1485 MET Giy CCA GGC ATC CAG CAG GAT lie Gin Pro Gly 495 CTG CTG GAG CCC ATG CTG GCA Pro MET 5-member Leu lieu Glu 505 TAG GAC CTG Tyr Asp Leu Leu Ala 510 GTG GGA 1539 Val Gly CTA AGC CCT GCC Leu Ser Pro Ala 515 Gin Asp 500 CTC ACT Leu Thr ATC AAG GAG, 1512 lie Lys Giu AGC CGT CAG ATT Gin lie Ser Arg 525 CCA. CAG CTA AAG 1593 Pro Gin Leu Lys 530 AAG GAC ATT Lys Asp lie Gin S35 GCA GTG CTC 1566 Ala Val Leu 520 GAT GGG CTA 1620 A sp Gly Leu CTG AAA ATG CTG TCC CTG GTC Leu Ser Leu Lys MET 540 Leu Val 545 CCC AAG GGC CTG GCC CAT CAG CTT ATG CAC AAA 1647 Leu MET His Lys 550 CTG GCC TCT CCT 1701 CCC CTT Pro Leu CGC CAC CCA Arg His ^ ATG 1674 Pxo Gly MET 555 CTC ACG ACC CTC CCT GAG 1728 -v 'Paper ruler A is in use «National rate (CNS) A4 washing grid (210X297 mm) A8404951 c | Printed by the staff consumer cooperative of the Bureau Pro Lys Gly Leu Ala His Gin Leu Ala Ser Pro Gly Leu Thr Thr Leu Pro Glu 560 565 570 575 GCC AGC GAT GTG GGC AGC ATC ACT CTT GCC CTC CGA ACG CTT GGC AGC TTT GAA 1755 1782 Ala Ser Asp Val Gly Ser lie Thr Leu Ala Leu Arg Thr Leu Gly Ser Phe Glu 580 585 590 TTT GAA GGC CAC TCT CTG ACC CAA TTT GTT CGC CAC TGT GCG GAT CAT TTC CTG 1809 1836 Phe Glu Gly His Ser Leu Thr Gin Phe Val Arg His Cys Ala Asp His Phe hen 595 600 605 610 AAC AGT GAG CAC AAG GAG ATC CGC ATG GAG GCT GCC CGC ACC TGC TCC CGC CTG 1863 1890 Asn Ser Glu His Lys Glu lie Arg MET Glu Ala Ala Arg Thr Cys Ser Arg Leu 615 620 € 25 CTC ACA CCC TCC ATC CAC CTC ATC AGT GGC CAT GCT CAT GTG GTT AGC CAG ACC 1917 1944 Leu Thr Pro Ser He His Leu lie Ser Gly His Ala His Val Val Ser Gin Thr 630 635 640 645 645 GCA GTG CAA GTG GTG GCA GAT GTG CTT AGC AAA CTG CTC GTA GTT GGG ATA ACA 1971 1998 Ala Val Gin Val Val Ala Asp Val Leu Ser Lys Leu Leu Val Val Gly -lie Thr 650 655 660 665 GAT CCT GAC CCT GAC ATT CGC TAC TGT GTC TTG GCG TCC CTG GAC GAG CGC TTT 2025 2052 Asp Pro Asp Pro Asp lie Arg Tyr Cys Val Leu Ala Ser Leu Asp Glu Arg Phe 670 675 680 GAT GCA CAC CTG GCC CAG GCG GAG AAC TTG CAG GCC TTG TTT GTG GCT CTG AAJ, 2079 2106 Asp Ala His Leu Ala Gin Ala Giu Asn Leu Gin Ala Leu Phe Val Ala Leu Asn 685 690 695 700 GAC CAG GTG TTT GAG ATC CGG GAG CTG GCC ATC TGC ACT GTG GGC CGA CTC AGT 2133 2160 Asp Gin Phe Glu lie Arg Glu Leu Ala lie Cys Thr Val Gly Arg Leu Ser 705 710 715 AGC ATG AAC CCT GCC TTT GTC ATG CCT TTC CTG CGC AAG ATG CTC ATC CAG ATT 2187 2214 Ser MET Asn Pro Ala Phe Val MET Pro Phe Le u Arg Lys MET Leu lie Gin lie 720 725 730 735 TTG ACA GAG TTG GAG CAC AGT GGG ATT GGA AGA ATC AAA GAG CAG AGT GCC CGC 2241 2268 Leu Thr Glu Leu Glu His Ser Gly lie Gly Arg lie Lys Glu Gin Ser Ala Arg 740 745 750 755 ATG CTG GGG CAC CTG GTC TCC AAT GCC CCC CGA CTC ATC CGC CCC TAC ATG GAG 2295 2322 MET Leu Gly His Leu Val Ser Asn Ala Pro Arg Leu lie Arg Pro Tyr MET Glu 760 765 770! — — — — — Order — 11.— 11 4 (Please read the note above and then fill out this page) 彖 The paper size is applicable to China® Standard (CNS) A4 (2 丨 0X297mm) 404951 Patent Application A8 B8 C8 D8 CCT ATT CTG AAG GCA TTA ATT Lys Ala Pro lie Leu 775 CCA GGT GTG ATC AAT Pro Gly Val GGC CTG GAA Gly Leu Glu 810 lie Asn 795 ATG AGG MET Arg CTC CAG GAT TCC TCT Leu Gin Asp Ser Ser 830 CAG TTG GTG GCC AGC Gin Leu Val Ala Ser 850 Leu lie 780 AAT GTC Asn Val AAA TGG Lys Trp 815 TTG TTG Leu Leu ACT GGC Thr Gly TTG AAA 2349 Leu Lys CTG GCA 2403 Leu Ala 800 GTT GAT 2457 Val Asp CTG AAA Leu Lys ACA ATA Th r lie GAT CCA GAC CCT Asp Pro Asp Pro 785 GGA GAA Gly Glu GAA CTT TTT ATT Phe He GCC AAA 2511 Ala Lys 835 TAT GTA 2565 Tyr Val