TW202000918A - 超音波感應蛋白質以及利用超音波刺激細胞的方法 - Google Patents

超音波感應蛋白質以及利用超音波刺激細胞的方法 Download PDF

Info

Publication number
TW202000918A
TW202000918A TW107120094A TW107120094A TW202000918A TW 202000918 A TW202000918 A TW 202000918A TW 107120094 A TW107120094 A TW 107120094A TW 107120094 A TW107120094 A TW 107120094A TW 202000918 A TW202000918 A TW 202000918A
Authority
TW
Taiwan
Prior art keywords
leu
val
ala
gly
ile
Prior art date
Application number
TW107120094A
Other languages
English (en)
Other versions
TWI720319B (zh
Inventor
林玉俊
葉秩光
Original Assignee
國立清華大學
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 國立清華大學 filed Critical 國立清華大學
Priority to TW107120094A priority Critical patent/TWI720319B/zh
Priority to US16/290,214 priority patent/US20190374763A1/en
Publication of TW202000918A publication Critical patent/TW202000918A/zh
Application granted granted Critical
Publication of TWI720319B publication Critical patent/TWI720319B/zh

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M37/00—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin
    • A61M37/0092—Other apparatus for introducing media into the body; Percutany, i.e. introducing medicines into the body by diffusion through the skin using ultrasonic, sonic or infrasonic vibrations, e.g. phonophoresis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0002—Galenical forms characterised by the drug release technique; Application systems commanded by energy
    • A61K9/0009—Galenical forms characterised by the drug release technique; Application systems commanded by energy involving or responsive to electricity, magnetism or acoustic waves; Galenical aspects of sonophoresis, iontophoresis, electroporation or electroosmosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N7/00—Ultrasound therapy
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N7/00—Ultrasound therapy
    • A61N2007/0004—Applications of ultrasound therapy
    • A61N2007/0021—Neural system treatment
    • A61N2007/0026—Stimulation of nerve tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N7/00—Ultrasound therapy
    • A61N2007/0056—Beam shaping elements
    • A61N2007/006—Lenses

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Biomedical Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Toxicology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Radiology & Medical Imaging (AREA)
  • Molecular Biology (AREA)
  • Dermatology (AREA)
  • Medical Informatics (AREA)
  • Anesthesiology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Hematology (AREA)
  • Cell Biology (AREA)
  • Immunology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

本發明提供了一種超音波感應蛋白質，其係具有無法感受超音波的哺乳動物耳蝸外毛細胞的馬達蛋白胺基酸序列第308個位置的胺基酸被另一個胺基酸取代的變異、以及選擇性地具有第7個位置的胺基酸被另一個胺基酸取代的變異。

Description

超音波感應蛋白質以及利用超音波刺激細胞的方法
本發明係有關於一種超音波感應蛋白質以及利用超音波刺激細胞的方法，尤指一種無法感受超音波的哺乳動物耳蝸外毛細胞的變異型馬達蛋白質以及利用超音波刺激表現該超音波蛋白質的細胞。
在自然界中，各種生物體能夠感測到不同的環境因素並對應的做出反應，例如可感測到光、熱、化學物質、或磁場。而生物體感受各種刺激的因素在於細胞內的特殊蛋白質，各種特殊蛋白質受到不同環境因素的刺激，可進一步影響細胞的生理活性，因此，科學家們利用這種反應機制而將各類的特殊蛋白質嵌入人體細胞內，以賦予細胞感受如光、熱、化學物質、或磁場刺激的能力，如此一來，即可利用不同的刺激來控制特定細胞的生理活性，從而可達到不同的醫療目的。
然而，目前雖已發展了可感受到光、化學物質、或磁場的蛋白質，若欲應用在醫療目的上，則遭受到很大的限制。舉例而言，若將可感應光的蛋白質嵌入人體特定組織的細胞內時，光的穿透能力不佳，無法直接照射到體內深處的細胞，而光源需以侵入式的方法進行照射以刺激特定蛋白質，進而控制細胞的生理活性，然而，侵入式的光源仍需要藉由外科手術而進入體內進行照射，造成患者生理上更大的負擔。而若將可感受化學物質或磁場刺激的蛋白質嵌入特定組織的細胞內時，則會有難以定位細胞使得刺激效果緩慢或刺激效果不佳的問題。
因此，目前急需一種非侵入式，且可準確地於生物體內定位並給予細胞刺激的方法，以達到可準確控制特定細胞的生理活性的目的。
為了達到上述之目的，本發明提供了一種改良自然界中與高頻聽覺有關的蛋白質，其具有感受特定頻率超音波刺激的能力，若將此蛋白質放入人類細胞中或使細胞可表現該蛋白質，即可賦予該細胞感受超音波刺激的特性，藉此可使用醫用的聚焦式超音波，以非侵入性的方法刺激特定細胞，影響其生理活性，並達到醫療的目的。
本發明提供了一種超音波感應蛋白質，其係具有無法感受超音波的哺乳動物耳蝸外毛細胞的馬達蛋白胺基酸序列第308個位置的天門冬醯胺(Asparagine)被絲氨酸(Serine)取代的變異、以及選擇性地具有第7個位置的天門冬醯胺(Asparagine)被蘇胺酸(Threonine)取代的變異。
於本發明一較佳實施態樣中，該超音波感應蛋白質係具有無法感受超音波的哺乳動物耳蝸外毛細胞的馬達蛋白胺基雖序列第308個位置的天門冬醯胺被絲氨酸取代的變異、以及第7個位置的天門冬醯胺被蘇胺酸取代的變異。
於本發明一實施態樣中，該無法感受超音波的哺乳動物的耳蝸外毛細胞的馬達蛋白係人類、小鼠、狐蝠、小鬚鯨、長舌果蝠、或犬蝠的耳蝸外毛細胞的馬達蛋白。其中，人類的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 1；小鼠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 2；狐蝠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 3；小鬚鯨的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 4；長舌果蝠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 5；以及，棕果蝠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 6。
於本發明中，人類的耳蝸外毛細胞的N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO: 7；人類的耳蝸外毛細胞的N7T、N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO: 8；小鼠的耳蝸外毛細胞的N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO: 9; 小鼠的耳蝸外毛細胞的N7T、N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO:10。
本發明更提供了一種刺激細胞的方法，包括利用一超音波照射表現一超音波感應蛋白質的一細胞，其中，該超音波感應蛋白質具有無法感受超音波的哺乳動物的耳蝸外毛細胞的馬達蛋白胺基酸序列第308個位置的天門冬醯胺(Asparagine)被絲氨酸(Serine)取代的變異(N308S)、以及選擇性地具有第7個位置的天門冬醯胺(Asparagine)被蘇胺酸(Threonine)取代的變異(N7T)。
於一較佳實施態樣中，該超音波感應蛋白質具有無法感受超音波的哺乳動物的耳蝸外毛細胞的馬達蛋白胺基酸序列第308個位置的天門冬醯胺被絲氨酸取代的變異、以及選擇性的具有第7個位置的天門冬醯胺被蘇胺酸取代的變異。
於本發明之一實施態樣中，該超音波係一對焦式超音波。此外，該超音波發射的頻率較佳為0.35~0.65 MHz；聲壓為0.1~1.0 MPa；其中又以頻率為0.5 MHz；聲壓為0.5MPa為最佳。
於本發明一實施態樣中，該細胞經該超音波照射後，被誘導鈣離子流入該細胞內。
於本發明一實施態樣中，該細胞為神經細胞、免疫細胞、胰島細胞、上皮細胞、血液細胞、肌肉細胞、或幹細胞等。
本發明所提供之超音波感應蛋白質可感應超音波的刺激，而當該超音波感應蛋白質嵌入細胞、或者表現該超音波感應蛋白質的細胞，經超音波照射可精準地控制其基因表現、控制神經活性或免疫反應等，並可發展各種不同的嶄新系統以利用超音波精準控制細胞的生理活性而達到各種醫療目的，對於醫療方法實為相當大的突破。
[ 細胞 培養 ]
為了找尋能感受超音波刺激的蛋白質，本發明比對了可感受超音波的哺乳動物聽覺系統內的耳蝸外毛細胞的馬達蛋白(Prestin)序列，以及無法感受超音波的哺乳動物聽覺系統內的耳蝸外毛細胞的馬達蛋白序列，發現在可感受超音波哺乳動物的Prestin蛋白質序列中第7個及第308個胺基酸分別為蘇氨酸(Threonine)及絲氨酸(Serine)，而無法感受超音波的哺乳動物的Prestin蛋白質序列的第7個及第308個胺基酸多為天門冬醯胺(Asparagine)，故本發明探討了將這些不同的氨基酸置換是否能在感受超音波上扮演重要角色。因此，本實施例將N7T及N308S點突變放入無法感受超音波的老鼠的Prestin蛋白質中，並測試其對於超音波刺激的反應。
293T細胞係使用包含DMEM(Dulbecco’s modified Eagle’s medium, Gibco)培養基、10%的胎牛血清(Fetal bovine serum)、5 U/mL的盤尼西林(Penicillin)、以及50 μg/mL的鏈黴素(streptomycin)的細胞培養液進行培養。
接著，293T細胞係利用轉染試劑LT-1(Mirus)分別將Venus-mPrestin(野型) (SEQ ID NO: 11)、Venus-mPrestin(N7T) (SEQ ID NO:12)、Venus-mPrestin(N308S) (SEQ ID NO: 13)、Venus-mPrestin(N7T, N308S) (SEQ ID NO: 14)的DNA片段轉染至293T細胞，以獲得表現Venus-mPrestin (野型)、Venus-mPrestin (N7T)、Venus-mPrestin (N308S)、以及Venus-mPrestin (N7T, N308S)的293T細胞。
[ 表現 Pre stin 細胞對於不同頻率的超音波的反應 ]
鈣離子通道打開與否常被作為細胞對於超音波等力學刺激是否有反應的指標。因此，在本實驗中，係利用容易滲透且不引起熱反應或造成組織傷害的低頻的超音波(＜3.5 MHz， 0.5 MPa，2000個週期)作為刺激來源，刺激表現红色鈣指示蛋白(R-GECO)以及表現黃色螢光蛋白(Venus)細胞作為控制組，以及刺激分別表現R-GECO與Venus-mPrestin (野型)、Venus-mPrestin (N7T)、Venus-mPrestin (N308S)、以及Venus-mPrestin (N7T, N308S)的細胞。
圖1的實驗利用基因轉殖將293T細胞分別表現Venus黃色螢光蛋白(控制組)及表現Venus-mPrestin (N7T, N308S)，此兩組細胞同時更表現红色鈣指示蛋白(R-GECO)。超音波刺激(0.5 MHz，0.5 MPa，2000個週期，3秒鐘)有效地刺激Venus-mPrestin (N7T, N308S)細胞的鈣離子反應，而使該細胞紅色鈣指示蛋白亮度增加，而同樣的超音波照射無法有效地刺激控制組細胞的鈣離子反應。
請參照圖1所示的細胞成像圖，其中綠螢光顯示控制組Venus蛋白質或Venus-mPrestin(N7T, N308S)蛋白質、青色螢光顯示細胞內的鈣指示蛋白質位置，而紅色螢光係顯示鈣離子的位置，故由圖1可見，0.5 MHz的超音波激發了表現Venus-mPrestin (N7T, N308S)的細胞的鈣離子反應，使得大量鈣離子流入細胞內，然而，作為控制組的細胞並未被超音波激發其鈣離子反應。
293T細胞經過基因轉殖後表現Venus、Venus-mPrestin (野型)、Venus-mPrestin (N7T)、Venus-mPrestin (N308S)、或Venus-mPrestin (N7T, N308S)，各組細胞還會同時表現紅色鈣離子指示蛋白，經過不同頻率的超音波刺激(80 kHz~3.5 MHz，0.5 MPa，2000個週期，3秒鐘)，計算有鈣離子反應的細胞比例，並將比例除以控制組的比例而統整表格如圖2。
請參照圖2，其係各組細胞中，被激發鈣離子反應的機率的倍數，該倍數係以控制組(Venus)做標準化計算。由圖2的結果可得知，經0.5 MHz超音波刺激的表現Venus-mPrestin (N308S)以及表現Venus-mPrestin (N7T, N308S)的細胞有明顯的鈣離子反應，而其中又以表現Venus-mPrestin (N7T, N308S)的細胞的鈣離子反應為控制組的約11倍，且有統計上的差異(p＜0.05)。
由以上結果可得知，表現mPrestin(N7T, N308S)的細胞對於0.5 MHz的超音波有高度的敏感性，且對於超音波產生反應的百分比為控制組細胞的11倍以上，因此可得知mPrestin(N7T, N308S)賦予轉染的哺乳動物細胞感應0.5 MHz的超音波的特性。
[ 非侵入式刺激表現 Prestin 細胞]
本說明書中所有涉及動物實驗的步驟皆受到國立清華大學動物委員會的批准而進行(IACUC號：NTHU10459)。
本實驗主要係將表現Venus-mPrestin(N7T, N308S)的DNA片段轉染至小鼠腦中的神經細胞中，使得小鼠神經細胞可表現Venus-mPrestin(N7T, N308S)蛋白質，再利用超音波刺激表現有Venus-mPrestin(N7T, N308S)蛋白質的神經細胞，觀察是否可利用超音波於不開腦的情況下激發腦內神經細胞的活性。本實驗的詳細的實驗方法如下。
將莫耳比為31.5：3.9：1.8的1,2-二棕櫚醯基-sn-甘油-3-磷酸膽鹼(1,2-dipalmitoyl-sn-glycero-3-phosphocholine; DPPC, Avanti Polar Lipids, AL, USA)、1,2-二棕櫚醯-3-三甲基銨-丙烷(1,2-dipalmitoyl-3-trimethylammonium-propane; DPTAP, Avanti Polar Lipids)、以及 1,2-二硬脂醯-sn-甘油-3-磷酸乙醇胺-N-[羧基(聚乙二醇)-2000 (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[carboxy(polyethyleneglycol)-2000]；DSPE-PEG 2000, Avanti Polar Lipids)溶於氯仿中，並乾燥24小時以上， 以形成乾燥的脂質膜。接著將該脂質膜與甘油-磷酸鹽緩衝生理實驗水(PBS)(5 μL/mL)以及全氟丙烷(C3 F8 )氣體於一攪拌器中攪拌45秒以形成微氣泡。未反應的脂質係利用離心方法(6000 rpm，兩分鐘)與微氣泡分離。DPTAP的陽離子特性可使得微氣泡藉由靜電作用而貼附於細胞質體(plasmid)上。
為了得到載有Venus-Prestin的DNA片段的微氣泡，5 μg的Venus-Prestin的DNA片段與50 μL濃度為2×108 個/μL的微氣泡混合液混合30分鐘後，再藉由離心方法(6000 rpm，2分鐘)分離未載有Venus-Prestin的DNA片段的微氣泡以及載有Venus-Prestin的DNA片段的微氣泡。成功結合DNA片段的微氣泡可透過碘化丙啶染色於顯微鏡觀察下成像，而DNA片段與微氣泡的結合效率可利用分光光度計並透過以下公式來評估：
Figure 02_image001
由以上實驗結果可得知，DNA結合效率為24.5±1.6%，實際承載於微氣泡之的DNA量為1.2±0.1 μg。
另外，載有Venus DNA片段的微氣泡同樣係藉由上述方法而製備作為對照組。
進行以下細胞轉染實驗前，使用異氟醚(isoflurane)(劑量：1%；流速：1 L/分鐘)以及純氧以麻醉9隻6-10週齡的雄性C57BL/6JNarl小鼠。
小鼠體內的基因轉染係利用1-MHz的聚焦式超音波換能器(半徑 = 38 mm，焦距 = 60 mm；V302, Panametrics, Waltham, MA, USA)以及上述所製備的載有DNA片段的微氣泡而進行，其詳細方法如下。
首先，將6隻小鼠隨機分為兩組：一組為注射載有Venus-Prestin DNA片段的微氣泡作為實驗組，另一組注射載有Venus DNA片段的微氣泡作為對照組。以上兩組中載有DNA片段的微氣泡係藉由眼眶後注射而注入小鼠體內，20秒後，利用聲壓為0.5 MPa，週期為10000，脈衝重複頻率為5 Hz的聚焦式超音波換能器經顱照射小鼠的左腦的兩個處理點，使得微氣泡攜帶的DNA片段可穿過血腦屏障並進入細胞以進行基因轉染。
基因轉染48小時後即利用0.5-MHz的聚焦式超音波換能器(V389，Panametrics)照射以進行體內的神經調節，其週期為2000、脈衝重複頻率為1 Hz，一個處理點照射3秒的超音波。另外，三隻未接受基因轉殖的正常小鼠亦接受0.5-MHz的超音波照射作為控制組。
經上述0.5-MHz的超音波照射後，犧牲小鼠並取出腦部，再製作15 μm的切片，該些切片係於-20°C的甲醇中固定20分鐘，並利用含有5%山羊血清以及1%牛血清白蛋白的PBS溶液浸泡該些切片以阻斷內源性蛋白質。接著，該些切片於利用抗體稀釋液稀釋1000倍的初代兔子抗-c-Fos抗體中浸置過夜，再與利用抗體稀釋液稀釋200倍的Dylight 594共軛的抗兔二抗中浸置1小時後，利用PBS清洗數次，細胞核係利用DAPI標示。最後，將載玻片蓋上螢光固定介質並於-20°C的黑暗中平放保存，最後使用光學顯微鏡進行免疫組織化學染色的觀察，其結果如圖3所示。由圖3可得知，通過Venus螢光蛋白的綠螢光表達證實了Venus-Prestin和Vneus的成功轉染，而c-fos抗體染色的紅螢光則顯示經超音波照射後被活化的細胞。
經上述實驗證實，本發明所提供的超音波刺激細胞的方法可將超音波感應蛋白嵌入細胞後，於體外照射超音波以控制表現超音波感應蛋白質的細胞的生理活性。
圖1為本發明實施例中，控制組及表現Venus-mPrestin (N7T, N308S)經超音波刺激後之細胞螢光影像圖。 圖2為本發明實施例中，各組細胞被激發鈣離子反應的機率的倍數數據圖。 圖3為本發明實施例中，非侵入式刺激表現Prestin的細胞的光學顯微鏡進行免疫組織化學染色的細胞螢光影像圖。
＜110＞ 國立清華大學 ＜120＞ 超音波感應蛋白質以及利用超音波刺激細胞的方法 ＜130＞ NTHU-I25 ＜160＞ 14 ＜170＞ PatentIn version 3.5 ＜210＞ 1 ＜211＞ 712 ＜212＞ PRT ＜213＞ Homo sapiens ＜400＞ 1 Met Asp His Ala Glu Glu Asn Glu Ile Leu Ala Ala Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Thr Lys Asp Lys Val Pro Asp Ser Ile Ala Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Ile Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Leu Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Asn Leu Lys Glu Ser Tyr Asn Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Leu Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Leu Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 435 440 445 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 450 455 460 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 465 470 475 480 Lys Leu Pro Glu Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 485 490 495 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 500 505 510 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Asn Ala Leu Lys Arg Lys Thr 515 520 525 Gly Val Asn Pro Ala Val Ile Met Gly Ala Arg Arg Lys Ala Met Arg 530 535 540 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Met Ala Asn Ala Thr Val 545 550 555 560 Val Lys Ala Asp Ala Glu Val Asp Gly Glu Asp Ala Thr Lys Pro Glu 565 570 575 Glu Glu Asp Gly Glu Val Lys Tyr Pro Pro Ile Val Ile Lys Ser Thr 580 585 590 Phe Pro Glu Glu Met Gln Arg Phe Met Pro Pro Gly Asp Asn Val His 595 600 605 Thr Val Ile Leu Asp Phe Thr Gln Val Asn Phe Ile Asp Ser Val Gly 610 615 620 Val Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile 625 630 635 640 Tyr Val Tyr Leu Ala Gly Cys Ser Ala Gln Val Val Asn Asp Leu Thr 645 650 655 Arg Asn Arg Phe Phe Glu Asn Pro Ala Leu Trp Glu Leu Leu Phe His 660 665 670 Ser Ile His Asp Ala Val Leu Gly Ser Gln Leu Arg Glu Ala Leu Ala 675 680 685 Glu Gln Glu Ala Ser Ala Pro Pro Ser Gln Glu Asp Leu Glu Pro Asn 690 695 700 Ala Thr Pro Ala Thr Pro Glu Ala 705 710 ＜210＞ 2 ＜211＞ 744 ＜212＞ PRT ＜213＞ Mus musculus ＜400＞ 2 Met Asp His Ala Glu Glu Asn Glu Ile Pro Ala Glu Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Val Lys Asp Lys Val Thr Glu Ser Ile Gly Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Val Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Phe Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Asn Leu His Glu Ser Tyr Ser Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Ile Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Leu Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Ala Val Leu Ser Ala Ile Val Ile Val Asn Leu 435 440 445 Lys Gly Met Phe Met Gln Phe Ser Asp Leu Pro Phe Phe Trp Arg Thr 450 455 460 Ser Lys Ile Glu Leu Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 465 470 475 480 