Glu Leu 820 AGG CAG Arg Gin TTG GCA Leu Ala 805 ATC ATC He lie GAT CCA AAC 2376 Asp Pro Asn 790 CAG GTT AGT 2430 Gin Val Ser GTG GCT CXG TGG Val Ala Leu Trp 840 TTG CTT GAG GTG CTA CTG AAT Val Leu Leu Leu Glu 865 AGA GAG GCC ATC Arg Glu Ala lie Arg 885 Leu Asn 870 GTG TTA Vai Leu TTT CTG 2619 Phe Leu GTA GAG CCC TAC AGG AAG Val Glu Pro Tyr Arg Lys 855 860 AAG ACT GAG CAG AAC CAG Lys Thr Glu Gin Asn Gin 875 ATG GAC ATG 2484 MET Asp MET 825 ACC CTG GGA 2538 Thr Leu Gly TAC CCT ACT 2592 Tyr Pro Thr GTG AAC ATT GGC ATG ATA He Gly Lys Val Asn 900 MET lie 905 GGG GTT 2673 Giy Leu 890 GAC CAG 2727 Asp Gin Printed by the Consumer Affairs Cooperative of the Central Government Bureau of the Ministry of Economic Affairs *. TCA GAA TCC AAG TCA Ser Glu Ser Lys Ser 920 GTC AAC ATG GGA AAC Val Asn MET Gly Asn 940 GCC CTG ATG CGG ATC Ala Leu MET Arg lie 955 GTC CAG GCC ATC ACC AGT CAG Ser Gin TTG CCT Leu Pro GAT TCC 2781 Asp Ser 925 CTG GAT 2835 Leu Asp TTA GGG GCT TTG Leu Gly Ala Leu TCC CGG GAT GCC Ser Arg Asp Ala 910 TCT GAC TAT AGC Ser Asp Tyr Ser 930 TTC CGA GAC CAG 2889 Phe Arg Asp Gin 960 TTC ATC TTC AAG 2943 GAT CCT Asp Pro 895 TCT GCT Ser Ala ACT AGT Thr Ser GGT ACA CGC 2646 Gly Thr Arg 880 ^ TAC AAG CAC 2700 Tyr Lys His nii I— 1 ^ 1--i- I--a I--.. .II i (please read 1 ^ side note $ Item before #write this page) GAG TTC TAC CCA GCT GTG Tyr Pro Glu Phe 945 Ala Val 950 GTC AGC CTG 2754 Val Ser Leu 915 GAA ATG CTG 2808 Glu MET Leu 935 TCC ATG GTG 2862 Ser MET Val TCA CTC TCT CAT Ser Leu Ser His 965 TCC CTG GGA CTC CAT CAC His His AAA TGT ACC ATG GTT 2916 Thr MET Val 970 GTG CAG TTC 2970 Sheet paper size (CNS M4) Regulations (Grid) (210X297 mm) 404951 A8 B8 C8 D8 Patent Application Scope of the Ministry of Economic Affairs, Central Government Bureau βζ 工 工 工 合作社 合作 印 *. Val Gin Ala lie Thr Phe lie Phe Lys Ser Leu Gly I ^ eu Lys Cys Val Gin Phe 975 980 985 CTG CCC CAG GTC ATG CCC ACG TTC CTT AAT GTC ATT CGA G TC TGT GAT GGG GCC 2997 3024 Leu Pro Gin Val MET Pro Thr Phe Leu Asn Val lie Arg Val Cys Asp Gly Ala 990 995 1000 1005 ATC CGG GAA TTT TTG TTC CAG CAG CTG GGA ATG TTG GTG TCC XTT GTG AAG AGC 3051 3078 lie Arg Glu Phe Leu Phe Gin Gin Leu Gly MET Leu Val Ser Phe Val Lys Ser 1010 1015 1020 1025 CAC ATC AGA CCT TAT ATG GAT GAA ATA GTC ACC CTC ATG AGA GAA TTC TGG GTC 3105 3132 His lie Arg Pro Tyr MET Asp Glu lie Val Thr Leu MET Arg Glu Phe Trp Val 1030 1035 1040 ATG AAC ACC TCA ATT CAG AGC ACG ATC ATT CTT CTC ATT GAG CAA ATT GTG GTA 3159 3186 MET Asn Thr Ser lie Gin Ser Thr lie He Leu Leu lie Glu Gin lie Val Val 1045 1050 1055 1060 GOT CTT GGG GGT GAA TTT AAG CTC TAC CTG CCC CAG CTG ATC CCA CAC ATG CTG 3213 3240 Ala Leu Gly Gly Glu Phe Lys Leu Tyr Leu Pro Gin Leu lie Pro Hi§, MET Leu 1065 1070 1075 " CGT GTC TTC ATG CAT GAC AAC AGC CCA GGC CGC ATT GTC TCT ATC AAG TTA CTG 3267 3294 Arg Vai Phe MET His Asp Asn Ser Fro Gly Arg lie Vai Ser iie i > ys Leu Leu 1080 1085 1090 1095 GCT GCA A TC CAG CTG TTT GGC GCC AAC CTG GAT GAC TAC CTG CAT TTA CTG CTG 3321 3348 Ala Ala lie Gin Leu Phe Gly Ala Asn Leu Asp Asp Tyr Leu His Leu Leu Leu 1100 1105 1110 ms CCT CCT ATT GTT AAG TTG TTT GCT CCC GAA GCT CCA CTG CCA TCT CGA AAG 3375 3402 Pro Pro lie Val Lys Leu Phe Asp Ala Pro Glu Ala Pro Leu Pro Ser Arg Lys 1120 1125 1130 GCA GCG CTA GAG ACT GTG GAC CGC CTG ACG GAG TCC CTG GAT TTC ACT GAC TAT 3429 3456 Ala Ala Leu Glu Thr Val Asp Arg Leu Thr Glu Ser Leu Asp Phe Thr Asp Tyr 1135 1140 1145 1150 GCC TCC CGG ATC ATT CAC CCT ATT GTT CGA ACA CTG GAC CAG AGC CCA GAA CTG 3483 