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 485 490 495 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 500 505 510 Gln Leu Pro Asp Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 515 520 525 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 530 535 540 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Ser Ala Leu Lys Arg Lys Thr 545 550 555 560 Gly Val Asn Pro Ala Leu Ile Met Gly Ala Arg Arg Lys Ala Met Arg 565 570 575 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Val Ala Asn Ala Thr Val 580 585 590 Val Lys Val Asp Ala Glu Val Asp Gly Glu Asn Ala Thr Lys Pro Glu 595 600 605 Glu Glu Asp Asp Glu Val Lys Phe Pro Pro Ile Val Ile Lys Thr Thr 610 615 620 Phe Pro Glu Glu Leu Gln Arg Phe Leu Pro Gln Gly Glu Asn Val His 625 630 635 640 Thr Val Ile Leu Asp Phe Thr Gln Val Asn Phe Val Asp Ser Val Gly 645 650 655 Val Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile 660 665 670 Tyr Val Tyr Leu Ala Gly Cys Ser Pro Gln Val Val Asn Asp Leu Thr 675 680 685 Arg Asn Asn Phe Phe Glu Asn Pro Ala Leu Lys Glu Leu Leu Phe His 690 695 700 Ser Ile His Asp Ala Val Leu Gly Ser Gln Val Arg Glu Ala Met Ala 705 710 715 720 Glu Gln Glu Ala Thr Ala Ser Leu Pro Gln Glu Asp Met Glu Pro Asn 725 730 735 Ala Thr Pro Thr Thr Pro Glu Ala 740 ＜210＞ 3 ＜211＞ 740 ＜212＞ PRT ＜213＞ Pteropus vampyrus ＜400＞ 3 Met Asp His Ala Glu Glu Asn Glu Ile Leu Ala Ala Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Lys Lys Asp Lys Ile Pro Asp Ser Ile Gly Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Val Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Phe Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Asn Leu His Glu Ser Tyr Asn Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Leu Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Met Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Ala Val Leu Ser Ala Ile Val Ile Val Asn Leu 435 440 445 Lys Gly Met Phe Met Gln Phe Ser Asp Leu Pro Phe Phe Trp Arg Thr 450 455 460 Ser Lys Ile Glu Leu Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 465 470 475 480 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 485 490 495 Met Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 500 505 510 Gln Leu Pro Asp Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 515 520 525 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 530 535 540 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Asn Ala Leu Lys Arg Lys Thr 545 550 555 560 Gly Val Asn Pro Ala Leu Ile Met Gly Ala Arg Arg Lys Ala Met Arg 565 570 575 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Met Ala Asn Ala Ala Val 580 585 590 Val Lys Ala Asp Ala Glu Val Asp Gly Glu Asp Gly Thr Lys Pro Glu 595 600 605 Glu Glu Asp Glu Val Lys Tyr Pro Pro Ile Val Ile Lys Asn Thr Phe 610 615 620 Pro Glu Glu Leu Gln Arg Phe Met Pro Pro Gly Asp Asn Ile His Thr 625 630 635 640 Val Ile Leu Asp Phe Thr Gln Val Asn Phe Ile Asp Ser Val Gly Val 645 650 655 Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile Tyr 660 665 670 Val Tyr Leu Ala Gly Cys Ser Ala Gln Val Val Asn Asp Leu Thr Gln 675 680 685 Asn Gln Phe Phe Glu Asn Pro Ala Leu Arg Glu Leu Leu Phe His Ser 690 695 700 Ile His Asp Ala Val Leu Gly Ser His Val Arg Glu Ala Leu Ala Glu 705 710 715 720 Gln Glu Ala Ser Ala Pro Ala Thr Gln Glu Asp Met Glu Pro Asn Ala 725 730 735 Thr Pro Glu Ala 740 ＜210＞ 4 ＜211＞ 741 ＜212＞ PRT ＜213＞ Balaenoptera acutorostrata ＜400＞ 4 Met Asp His Ala Glu Glu Asn Glu Ile Leu Ala Ala Ala Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Leu Val Leu Gln Glu Arg Leu 20 25 30 His Lys Lys Asp Lys Ile Ser Asp Ser Ile Gly Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Arg Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Val Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Phe Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Met Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Asn Leu Asn Glu Ser Tyr Asn Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Leu Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Leu Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Ala Val Leu Ser Ala Ile Val Ile Val Asn Leu 435 440 445 Lys Gly Met Phe Met Gln Phe Ser Asp Leu Pro Phe Phe Trp Arg Thr 450 455 460 Ser Lys Ile Glu Leu Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 465 470 475 480 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 485 490 495 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 500 505 510 Gln Leu Pro Asp Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 515 520 525 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 530 535 540 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Ser Ala Leu Lys Arg Lys Thr 545 550 555 560 Gly Val Asn Pro Ala Leu Ile Met Gly Ala Arg Arg Lys Ala Met Lys 565 570 575 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Met Ala Asn Ala Thr Ile 580 585 590 Val Lys Val Asp Ala Glu Val Asp Gly Glu Asp Gly Thr Lys Pro Glu 595 600 605 Glu Glu Glu Asn Glu Ile Lys Tyr Pro Pro Val Val Ile Lys Ser Thr 610 615 620 Leu Pro Glu Glu Leu Gln Arg Phe Met Pro Pro Gly Asp Asn Val His 625 630 635 640 Thr Ile Ile Leu Asp Phe Thr Gln Val Asn Phe Ile Asp Ser Val Gly 645 650 655 Val Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile 660 665 670 Tyr Val Tyr Leu Ala Gly Cys Ser Ala Gln Val Val Asn Asp Leu Ser 675 680 685 Arg Asn Gln Phe Phe Glu Asn Pro Ala Leu Leu Glu Leu Leu Phe His 690 695 700 Ser Ile His Asp Ala Val Leu Gly Ser Gln Val Arg Glu Ala Leu Ala 705 710 715 720 Glu Gln Glu Ala Thr Ala Ala Pro Pro Gln Glu Asp Ser Glu Pro Asn 725 730 735 Ala Thr Pro Glu Ala 740 ＜210＞ 5 ＜211＞ 740 ＜212＞ PRT ＜213＞ Eonycteris spelaea ＜400＞ 5 Met Asp His Ala Glu Glu Asn Glu Ile Leu Ala Ala Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Lys Lys Asp Lys Ile Pro Asp Ser Ile Gly Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Val Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Phe Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Asn Leu His Glu Ser Tyr Asn Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Leu Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Met Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Ala Val Leu Ser Ala Ile Val Ile Val Asn Leu 435 440 445 Lys Gly Met Phe Met Gln Phe Ser Asp Leu Pro Phe Phe Trp Arg Thr 450 455 460 Ser Lys Ile Glu Leu Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 465 470 475 480 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 485 490 495 Met Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 500 505 510 Gln Leu Pro Asp Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 515 520 525 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 530 535 540 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Asn Ala Leu Lys Arg Lys Thr 545 550 555 560 Gly Val Asn Pro Ala Leu Ile Met Gly Ala Arg Arg Lys Ala Met Arg 565 570 575 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Met Ala Asn Ala Ala Val 580 585 590 Ile Lys Ala Asp Ala Glu Val Asp Gly Asp Asp Gly Thr Lys Pro Glu 595 600 605 Glu Glu Asp Glu Val Lys Tyr Pro Pro Ile Val Ile Lys Asn Thr Phe 610 615 620 Pro Glu Glu Leu Gln Arg Phe Met Pro Pro Gly Asp Asn Val His Thr 625 630 635 640 Val Ile Leu Asp Phe Thr Gln Val Asn Phe Ile Asp Ser Val Gly Val 645 650 655 Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile Tyr 660 665 670 Val Tyr Leu Ala Gly Cys Ser Ala Gln Val Val Asn Asp Leu Thr Gln 675 680 685 Asn Gln Phe Phe Glu Asn Pro Ala Leu Arg Glu Leu Leu Phe His Ser 690 695 700 Ile His Asp Ala Val Leu Gly Ser His Val Arg Glu Ala Leu Ala Glu 705 710 715 720 Gln Glu Ala Ser Ala Pro Thr Thr Gln Glu Asp Leu Glu Pro Asn Ala 725 730 735 Thr Pro Glu Ala 740 ＜210＞ 6 ＜211＞ 740 ＜212＞ PRT ＜213＞ Rousettus leschenaultia ＜400＞ 6 Met Asp His Ala Glu Glu Asn Glu Ile Leu Ala Ala Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Lys Lys Asp Lys Ile Pro Asp Ser Ile Gly Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Val Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Phe Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Asn Leu His Glu Ser Tyr Asn Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Leu Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Met Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Ala Val Leu Ser Ala Ile Val Ile Val Asn Leu 435 440 445 Lys Gly Met Phe Met Gln Phe Ser Asp Leu Pro Phe Phe Trp Arg Thr 450 455 460 Ser Lys Ile Glu Leu Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 465 470 475 480 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 485 490 495 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 500 505 510 Gln Leu Pro Asp Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 515 520 525 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 530 535 540 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Asn Ala Leu Lys Arg Lys Thr 545 550 555 560 Gly Val Asn Pro Ala Phe Ile Met Gly Ala Arg Arg Lys Ala Met Arg 565 570 575 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Met Ala Asn Ala Ala Val 580 585 590 Ile Lys Ala Asp Ala Glu Val Asp Gly Glu Asp Gly Thr Lys Pro Glu 595 600 605 Glu Glu Asp Glu Val Lys Tyr Pro Pro Ile Val Ile Lys Asn Thr Phe 610 615 620 Pro Glu Glu Leu Gln Arg Phe Met Pro Pro Gly Asp Asn Ile His Thr 625 630 635 640 Val Ile Leu Asp Phe Thr Gln Val Asn Phe Ile Asp Ser Val Gly Val 645 650 655 Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile Tyr 660 665 670 Val Tyr Leu Ala Gly Cys Ser Ala Gln Val Val Asn Asp Leu Thr Gln 675 680 685 Asn Gln Phe Phe Glu Asn Pro Ala Leu Arg Glu Leu Leu Phe His Ser 690 695 700 Ile His Asp Ala Val Leu Gly Ser His Val Arg Glu Ala Leu Ala Glu 705 710 715 720 Gln Glu Ala Ser Ala Pro Thr Thr Gln Glu Asp Leu Glu Pro Asn Ala 725 730 735 Thr Pro Glu Ala 740 ＜210＞ 7 ＜211＞ 712 ＜212＞ PRT ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 7 Met Asp His Ala Glu Glu Asn Glu Ile Leu Ala Ala Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Thr Lys Asp Lys Val Pro Asp Ser Ile Ala Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Ile Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Leu Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Ser Leu Lys Glu Ser Tyr Asn Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Leu Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Leu Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 435 440 445 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 450 455 460 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 465 470 475 480 Lys Leu Pro Glu Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 485 490 495 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 500 505 510 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Asn Ala Leu Lys Arg Lys Thr 515 520 525 Gly Val Asn Pro Ala Val Ile Met Gly Ala Arg Arg Lys Ala Met Arg 530 535 540 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Met Ala Asn Ala Thr Val 545 550 555 560 Val Lys Ala Asp Ala Glu Val Asp Gly Glu Asp Ala Thr Lys Pro Glu 565 570 575 Glu Glu Asp Gly Glu Val Lys Tyr Pro Pro Ile Val Ile Lys Ser Thr 580 585 590 Phe Pro Glu Glu Met Gln Arg Phe Met Pro Pro Gly Asp Asn Val His 595 600 605 Thr Val Ile Leu Asp Phe Thr Gln Val Asn Phe Ile Asp Ser Val Gly 610 615 620 Val Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile 625 630 635 640 Tyr Val Tyr Leu Ala Gly Cys Ser Ala Gln Val Val Asn Asp Leu Thr 645 650 655 Arg Asn Arg Phe Phe Glu Asn Pro Ala Leu Trp Glu Leu Leu Phe His 660 665 670 Ser Ile His Asp Ala Val Leu Gly Ser Gln Leu Arg Glu Ala Leu Ala 675 680 685 Glu Gln Glu Ala Ser Ala Pro Pro Ser Gln Glu Asp Leu Glu Pro Asn 690 695 700 Ala Thr Pro Ala Thr Pro Glu Ala 705 710 ＜210＞ 8 ＜211＞ 712 ＜212＞ PRT ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 8 Met Asp His Ala Glu Glu Thr Glu Ile Leu Ala Ala Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Thr Lys Asp Lys Val Pro Asp Ser Ile Ala Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Ile Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Leu Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Ser Leu Lys Glu Ser Tyr Asn Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Leu Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Leu Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 435 440 445 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 450 455 460 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 465 470 475 480 Lys Leu Pro Glu Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 