3510 Ala Ser Arg lie lie His Pro lie Val Arg Thr Leu Asp Gin Ser Pro Glu Leu 1155 1160 1165 CGC TCC ACA GCC ATG GAC ACG CTG TCT TCA CTT GTT TTT CAG CTG GGG AAG AAG 3537 3564 Arg Ser Thr Ala MET Asp Thr Leu Ser Ser Leu Val Phe Gin I * eu Gly Lys Lys 1170 1175 1180 1185 m 1_1 til «an 1. ^ 1 m HI HI I in am HI--(please note the first $ item before filling in this page) This paper is scaled to those in poor countries (CNS) A4 Washer (210X297 male (%) 4〇4 A8 B8 C8 D8 VI. Application for Patent Scope Printed by the Ministry of Economic Affairs and the Central Bureau of Motor Vehicles, Shellfish Consumer Cooperatives *. TAC CAA ATT TTC ATT CCA ATG GTG AAT AAA GTT CTG GTG CGA CAC CGA ATC AAT 3591 3618 Tyr Gin lie Phe He Pro MET Val Asn hys Val Leu Val Arg His Arg lie Asn 1190 1195 1200 1205 CAT CAG CGC TAT GAT GTG CTC ATC TGC AGA ATT GTC AAG < 3GA TAC ACA CTT GOT 3645 3672 His Gin Arg Tyr Asp Val Leu lie Cys Arg lie Val Lys Gly Tyr Thr Leu Ala 1210 1215 1220 GAT GAA GAG GAG GAT CCT TTG ATT TAC CAG CAT CGG ATG CTT AGG AGT GGC CAA 3699 3726 Asp Glu Glu Glu Asp Pro Leu lie Tyr Gin His Arg MET Leu Arg Ser Gly Gin 1225 1230 1235 1240 GGG GAT GCA TTG GCT AGT GGA CCA GTG GAA ACA GGA CCC ATG AAG AAA CTG CAC 3753 3780 Gly Asp Ala Leu Ala Ser Gly Pro Val Glu Thr Gly Pro MET Lys Lyu Leu His 1245 1250 1255 GTC AGC ACC ATC AAC CTC CAA AAG GCC TGG GGC GCT GCC AGG AGG GTC TCC AAA 3807 3834 Val Ser Thr lie Asn Leu Gin Lys Ala Trp Gly Ala Ala Arg Arg Va ^ l Ser ^ Lys 1260 1265 1270 Γ275 GAT GAC TGG CTG GAA TGG CT G AGA CX3G CTG AGC CTG GAG CTG CTG AAG GAC TCA 3861 3888 Asp Asp Trp Leu Giu Trp Leu Arg Arg Leu Ser Leu Glu Leu Leu Lys ksp Ser 1280 1285 1290 1295 TCA TCG CCC.TCC CTG CGC TCC TGC TGG GCC CTG GCA GCC TAC AAC CCG ATG 3915 3942 Ser Ser Pro Ser Leu Arg Ser Cys Trp Ala Leu Ala Gin Ala Tyr Asn Pro MET 1300 1305 1310 GCC AGG GAT CTC TTC AAT GCT GCA TTT GTG TCC TGC TGG TCT GAA CTG AAT GAA 3969 3996 Ala Arg Asp Leu Phe Asn Ala Ala Phe Val Ser Cys Trp Ser Glu I # eu Asn Glu 1315 1320 1325 1330 GAT CAA CAG GAT GAG CTC ATC AGA AGC ATC GAG TTG GCC CTC ACC TCA CAA GAC 4023 4050 Asp Gin Gin Asp Glu Leu He Arg Ser lie Glu Leu Ala Leu Thr Ser Gin Asp 1335 1340 1345 ATC GCT GAA GTC ACA CAG ACC CTC TTA AAC TTG GCT GAA TTC ATG GAA CAC AGT 4077 4104 lie Ala Glu Val Thr Gin Thr Leu Leu Asn Leu Ala Glu Phe MET Glu His Ser 1350 1355 1360 1365 GAC AAG GGC CCC CTG CCA CTG AGA GAT GAC AAT GGC ATT GTT CTG CTG GGT GAG 4131 4158 Asp Lys Gly Pro Leu Pro Leu Arg Asp Asp Asn Gly lie Val Leu Leu Gly Glu 1370 1375 1375 380 1385 --------- ^ ------ tr ------ ^ (Please read the note on the f side before filling in this page) Standard (CNS) A4 now (210x297 mm) 404851 A8 B8 C8 DS 夂, patent application scope Central Standards Bureau of the Ministry of Economic Affairs β: printed by industrial and consumer cooperatives AGA GOT GCC AAG TGC CGA GCA TAT GCC AAA GCA CTA CAC TAC AAA GAA CTG GAG 4185 4212 Arg Ala Ala hys Cys Arg Ala Tyr Ala Lys Ala Leu His Tyr Lys Glu Leu Glu 1390 1395 1400 TTC CAG AAA GGC CCC ACC CCT GCC ATT CTA GAA TCT CTC ATC AGC ATT AAT AAX 4239 4266 Phe Gin Gly Pro Thr Pro Ala lie Leu Glu Ser Leu lie Ser lie A3n Asn 1405 1410 1415 1420 AAG CTA CAG CAG CAG CCG GAG GCA GCG GCC GGA GXG TTA GAA TAT GCC ATG AAA CAC 4293 4320 Lys Leu Gin Gin Pro Glu Ala Ala Ala Gly Val Leu Glu Tyr Ala MET Lys His 1425 1430 1435 TTT GGA GAG CTG GAG ATC CAG GCT ACC TGG TAT GAG AAA CTG CAC GAG TGG GAG 4347 4374 Phe Gly Glu Leu Glu lie Gin Ala Thr Trp Tyr Glu Lys Leu His Glu Trp Glu 1440 1440 1450 1455 GAT