485 490 495 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 500 505 510 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Asn Ala Leu Lys Arg Lys Thr 515 520 525 Gly Val Asn Pro Ala Val Ile Met Gly Ala Arg Arg Lys Ala Met Arg 530 535 540 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Met Ala Asn Ala Thr Val 545 550 555 560 Val Lys Ala Asp Ala Glu Val Asp Gly Glu Asp Ala Thr Lys Pro Glu 565 570 575 Glu Glu Asp Gly Glu Val Lys Tyr Pro Pro Ile Val Ile Lys Ser Thr 580 585 590 Phe Pro Glu Glu Met Gln Arg Phe Met Pro Pro Gly Asp Asn Val His 595 600 605 Thr Val Ile Leu Asp Phe Thr Gln Val Asn Phe Ile Asp Ser Val Gly 610 615 620 Val Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile 625 630 635 640 Tyr Val Tyr Leu Ala Gly Cys Ser Ala Gln Val Val Asn Asp Leu Thr 645 650 655 Arg Asn Arg Phe Phe Glu Asn Pro Ala Leu Trp Glu Leu Leu Phe His 660 665 670 Ser Ile His Asp Ala Val Leu Gly Ser Gln Leu Arg Glu Ala Leu Ala 675 680 685 Glu Gln Glu Ala Ser Ala Pro Pro Ser Gln Glu Asp Leu Glu Pro Asn 690 695 700 Ala Thr Pro Ala Thr Pro Glu Ala 705 710 ＜210＞ 9 ＜211＞ 744 ＜212＞ PRT ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 9 Met Asp His Ala Glu Glu Asn Glu Ile Pro Ala Glu Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Val Lys Asp Lys Val Thr Glu Ser Ile Gly Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Val Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Phe Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Ser Leu His Glu Ser Tyr Ser Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Ile Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Leu Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Ala Val Leu Ser Ala Ile Val Ile Val Asn Leu 435 440 445 Lys Gly Met Phe Met Gln Phe Ser Asp Leu Pro Phe Phe Trp Arg Thr 450 455 460 Ser Lys Ile Glu Leu Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 465 470 475 480 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 485 490 495 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 500 505 510 Gln Leu Pro Asp Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 515 520 525 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 530 535 540 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Ser Ala Leu Lys Arg Lys Thr 545 550 555 560 Gly Val Asn Pro Ala Leu Ile Met Gly Ala Arg Arg Lys Ala Met Arg 565 570 575 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Val Ala Asn Ala Thr Val 580 585 590 Val Lys Val Asp Ala Glu Val Asp Gly Glu Asn Ala Thr Lys Pro Glu 595 600 605 Glu Glu Asp Asp Glu Val Lys Phe Pro Pro Ile Val Ile Lys Thr Thr 610 615 620 Phe Pro Glu Glu Leu Gln Arg Phe Leu Pro Gln Gly Glu Asn Val His 625 630 635 640 Thr Val Ile Leu Asp Phe Thr Gln Val Asn Phe Val Asp Ser Val Gly 645 650 655 Val Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile 660 665 670 Tyr Val Tyr Leu Ala Gly Cys Ser Pro Gln Val Val Asn Asp Leu Thr 675 680 685 Arg Asn Asn Phe Phe Glu Asn Pro Ala Leu Lys Glu Leu Leu Phe His 690 695 700 Ser Ile His Asp Ala Val Leu Gly Ser Gln Val Arg Glu Ala Met Ala 705 710 715 720 Glu Gln Glu Ala Thr Ala Ser Leu Pro Gln Glu Asp Met Glu Pro Asn 725 730 735 Ala Thr Pro Thr Thr Pro Glu Ala 740 ＜210＞ 10 ＜211＞ 744 ＜212＞ PRT ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 10 Met Asp His Ala Glu Glu Thr Glu Ile Pro Ala Glu Thr Gln Arg Tyr 1 5 10 15 Tyr Val Glu Arg Pro Ile Phe Ser His Pro Val Leu Gln Glu Arg Leu 20 25 30 His Val Lys Asp Lys Val Thr Glu Ser Ile Gly Asp Lys Leu Lys Gln 35 40 45 Ala Phe Thr Cys Thr Pro Lys Lys Ile Arg Asn Ile Ile Tyr Met Phe 50 55 60 Leu Pro Ile Thr Lys Trp Leu Pro Ala Tyr Lys Phe Lys Glu Tyr Val 65 70 75 80 Leu Gly Asp Leu Val Ser Gly Ile Ser Thr Gly Val Leu Gln Leu Pro 85 90 95 Gln Gly Leu Ala Phe Ala Met Leu Ala Ala Val Pro Pro Val Phe Gly 100 105 110 Leu Tyr Ser Ser Phe Tyr Pro Val Ile Met Tyr Cys Phe Phe Gly Thr 115 120 125 Ser Arg His Ile Ser Ile Gly Pro Phe Ala Val Ile Ser Leu Met Ile 130 135 140 Gly Gly Val Ala Val Arg Leu Val Pro Asp Asp Ile Val Ile Pro Gly 145 150 155 160 Gly Val Asn Ala Thr Asn Gly Thr Glu Ala Arg Asp Ala Leu Arg Val 165 170 175 Lys Val Ala Met Ser Val Thr Leu Leu Ser Gly Ile Ile Gln Phe Cys 180 185 190 Leu Gly Val Cys Arg Phe Gly Phe Val Ala Ile Tyr Leu Thr Glu Pro 195 200 205 Leu Val Arg Gly Phe Thr Thr Ala Ala Ala Val His Val Phe Thr Ser 210 215 220 Met Leu Lys Tyr Leu Phe Gly Val Lys Thr Lys Arg Tyr Ser Gly Ile 225 230 235 240 Phe Ser Val Val Tyr Ser Thr Val Ala Val Leu Gln Asn Val Lys Asn 245 250 255 Leu Asn Val Cys Ser Leu Gly Val Gly Leu Met Val Phe Gly Leu Leu 260 265 270 Leu Gly Gly Lys Glu Phe Asn Glu Arg Phe Lys Glu Lys Leu Pro Ala 275 280 285 Pro Ile Pro Leu Glu Phe Phe Ala Val Val Met Gly Thr Gly Ile Ser 290 295 300 Ala Gly Phe Ser Leu His Glu Ser Tyr Ser Val Asp Val Val Gly Thr 305 310 315 320 Leu Pro Leu Gly Leu Leu Pro Pro Ala Asn Pro Asp Thr Ser Leu Phe 325 330 335 His Leu Val Tyr Val Asp Ala Ile Ala Ile Ala Ile Val Gly Phe Ser 340 345 350 Val Thr Ile Ser Met Ala Lys Thr Leu Ala Asn Lys His Gly Tyr Gln 355 360 365 Val Asp Gly Asn Gln Glu Leu Ile Ala Leu Gly Ile Cys Asn Ser Ile 370 375 380 Gly Ser Leu Phe Gln Thr Phe Ser Ile Ser Cys Ser Leu Ser Arg Ser 385 390 395 400 Leu Val Gln Glu Gly Thr Gly Gly Lys Thr Gln Leu Ala Gly Cys Leu 405 410 415 Ala Ser Leu Met Ile Leu Leu Val Ile Leu Ala Thr Gly Phe Leu Phe 420 425 430 Glu Ser Leu Pro Gln Ala Val Leu Ser Ala Ile Val Ile Val Asn Leu 435 440 445 Lys Gly Met Phe Met Gln Phe Ser Asp Leu Pro Phe Phe Trp Arg Thr 450 455 460 Ser Lys Ile Glu Leu Thr Ile Trp Leu Thr Thr Phe Val Ser Ser Leu 465 470 475 480 Phe Leu Gly Leu Asp Tyr Gly Leu Ile Thr Ala Val Ile Ile Ala Leu 485 490 495 Leu Thr Val Ile Tyr Arg Thr Gln Ser Pro Ser Tyr Lys Val Leu Gly 500 505 510 Gln Leu Pro Asp Thr Asp Val Tyr Ile Asp Ile Asp Ala Tyr Glu Glu 515 520 525 Val Lys Glu Ile Pro Gly Ile Lys Ile Phe Gln Ile Asn Ala Pro Ile 530 535 540 Tyr Tyr Ala Asn Ser Asp Leu Tyr Ser Ser Ala Leu Lys Arg Lys Thr 545 550 555 560 Gly Val Asn Pro Ala Leu Ile Met Gly Ala Arg Arg Lys Ala Met Arg 565 570 575 Lys Tyr Ala Lys Glu Val Gly Asn Ala Asn Val Ala Asn Ala Thr Val 580 585 590 Val Lys Val Asp Ala Glu Val Asp Gly Glu Asn Ala Thr Lys Pro Glu 595 600 605 Glu Glu Asp Asp Glu Val Lys Phe Pro Pro Ile Val Ile Lys Thr Thr 610 615 620 Phe Pro Glu Glu Leu Gln Arg Phe Leu Pro Gln Gly Glu Asn Val His 625 630 635 640 Thr Val Ile Leu Asp Phe Thr Gln Val Asn Phe Val Asp Ser Val Gly 645 650 655 Val Lys Thr Leu Ala Gly Ile Val Lys Glu Tyr Gly Asp Val Gly Ile 660 665 670 Tyr Val Tyr Leu Ala Gly Cys Ser Pro Gln Val Val Asn Asp Leu Thr 675 680 685 Arg Asn Asn Phe Phe Glu Asn Pro Ala Leu Lys Glu Leu Leu Phe His 690 695 700 Ser Ile His Asp Ala Val Leu Gly Ser Gln Val Arg Glu Ala Met Ala 705 710 715 720 Glu Gln Glu Ala Thr Ala Ser Leu Pro Gln Glu Asp Met Glu Pro Asn 725 730 735 Ala Thr Pro Thr Thr Pro Glu Ala 740 ＜210＞ 11 ＜211＞ 3848 ＜212＞ DNA ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 11 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 60 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 120 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 180 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 240 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg 300 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 360 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 420 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 480 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt 540 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 600 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 660 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 720 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa 780 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca 840 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca 900 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 960 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctct 1020 gcagaccatg gatcatgctg aagaaaatga aatccctgca gagacccaga ggtactacgt 1080 ggaaaggccc atcttcagtc atcctgtcct ccaagagagg ctgcacgtca aggacaaagt 1140 cacagagtcc attggagata agctgaagca ggcattcacg tgtactccta aaaaaataag 1200 aaacatcatt tacatgttcc tgcctatcac taagtggctg ccagcatata aattcaagga 1260 gtatgtgtta ggtgacttgg tctcgggcat aagcactggg gtactccagc ttccccaagg 1320 cttagccttc gccatgctgg cagccgtgcc tccggtgttt ggcctgtact catcgtttta 1380 ccccgttatc atgtactgtt tctttggaac ctcaagacac atatctatag gtccttttgc 1440 tgttattagc ttgatgattg gaggtgtggc cgtccggtta gtaccagatg atattgtcat 1500 cccaggagga gtaaatgcaa ccaacgggac agaagccaga gatgcactaa gagtgaaagt 1560 cgccatgtct gttaccttac tttcaggaat cattcagttt tgcctaggtg tctgtaggtt 1620 tggatttgtg gccatatacc tcacggagcc attggtgcga ggctttacca ctgcggctgc 1680 tgtccacgtg ttcacgtcca tgttaaaata cctgtttggg gtcaaaacaa agcggtacag 1740 tggaatcttt tcagtggtgt atagtacagt tgctgtgttg cagaatgtta aaaacctcaa 1800 cgtgtgttcc ctaggcgtcg gcctgatggt ttttggtttg ctgttgggtg gcaaggaatt 1860 taatgagaga tttaaagaga aattgccagc acccattcct ctagagttct ttgctgtggt 1920 gatggggact ggcatttctg caggatttaa cctacatgag tcctacagtg tggatgtcgt 1980 tggaacactt cctctggggc tacttcctcc ggccaaccca gacaccagcc tgttccacct 2040 ggtgtatgtg gacgccattg ccatcgccat cgttggattt tcagtgacga tctccatggc 2100 caaaaccttg gcaaataagc atggctacca ggttgatggc aatcaggagc tcattgcctt 2160 ggggatatgc aactccattg gatctctctt ccaaaccttc tcgatttcct gctccttgtc 2220 tcgaagcctt gttcaggaag gaactggagg gaaaacacag cttgcaggtt gtttggcctc 2280 gttgatgatt ctgttggtca tattagccac cggattcctc tttgagtcgt taccccaggc 2340 tgtcctttcc gccattgtga tcgtcaacct gaaaggaatg ttcatgcagt tctcagacct 2400 gccttttttc tggagaacca gcaaaataga gctgaccatc tggctgacca cctttgtgtc 2460 ctccctgttc ctcggcttgg actacggact gattaccgcc gtgatcattg ctctgctcac 2520 agtgatttat agaacacaga gtccaagcta caaagtcctg gggcagctcc ctgacacgga 2580 tgtgtacatt gacatagatg catatgagga ggtgaaagaa attcctggaa taaaaatatt 2640 ccaaataaat gccccaattt actatgcaaa tagcgacttg tatagcagcg ctttaaaaag 2700 aaagactgga gtaaacccag cactcattat gggagcgaga agaaaggcca tgaggaagta 2760 cgccaaggaa gttggaaatg ccaacgtggc caatgctact gttgtcaaag tggatgcaga 2820 agtagacgga gaaaatgcta caaaacctga agaagaggat gatgaagtca aatttccccc 2880 aatagtcatc aaaacaacat ttcctgaaga gctgcagaga tttttgcccc agggggaaaa 2940 tgtccacact gtcattctag actttacgca ggtcaatttt gtggattctg ttggagtgaa 3000 aactctggcc gggattgtga aagaatatgg agatgttgga atttatgtat atttagcagg 3060 atgcagccca caagttgtga atgacctcac ccgcaacaac ttttttgaaa atcctgcctt 3120 gaaagagctt ctgttccaca gtatccacga tgcagtcctg ggcagccaag ttcgggaggc 3180 aatggctgaa caagaagcca cagcgtcact tccccaggag gatatggagc ccaatgccac 3240 acccaccacc cccgaggcat aactcgaggg ggggcccggt accgcgggcc cgggatccac 3300 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 3360 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 3420 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 3480 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 3540 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa 3600 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat 3660 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg 3720 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac 3780 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta 3840 aagggagc 3848 ＜210＞ 12 ＜211＞ 3848 ＜212＞ DNA ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 12 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 60 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 120 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 180 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 240 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg 300 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 360 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 420 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 480 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt 540 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 600 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 660 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 720 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa 780 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca 840 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca 900 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 960 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctct 1020 