GCC CTT GTG GCC TAT GAC AAG AAA ATG GAC ACC AAC AAG GA C GAC CCA GAG 4401 4428 Asp Ala Leu Val Ala Tyr Asp Lys Lys MET Asp Thr Asn Lys Asp Asp Pro Glu 1460 1465 1470 〆〆 1475 CTG ATG CTG GGC CGC ATG CGC TGC CTC GAG GCC TTG GGG GAA TGG GGT GAA CTC * 4455 4482 Leu MET Leu Gly Arg MET Arg Cys Leu Glu Ala Leu Gly Glu Trp Gly Gin Leu '' 1480 1485 '1490 CAC CAG CAG TGC TGT GAA AAG TGG ACC CTG GTT AAT GAT GAG ACC CAA GCC AAG 4509 4536 His Gin Gin Cys Cys Glu Lys Trp Thr Leu Val Asn Asp Glu Thr Gin Ala Lys 1495 1500 1505 1510 ATG GCC CGG ATG GCT GCT GCT GCA GCT GCA TGG GGT TTA GGT CAG TGG GAC AGC ATG 4563 4590 MET Ala Arg MET Ala Ala Ala Ala Ala Trp Gly Leu Gin Trp Asp Ser MET 1515 1520 1S25 GAA GAA TAC ACC TGT ATG ATC CCT CGG GAC ACC CAT GAT GGG GCA TTT TAT AGA 4617 4644 Glu Glu Tyr Thr Cys MET lie Pro Arg Asp Thr His Asp Gly Ala Phe Tyr Arg 1530 1535 1540 GCT GTG CTG GCA CTG CAT CAG GAC CTC TTC TCC TTG GCA CAA CAG TGC ATT GAC 4671 4698 Ala Val Leu Ala Leu His Gin Asp Leu Phe Ser Leu Ala Gin Gin Cys lie Asp 1550 1555 1560 1565 AAG GCC AGG GAC CTG CTG GAT GCT GAA TTA ACT GCA ATG GCA GGA GAG AGT TAC 4725 4752 Lys Ala Arg Asp Leu Leu Asp Ala Glu Leu Thr Ala MET Ala Gly Glu Ser Tyr 1570 1575 1580 --------- ^ — (Please note the f item on the noodles page before filling in this page) This paper is used in China «Home Knitting Standard (CNS) Α4 Specification U〗 0X297 mm) 404951 a | C8 D8 VI. Patent Application Range AGT CGG GCA TAT GGG GCC ATG GTT TCT TGC CAC ATG CTG TCC GAG CTG GAG GAG 4779 4806 Ser Arg Ala Tyr Gly Ala MET Val Ser Cys His MET Leu Ser Glu Leu Glu Glu 1585 1590 1595 1600 GTT ATC CAG TAC AAA CTT GTC CCC GAG CGA CGA GAG ATC ATC CGC CAG ATC TGG 4833 4860 Val lie Gin Tyr Lys Leu Val Pro Glu Arg Arg Glu lie Arg Gin lie Trp 1605 1610 1615 TGG GAG AGA CTG CAG GGC TGC CAG CGT ATC GTA GAG GAC TGG CAG AAA ATC CTT 4887 4914 Trp Glu Arg Leu Gin Gly Cys Gin Argr lie Val Glu Asp Trp Gin Lys lie Leu 1620 1625 1630 1635 ATG GTG CGG TCC CTT GTG GTC AGC CCT CAT GAA GAC ATG AGA ACC TGG CTC AAG 4941 4968 MET Val Arg Ser Leu Val Val Pro Pro His Glu Asp MET Arg Thr Trp Leu Lys 1640 1645 1650 1655 TAT GCA AGC CTG TGC GGC AAG AGT GGC AGG CTG GCT CTT GOT CAT AAA ACT TTA 4995 5022 Tyr Ala Ser Leu Cys Gly Lys Ser Gly Arg Leu Ala Leu Ala His Lys Thr Leu 1660 1665 1670- / GTG TTG CTC CTG GGA GTT GAT CCG TCT CX3G CM CTT GAC CAT CCT CTG CCA ACA 5049 5076 Val Leu Leu Leu Gly Val Asp Pro Ser Arg Gin Leu Asp His Pro Leu Pro Thr 1675 1680 1685 169— > i fin in 1- ^ ϋ— I l_t I aw I m *-(please fill in the note f on page 5¾ and then fill out this page) Printed by the Central Consumers ’Coordination Department of the Ministry of Economic Affairs, Consumer Cooperatives it GTT CAC CCT CAG GTG ACC TAT GCC TAC ATG AAA AAC ATG TGG AAG AGT GCC CGC 5103 5130 Val His Pro Gin Val Thr Tyr Ala Tyr MET Lys Asn MET trp Lys Ser AXa Arg 1695 1700 1705 AAG ATC GAT GCC TTC CAG CAC ATG CAG CAT TTT GTC CAG ACC ATG CAG CAA CAG 5157 5184 Lys lie Aep Ala Phe Gin His MET Gin His Phe Val Gin Thr MET Gin Gin Gin 1710 1715 1720 1725 GCC CAG CAT GCC ATC GCT ACT GAG GAC CAG CAG CAT AAG CAG GAA CTG CAC AAG 5211 5238 Ala Gin His Ala He Ala Thr Glu Asp Gin Gin His Lys Gin Glu Leu His Lys 1730 1735 1740 1745 CTC ATG GCC CGA TGC TTC CTG AAA CTT GGA GAG TGG CAG CTG AAT CTA CAG GGC 5265 5292 Leu MET Ala Arg Cys Phe Leu Lys Leu Gly Glu Trp Gin Leu Asn Leu Gin Gly 1750 1755 1760 ATC GAG AGC ACA ATC CCC AAA GTG CTG CAG TAC TAC AGC GCC GCC ACA GAG 5319 5346 lie Asn Glu Ser Thr lie Pro Lys Val Leu Gin Tyr Tyr Ser Ala Ala Thr Glu 1765 1770 1775 1780 ) A4 specification (210 X 2W mm) 404851 A8 Βδ C8 D8 Patent application scope Printed by the Central Bureau of Standards of the Ministry of Economic Affairs Only Consumer Cooperatives CAC GAC CGC AGC TGG TAC AAG GCC TGG CAT GCG TGG GCA GTG ATG AAC TTC GAA 5373 5400 His Asp Arg Ser Trp Tyr Lys Ala Trp His Ala Trp Ala Val MET Asn Phe Glu 1785 1790 1795 GCT GTG CTA CAC TAC AAA CAT CAG AAC CAA GCC CGC GAT GAG AAG AAG AAA CTG 5427 5454 Ala Val Leu His Tyr Lys His Gin Asn Gin Ala Arg Asp Glu Lys Lys Lys Leu 1800 1805 1810 1815 CGT CAT GCC AGC GGG GCC AAC ATC ACC AAC GCC ACC ACT GCC GCC ACC ACG GCC 5481 5508 Arg His Ala Ser Gly Ala A sn lie Thr Asn Ala Thr Thr Ala Ala Thr Thr Ala χΘ20 1825 1830 1835 GCC ACT GCC ACC ACC ACT GCC AGC ACC GAG GGC AGC AAC AGT GAG AGC GAG GCC 5535 5562 Ala Thr Ala Thr Thr Thr Ala Ser Thr Glu Gly Ser Ash Glu Ser Glu Ala 1840 1845 1850 GAG AGC ACC GAG AAC AGC CCC ACC CCA TCG CCG CTG CAG AAG AAG GTC ACT GAG 5589 5616 Glu Ser Thr Glu Asn Ser Pro Thr Pro Ser Pro Leu Gin Lys Lys Val Thr Glu 1855 1860 1865 〆1870 GAT CTG TCC AAA ACC CTC CTG ATG TAC ACG OTG CCT GCC GTC CAG GGC TTC TTC 5643 5670 Asp Leu Ser Lys Thr Leu Leu MET Tyr Thr Val Pro Ala Vai Gin Gi.y Fhe Phe 1875 1880 1885 CGT TCC ATC TCC TTG TCA CGA GGC AAC AAC CTC CAG GAT ACA CTC AGA GTT CTC 5697 5724 Arg Ser lie Ser Leu Ser Arg Gly Asn Asn Leu Gin Asp Thr Leu'Arg Val Leu 1890 1895 1900 1905 ACC TTA TGG TTT GAT TAT GGT CAC TGG CCA GAT GTC AAT GAG GCC TTA GTG GAG 5751 5778 Thr Leu Trp Phe Asp Tyr Gly His Trp Pro Asp Val Asn Glu Ala Leu Val Glu 1910 1915 1920 1925 GGG GTG AAA GCC ATC CAG ATT GAT ACC TGG CTA CAG GTT ATA CCT CAG CT C ATT 5805 5832 Gly Val Lys Ala lie Gin lie Asp Thr Trp Leu Gin Val lie Pro Gin Leu lie 1930 1935 1940 GCA AGA ATT GAT ACG CCC AGA CCC TTG GTG GGA CGT CTC ATT CAC CAG CTT CTC 5859 5886 Ala Arg lie Asp Thr Pro Arg Pro Leu Val Gly Arg Leu lie His Gin Leu Leu 1945 1950 1955 1960 ACA GAC ATT GGT CGG TAC CAC CCC CAG GCC CTC ATC TAC CCA CTG ACA GTG GCT 5913 5940 Thr Asp lie Gly Arg Tyr His Pro Gin Ala Leu lie Tyr Pro Leu Thr Val Ala 1965 1970 1975 —f 〇 The paper scale is applicable to China National Standard (CNS) A4 specification (2! 0X297 mm) n II----11-I -I --- I- t I -III -I--In -HI (please read the note f on the back before filling this page) A8 B8404951 dI VI. Application for Patent Scope Central Government Bureau of the Ministry of Economic Affairs x Consumer Cooperatives Printing Policy TCX AAG TCT ACC ACG ACA GCC CGG CAC AAT GCA GCC AAC AAG ATT CTG AAG AAC 5967 5994 Ser Lys Ser Thr Thr Thr Ala Arg His Asn Ala Ala Asn Lys lie Leu Lys Asn 1980 1985 1990 1995 ATG TGT GAG CAC AGC AAC ACC CTG GTC CAG CAG GCC ATG ATG GTG AGO GAG GAG € 021 6046 MET Cys Glu H is Ser Asn Thr Leu Val Gin Gin Ala MET MET Val Ser Glu Glu 2000 2005 2010 2015 CTG ATC CGA GTG GCC ATC CTC TGG CAT GAG ATG TGG CAT GAA GGC CTG GAA GAG 6075 6102 Leu lie Arg Val Ala lie Leu Trp His Glu MET Trp His Glu Gly Leu Glu Glu 2020 2025 2030 GCA TCT CGT TTG TAC TTT GGG GAA AGG AAC GTG AAA GGC ATG TTT GAG GTG CTG 6129 6156 Ala Ser Arg Leu Tyr Phe Gly Glu Arg Asn Val Lys Gly MET Phe Glu Val Leu 2035 2040 2045 2050 GAG CCC TTG CAT GCT ATG ATG GAA CGG GGC CCC CAG ACT CTG AAG GAA ACA TCC 6183 6210 Glu Pro Leu His Ala MET MET Glu Arg Gly Pro Gin Thr Leu Lys Glu Thr Ser 2055 2060 2065 〆TTT AAT CAG GCC TAT GGT CX3A GAT ΤΓΑ ATG GAG GCC CM GAG TGG TGC AGG AAG 6237 6264 Phe Asn Gin Ala Tyr Giy Arg Asp Leu MET Giu Ala Gin Giu Trp Cys Arg 2070 2075 2080 2085 TAC ATG AAA