gcagaccatg gatcatgctg aagaaaccga aatccctgca gagacccaga ggtactacgt 1080 ggaaaggccc atcttcagtc atcctgtcct ccaagagagg ctgcacgtca aggacaaagt 1140 cacagagtcc attggagata agctgaagca ggcattcacg tgtactccta aaaaaataag 1200 aaacatcatt tacatgttcc tgcctatcac taagtggctg ccagcatata aattcaagga 1260 gtatgtgtta ggtgacttgg tctcgggcat aagcactggg gtactccagc ttccccaagg 1320 cttagccttc gccatgctgg cagccgtgcc tccggtgttt ggcctgtact catcgtttta 1380 ccccgttatc atgtactgtt tctttggaac ctcaagacac atatctatag gtccttttgc 1440 tgttattagc ttgatgattg gaggtgtggc cgtccggtta gtaccagatg atattgtcat 1500 cccaggagga gtaaatgcaa ccaacgggac agaagccaga gatgcactaa gagtgaaagt 1560 cgccatgtct gttaccttac tttcaggaat cattcagttt tgcctaggtg tctgtaggtt 1620 tggatttgtg gccatatacc tcacggagcc attggtgcga ggctttacca ctgcggctgc 1680 tgtccacgtg ttcacgtcca tgttaaaata cctgtttggg gtcaaaacaa agcggtacag 1740 tggaatcttt tcagtggtgt atagtacagt tgctgtgttg cagaatgtta aaaacctcaa 1800 cgtgtgttcc ctaggcgtcg gcctgatggt ttttggtttg ctgttgggtg gcaaggaatt 1860 taatgagaga tttaaagaga aattgccagc acccattcct ctagagttct ttgctgtggt 1920 gatggggact ggcatttctg caggatttaa cctacatgag tcctacagtg tggatgtcgt 1980 tggaacactt cctctggggc tacttcctcc ggccaaccca gacaccagcc tgttccacct 2040 ggtgtatgtg gacgccattg ccatcgccat cgttggattt tcagtgacga tctccatggc 2100 caaaaccttg gcaaataagc atggctacca ggttgatggc aatcaggagc tcattgcctt 2160 ggggatatgc aactccattg gatctctctt ccaaaccttc tcgatttcct gctccttgtc 2220 tcgaagcctt gttcaggaag gaactggagg gaaaacacag cttgcaggtt gtttggcctc 2280 gttgatgatt ctgttggtca tattagccac cggattcctc tttgagtcgt taccccaggc 2340 tgtcctttcc gccattgtga tcgtcaacct gaaaggaatg ttcatgcagt tctcagacct 2400 gccttttttc tggagaacca gcaaaataga gctgaccatc tggctgacca cctttgtgtc 2460 ctccctgttc ctcggcttgg actacggact gattaccgcc gtgatcattg ctctgctcac 2520 agtgatttat agaacacaga gtccaagcta caaagtcctg gggcagctcc ctgacacgga 2580 tgtgtacatt gacatagatg catatgagga ggtgaaagaa attcctggaa taaaaatatt 2640 ccaaataaat gccccaattt actatgcaaa tagcgacttg tatagcagcg ctttaaaaag 2700 aaagactgga gtaaacccag cactcattat gggagcgaga agaaaggcca tgaggaagta 2760 cgccaaggaa gttggaaatg ccaacgtggc caatgctact gttgtcaaag tggatgcaga 2820 agtagacgga gaaaatgcta caaaacctga agaagaggat gatgaagtca aatttccccc 2880 aatagtcatc aaaacaacat ttcctgaaga gctgcagaga tttttgcccc agggggaaaa 2940 tgtccacact gtcattctag actttacgca ggtcaatttt gtggattctg ttggagtgaa 3000 aactctggcc gggattgtga aagaatatgg agatgttgga atttatgtat atttagcagg 3060 atgcagccca caagttgtga atgacctcac ccgcaacaac ttttttgaaa atcctgcctt 3120 gaaagagctt ctgttccaca gtatccacga tgcagtcctg ggcagccaag ttcgggaggc 3180 aatggctgaa caagaagcca cagcgtcact tccccaggag gatatggagc ccaatgccac 3240 acccaccacc cccgaggcat aactcgaggg ggggcccggt accgcgggcc cgggatccac 3300 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 3360 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 3420 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 3480 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 3540 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa 3600 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat 3660 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg 3720 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac 3780 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta 3840 aagggagc 3848 ＜210＞ 13 ＜211＞ 3379 ＜212＞ DNA ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 13 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 60 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 120 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 180 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 240 ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg tgagcaaggg 300 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 360 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 420 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 480 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt 540 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 600 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 660 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 720 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa 780 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca 840 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca 900 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 960 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac tcagatctct 1020 gcagaccatg gatcatgctg aagaaaatga aatccctgca gagacccaga ggtactacgt 1080 ggaaaggccc atcttcagtc atcctgtcct ccaagagagg ctgcacgtca aggacaaagt 1140 cacagagtcc attggagata agctgaagca ggcattcacg tgtactccta aaaaaataag 1200 aaacatcatt tacatgttcc tgcctatcac taagtggctg ccagcatata aattcaagga 1260 gtatgtgtta ggtgacttgg tctcgggcat aagcactggg gtactccagc ttccccaagg 1320 cttagccttc gccatgctgg cagccgtgcc tccggtgttt ggcctgtact catcgtttta 1380 ccccgttatc atgtactgtt tctttggaac ctcaagacac atatctatag gtccttttgc 1440 tgttattagc ttgatgattg gaggtgtggc cgtccggtta gtaccagatg atattgtcat 1500 cccaggagga gtaaatgcaa ccaacgggac agaagccaga gatgcactaa gagtgaaagt 1560 cgccatgtct gttaccttac tttcaggaat cattcagttt tgcctaggtg tctgtaggtt 1620 tggatttgtg gccatatacc tcacggagcc attggtgcga ggctttacca ctgcggctgc 1680 tgtccacgtg ttcacgtcca tgttaaaata cctgtttggg gtcaaaacaa agcggtacag 1740 tggaatcttt tcagtggtgt atagtacagt tgctgtgttg cagaatgtta aaaacctcaa 1800 cgtgtgttcc ctaggcgtcg gcctgatggt ttttggtttg ctgttgggtg gcaaggaatt 1860 taatgagaga tttaaagaga aattgccagc acccattcct ctagagttct ttgctgtggt 1920 gatggggact ggcatttctg caggattttc cctacatgag tcctacagtg tggatgtcgt 1980 tggaacactt cctctggggc tacttcctcc ggccaaccca gacaccagcc tgttccacct 2040 ggtgtatgtg gacgccattg ccatcgccat cgttggattt tcagtgacga tctccatggc 2100 caaaaccttg gcaaataagc atggctacca ggttgatggc aatcaggagc tcattgcctt 2160 ggggatatgc aactccattg gatctctctt ccaaaccttc tcgatttcct gctccttgtc 2220 tcgaagcctt gttcaggaag gaactggagg gaaaacacag cttgcaggtt gtttggcctc 2280 gttgatgatt ctgttggtca tattagccac cggattcctc tttgagtcgt taccccaggc 2340 tgtcctttcc gccattgtga tcgtcaacct gaaaggaatg ttcatgcagt tctcagacct 2400 gccttttttc tggagaacca gcaaaataga gctgaccatc tggctgacca cctttgtgtc 2460 ctccctgttc ctcggcttgg actacggact gattaccgcc