TCA GGG AAT GTC AAG GAC TCC GAC CTC TAT TAT 6291 6318 Tyr MET Lys Ser Gly Asn Val Lys Asp I ^ eu Thr Gin Ala Trp Asp Leu Tyr Tyr 2090 2095 2100 2105 CAT GTG TTC CGA CGA ATC TCA AAG CAG CTG CCT CAG CTC A CA TCC TTA GAG CTG 6345 6372 His Val Phe Arg Arg lie Ser Lys Gin Leu Pro Gin Leu Thr Ser Leu Glu Leu 2110 2115 2120 CAA TAT GTT TCC CCA AAA CTT CTG ATG TGC CGG GAC CTT GAA TTG GCT GTG CCA 6399 6426 Gin Tyr Val Ser Pro Lys Leu Leu MET Cys Arg Asp Leu Glu lieu Ala Val Pro 2125 2130 2135 2140 GGA ACA TAT GAC CCC AAC CAG CCA ATC ATT CGC ATT CAG TCC ATA GCA CCG TCT 6453 6480 Gly Thr Tyr Asp Pro Asn Gin Pro He He Arg lie Gin Ser lie Ala Pro Ser 2145 2150 2155 TTG CAA GTC ATC ACA TCC AAG CAG AGG CCC CGG AAA TTG ACA CTT ATG GGC AGC € 507 6534 Leu Gin Val lie Thr Ser Lys Gin Arg Pro Arg Lys Leu Thr Leu MET Gly Ser 2160 2165 2170 2175 n 11 I 1 1 I ^^ 1 III 1 In 1 mn In--(please read the note f on the first page before filling in this note) Each paper size is in accordance with China B standards (CNS) A4 said Grid (2 丨 0 X 297 mm) A8 B8 C8 D8 ATG ATG 404S51 Patent Application Park AAC GGA CAT GAG TTT GTT TTC CTT CTA AAA GGC CAT GAA GAT CTG CGC CAG GAT 6561 6588 Asn Gly His Glu Phe Val Phe Leu Leu Lys Gly His Glu Asp Leu Arg Gin A sp 2180 2185 2190 2195 GAG CGT GTG ATG CAG CTC TTC GGC CTG GTT AAC ACC CTT CTG GCC AAT GAC CCA 6615 6642 Glu Arg Val MET Gin I ^ eu Phe Gly Leu Val Asn Thr I * eu Leu Ala Asn Asp Pro 2200 2205 2210 ACA TCT CTT CGG AAA AAC CTC AGO ATC CAG AGA TAC GCT GTC ATC CCT TTA TCG 6669 6696 Thr Ser Leu Arg Lys Asn Leu Ser He Gin Arg Tyr Ala Val lie Pro Leu Ser 2215 2220 2225 2230 ACC AAC TCG GGC CTC ATT GGC TGG GTT CCC CAC TGT GAC ACA CTG CAC GCC CTC 6723 6750 Thr Asn Ser Gly Leu lie Gly Trp Val Pro His Cys Asp Thr Leu His Ala Leu 2235 2240 2245 ATC CGG GAC TAC AGG GAG AAG AAG AAG ATC CTT CTC AAC ATC GAG CAT CGC ATC 6777 6804 lie Arg Asp Tyr Arg Glu Lys Lys Lys lie Leu Leu Asn lie Glu His Arg lie 2250 2255 2260 2265 'CCX3 GAC TAT GAC CAC TTG ACT CTG ATG CAG AAG'GTG GAS .6831 6858 MET Leu Arg MET Ala Pro Asp Tyr Asp His Leu Thr Leu MET Gin Lys Vai Giu 2270 2275 2280 2285 GTG TTT GAG CAT GCC GTC AAT AAT ACA GCT GGG GAC GAC CTG GCC AAG CTG CTG 6885, 6912 Val Phe Glu His Ala Val Asn A sn Thr Ala Gly Asp Asp Leu Too Li Lys Leu Leu 2290 2295 2300 TGG CTG AAA AGC CCC AGC TCC GAG GTG TGG TTT GAC CGA AGA ACC AAT TAT ACC 6939 6966 Trp Leu Lys Ser Pro Ser Ser Glu Val Trp Phe Asp Arg Arg Thr Asn Tyr Thr 2305 2310 2315 2320 CGT TCT TTA GCG GTC ATG TCA ATG GTT GGG TAT ATT TTA GGC CTG GGA GAT AGA 6993 7020 Arg Ser Leu Ala Val MET Ser MET Val Gly Tyr lie Leu Gly Leu Gly Asp Arg 2325 2330 2335 CAC CCA TCC AAC CTG ATG CTG GAC CGT CTG AGT GGG AAG ATC CTG CAC ATT GAC 7047 7074 His Pro Ser Asn Leu MET Leu Asp Arg Leu Ser Gly Lys lie Leu His lie Asp 2340 2345 2350 2355 TTT GGG GAC TGC TTT GAG GTT GCT ATG ACC CGA GAG AAG TTT CCA GAG AAG ATT 7101 7128 Phe Gly Asp Cys Phe Glu Val Ala MET Thr Arg Glu Lys Phe Pro Glu Lys He 2360 2365 2370 2375 This paper ruler Jt is applicable to China-Landmark (CNS) A4 specifications ( 210X297 public address) n «ii H In tn n in I. II i UK 、 ^^ (please read the" $ "on the back side before filling in this summer) Printed by the Consumer Affairs Cooperative of the Lost Gap Bureau in the Ministry of Economic Affairs 11 A8 B8 ^ 04951_dI Patent Application Scope Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs *. CCA TTT AGA CTA ACA AGA ATG TTG ACC AAT GOT ATG GAG GTT ACA GGC CTG GAT 7155 7182 Pro Phe Arg Leu Thr Arg MET Leu Thr Asn Ala MET Glu Val Thr Gly Leu Asp 2380 2385 2390 GGC AAC TAC AGA ATC ACA TGC CAC ACA GTG ATG GAG GTG CTG CGA GAG CAC AAG 7209 7236 Gly Asn Tyr Arg lie Thr Cys His Thr Val MET Glu Val Leu Arg Glu His hys 2395 2400 2405 2410 GAC AGT GTC ATG GCC GTG CTG GAA GCC TTT GTC TAX GAC CCC TTG CTG AAC TGG 7263 7290 Asp Ser Val MET Ala Val Leu Glu Ala Phe Val Tyr Asp Pro Leu Leu Asn Trp 2415 2420 2425 AGG CTG ATG GAC ACA AAT ACC AAA GGC AAC AAG CGA TCC CGA ACG AGG ACG GAT 7317 7344 Arg Leu MET Asp Thr Asn Thr Lys Gly Asn Lys Arg Ser Arg Thr Arg Thr Asp 2430 2435 2440 2445 TCC TAC TCT GCT GGC CAG TCA GTC GAA ATT TTG GAC GGT GTG GAA CTT GGA GAG 7371 Ser Tyr Ser Ala Gly Gin Ser Val Glu lie Leu Asp Gly Val Glu Leu Gly Glu 2450 2455 2460 2465 CQA GCC CAT AAG AAA ACQ_ GGG ACC ACA GTG CCA GAA TCT ATT CAT TCT T TC ATT-~ 7425 7452 Pro Ala His Lys Lys Thr Glv Thr Thr Val Pro Glu Ser lie His Ser Phe · He 2470 2475 2480 GGA GAC GGT TTG GTG AAA CCA GAG GCC CTA AAT AAG AAA GCT ATC CAG ATT ATT 7479 7506 Gly Asp Gly Leu Val Lys Pro Glu Ala Leu Asn Lys I * ys Ala lie Gin lie He 2485 2490 2495 2500 AAC AGG GTT CGA GAT AAG CTC ACT GGT CGG GAC TTC TCT CAT GAT GAC ACT TTG 7533 7560 Asn Arg Val Arg Λ $ ρ Lys Leu Thr Gly Arg Asp Phe Ser His Asp Asp Thr Leu 2505 2510 2515 GAT GTT CCA ACG CAA GTT GAG CTG CTC ATC AAA CAA GCG ACA TCC CAT GAA AAC 7587 7614 Asp Val Pro Thr Gin Val Glu Leu Leu lie Lys Gin Ala Thr Ser His Glu Asn 2520 2525 2530 2535 CTC TGC CAG TGC TAT ATT GGC TGG TAC CCT TTC TGG TAA 7641 Leu Cys Gin Cys Tyr lie Gly Trp Tyr Pro Phe Trp 2540 2545 2. —Substituted by the first scope of the patent application The amino acid sequence compiled by the acid sequence. -η- Applicable to Chinese paper size (CNS) A4 specifications (2 丨 0X297 mm) for each paper size I —————— It nnn I n I nn I i11-~ i (Fill in this page again) 404951 Application No ............. niolf Class Notice This amendment Gu A4 VBvU C4 Mold—Patent Specification 11) 4951 Chinese Rayberry Messin Effector Protein (86 years) (Amended in August) EFFECTOR PROTEINS OF RAPAMYCIH invented a new type of name. Inventor and creator of the name of the country. 1. MOLNAR-KIMBER, Katherine Lu> 2. CHEN YanqU 3. MAKAHISHI, Koji 4 FAILLI, Amedeo Arturo 5. CAGGIANO, Thomas Joseph 1. United States 2. People's Republic of China 3. Japan 4. Canada 5. United States 、 Home 1. Beautiful Garden, 19Θ38, 222 Granside, Dresden Road, New York, USA 2. New York, USA, 21, New York, “3EA, Di East Street, 427 3. The United States turns around 10021, New York ’s Redhead Road Apartment 56Θ 9J 4. Princeton, New Jersey Θ8550 Kessen Landinger No. 14 5. 350SS, Moresville, Stahan Road, Pennsylvania 19067, United States. Name (name), National Labor Department Central Ladder Bureau, Male Workers Consumer Cooperatives, Indian Nationality 1. Meipu Family Products Co., Ltd. ( Aierican Hose Products Corporation) 2. The Board of Trustees of Columbia University of New York (THE TRUSTEES OF COLIIKBIA UNIVERSITY IN THE CITY OF NEW YORK) 1. United States 2. US Line III. Applicant's Residence and Residence (Office) Name of Representative 1. New Zealand, USA 07940-0874 West Mandison 5 Gillard Farm 2. West 116th Street, Brocade Road, New York City, New York, USA 1. Egon E. Berg 2. Jack M. Granover Special (Jack M. Granowitz) Each paper uses the Chinese national standard rate (CNS > A4 threat (210X297)) according to the standard year ^ Amendment 40495J., Sa only before filling A7 Β7 V. Description of the invention (8) (please first (Please read the notes on the back and fill in this page again) -FKBP was isolated, Diels Adel adduct binds to FKBP12 and inhibits the PPIase activity of FOP1 2, but does not show detectable immunosuppressive activity, so it is not related to Repa In combination with GST-FKBP12, these two compounds were used in a similar separation step (ie, instead of rapastatin: GST-FKBP12) to produce a background concentration of 210kDa protein (without rapamectin) (Table 1). FK506, an immunosuppressive compound, has at least certain functions via the FK506-FKBP complex and calcium button binding mediators. * FK506 complexes with GST-FKBP in similar steps only produces 210 kDa protein at background concentrations (Table 1). Table 1 Comparison of rapamycin analog-FKBP12 complex and 210kDa protein binding Compound_ 210kDa LAF PPlaseiKi) RAPA + + + 6nM 0.12nM demethoxyrapamin (immunosuppressive analog) + + + 58nM 4.4 nil Diels Adele Adduct (Phenyl) Earth> 1000nM 12nM Diels Adele Adduct (Methyl) + 1000nM 12nM FK506 3Nra * 0.4nM None (FKBP) (* Mechanism of action different) The action of rapamycin, its analogues and other agents such as FK 5 06 suppresses the immune system. They perform this kind of action by binding to the respective hippoproteins. This binding inhibits the receptor protein's PPI a se activity However, inhibiting PPIase activity alone in combination with rapamycin or its analogs does not only cause immune suppression, it is indeed due to two rapamycin analogs (Diels Alder adduct in the table) adducts)) combined with FKBP subject to -25- This paper size is applicable to China National Standard (CNS) A4 specification (210X2 blade public director T Central Standards Bureau of the Ministry of Economy It encodes a part of human-derived peptone that can be bound to the GST-FKBP-rapamycin complex, and the sequence is as follows: AAG ATG CTT GGA ACC GGA CCT GCC GCC GCC ACC ACC GCT GCC ACC ACA TOT AGC 27 54 Met Leu Gly Thr Gly Pro Ala Ala Ala Thr Thr Ala Ala Thr Thr Ser Ser 15 10 15 AAT GTG AGC GTC CTG CAG CAG TTT GCC AGT GGC CTA AAG AGC CGG AAT GAG GAA 81 108 Asn Val Ser Val Leu Gin Gin Phe Ala Ser Gly Leu Lys Ser Arg Asn Glu Glu »20 25 30 35 ACC AGG GCC AAA GCC GCC AAG GAG CTC CAG CAC TAT GTC ACC ATG GAA CTC CGA 135 162 Thr Arg Ala Lys Ala Ala Lys Glu Leu Gin His Tyr Val Thr MET Glu Leu Arg 40 45 50 GAG ATG AGT CAA GAG GAG TCT ACT CGC TTC TAT GAC CAA CTG AAC CAT CAC ATT 189 216 Glu MET Ser Gin Glu Glu Ser Thr Arg Phe Tyr Asp Gin Leu Asn His His lie 55 60 65 70 TTT GAA TTG GTT TCC AGC TCA GAT GCC AA T GAG AGG AAA GGT GGC ATC TTG GCC 243 270 Phe Glu Leu Val Ser Ser Ser Asp Ala Asn Glu Arg Lys Gly Gly lie Leu Ala 75 80 85 ATA GCT AGC CTC ATA GGA GTG GAA GGT GGG AAT GCC ACC CGA ATTjGGC AGA. TTT 297 324 He Ala Ser Leulie Gly Val Glu Gly Gly Asn Ala Thr Arg He Gly Arg Phe 90 95 100 105 GCC AAC TAT CTT CGG AAC CTC CTC CCC TCC AAT GAC CCA GTT GTC ATG GAA ATG 351 378 Ala Asn Tyr Leu Arg Asn Leu Leu Pro Ser Asn Asp Pro Val Val MET Glu MET 110 115 120 125 GCA TCC AAG GCC ATT GGC CGT CTT GCC ATG GCA GGG GAC ACT TTT ACC GCT GAG 405 432 Ala Ser Lys Ala lie Gly Arg Leu Ala MET Ala Gly Asp Thr Phe Thr Ala Glu 130 135 140 TAC GTG GAA TTT GAG GTG AAG CGA GCC CTG GAA TGG CTG GGT GCT GAC CGC AAT 459 486 Tyr Val Glu Phe Glu Val Lys Arg Ala Leu Glu Trp Leu Gly Ala Asp Arg Asn 145 150 155 160 Sheet The scale is applicable to the rate of the middle and poor countries (CNS) A4 (210X297 mm) in ^ — n ^ in ^ i 1 ^ 1 im III— —I— —f ^ in ^ —-(Please read the note on the back first ^ • enter this page again)