gtgatcattg ctctgctcac 2520 agtgatttat agaacacaga gtccaagcta caaagtcctg gggcagctcc ctgacacgga 2580 tgtgtacatt gacatagatg catatgagga ggtgaaagaa attcctggaa taaaaatatt 2640 ccaaataaat gccccaattt actatgcaaa tagcgacttg tatagcagcg ctttaaaaag 2700 aaagactgga gtaaacccag cactcattat gggagcgaga agaaaggcca tgaggaagta 2760 cgccaaggaa gttggaaatg ccaacgtggc caatgctact gttgtcaaag tggatgcaga 2820 agtagacgga gaaaatgcta caaaacctga agaagaggat gatgaagtca aatttccccc 2880 aatagtcatc aaaacaacat ttcctgaaga gctgcagaga tttttgcccc agggggaaaa 2940 tgtccacact gtcattctag actttacgca ggtcaatttt gtggattctg ttggagtgaa 3000 aactctggcc gggattgtga aagaatatgg agatgttgga atttatgtat atttagcagg 3060 atgcagccca caagttgtga atgacctcac ccgcaacaac ttttttgaaa atcctgcctt 3120 gaaagagctt ctgttccaca gtatccacga tgcagtcctg ggcagccaag ttcgggaggc 3180 aatggctgaa caagaagcca cagcgtcact tccccaggag gatatggagc ccaatgccac 3240 acccaccacc cccgaggcat aactcgaggg ggggcccggt accgcgggcc cgggatccac 3300 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 3360 aaaaaacctc ccacacctc 3379 ＜210＞ 14 ＜211＞ 3705 ＜212＞ DNA ＜213＞ artificial sequence ＜220＞ ＜223＞ synthetic sequence ＜400＞ 14 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 60 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 120 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 180 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 240 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 300 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 360 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 420 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 480 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 540 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 600 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 660 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 720 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg 780 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc 840 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 900 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 960 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1020 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac 1080 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc 1140 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1200 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc 1260 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1320 ctgtacaagt ccggactcag atctctgcag accatggatc atgctgaaga aaccgaaatc 1380 cctgcagaga cccagaggta ctacgtggaa aggcccatct tcagtcatcc tgtcctccaa 1440 gagaggctgc acgtcaagga caaagtcaca gagtccattg gagataagct gaagcaggca 1500 ttcacgtgta ctcctaaaaa aataagaaac atcatttaca tgttcctgcc tatcactaag 1560 tggctgccag catataaatt caaggagtat gtgttaggtg acttggtctc gggcataagc 1620 actggggtac tccagcttcc ccaaggctta gccttcgcca tgctggcagc cgtgcctccg 1680 gtgtttggcc tgtactcatc gttttacccc gttatcatgt actgtttctt tggaacctca 1740 agacacatat ctataggtcc ttttgctgtt attagcttga tgattggagg tgtggccgtc 1800 cggttagtac cagatgatat tgtcatccca ggaggagtaa atgcaaccaa cgggacagaa 1860 gccagagatg cactaagagt gaaagtcgcc atgtctgtta ccttactttc aggaatcatt 1920 cagttttgcc taggtgtctg taggtttgga tttgtggcca tatacctcac ggagccattg 1980 gtgcgaggct ttaccactgc ggctgctgtc cacgtgttca cgtccatgtt aaaatacctg 2040 tttggggtca aaacaaagcg gtacagtgga atcttttcag tggtgtatag tacagttgct 2100 gtgttgcaga atgttaaaaa cctcaacgtg tgttccctag gcgtcggcct gatggttttt 2160 ggtttgctgt tgggtggcaa ggaatttaat gagagattta aagagaaatt gccagcaccc 2220 attcctctag agttctttgc tgtggtgatg gggactggca tttctgcagg attttcccta 2280 catgagtcct acagtgtgga tgtcgttgga acacttcctc tggggctact tcctccggcc 2340 aacccagaca ccagcctgtt ccacctggtg tatgtggacg ccattgccat cgccatcgtt 2400 ggattttcag tgacgatctc catggccaaa accttggcaa ataagcatgg ctaccaggtt 2460 gatggcaatc aggagctcat tgccttgggg atatgcaact ccattggatc tctcttccaa 2520 accttctcga tttcctgctc cttgtctcga agccttgttc aggaaggaac tggagggaaa 2580 acacagcttg caggttgttt ggcctcgttg atgattctgt tggtcatatt agccaccgga 2640 ttcctctttg agtcgttacc ccaggctgtc ctttccgcca ttgtgatcgt caacctgaaa 2700 ggaatgttca tgcagttctc agacctgcct tttttctgga gaaccagcaa aatagagctg 2760 accatctggc tgaccacctt tgtgtcctcc ctgttcctcg gcttggacta cggactgatt 2820 accgccgtga tcattgctct gctcacagtg atttatagaa cacagagtcc aagctacaaa 2880 gtcctggggc agctccctga cacggatgtg tacattgaca tagatgcata tgaggaggtg 2940 aaagaaattc ctggaataaa aatattccaa ataaatgccc caatttacta tgcaaatagc 3000 gacttgtata gcagcgcttt aaaaagaaag actggagtaa acccagcact cattatggga 3060 gcgagaagaa aggccatgag gaagtacgcc aaggaagttg gaaatgccaa cgtggccaat 3120 gctactgttg tcaaagtgga tgcagaagta gacggagaaa atgctacaaa acctgaagaa 3180 gaggatgatg aagtcaaatt tcccccaata gtcatcaaaa caacatttcc tgaagagctg 3240 cagagatttt tgccccaggg ggaaaatgtc cacactgtca ttctagactt tacgcaggtc 3300 aattttgtgg attctgttgg agtgaaaact ctggccggga ttgtgaaaga atatggagat 3360 gttggaattt atgtatattt agcaggatgc agcccacaag ttgtgaatga cctcacccgc 3420 aacaactttt ttgaaaatcc tgccttgaaa gagcttctgt tccacagtat ccacgatgca 3480 gtcctgggca gccaagttcg ggaggcaatg gctgaacaag aagccacagc gtcacttccc 3540 caggaggata tggagcccaa tgccacaccc accacccccg aggcataact cgaggggggg 3600 cccggtaccg cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc 3660 acatttgtag aggttttact tgctttaaaa aacctcccac acctc 3705
Figure 12_A0101_SEQ_0001
Figure 12_A0101_SEQ_0002
Figure 12_A0101_SEQ_0003
Figure 12_A0101_SEQ_0004
Figure 12_A0101_SEQ_0005
Figure 12_A0101_SEQ_0006
Figure 12_A0101_SEQ_0007
Figure 12_A0101_SEQ_0008
Figure 12_A0101_SEQ_0009
Figure 12_A0101_SEQ_0010
Figure 12_A0101_SEQ_0011
Figure 12_A0101_SEQ_0012
Figure 12_A0101_SEQ_0013
Figure 12_A0101_SEQ_0014
Figure 12_A0101_SEQ_0015
Figure 12_A0101_SEQ_0016
Figure 12_A0101_SEQ_0017
Figure 12_A0101_SEQ_0018
Figure 12_A0101_SEQ_0019
Figure 12_A0101_SEQ_0020
Figure 12_A0101_SEQ_0021
Figure 12_A0101_SEQ_0022
Figure 12_A0101_SEQ_0023
Figure 12_A0101_SEQ_0024
Figure 12_A0101_SEQ_0025
Figure 12_A0101_SEQ_0026
Figure 12_A0101_SEQ_0027
Figure 12_A0101_SEQ_0028
Figure 12_A0101_SEQ_0029
Figure 12_A0101_SEQ_0030
Figure 12_A0101_SEQ_0031
Figure 12_A0101_SEQ_0032
Figure 12_A0101_SEQ_0033
Figure 12_A0101_SEQ_0034
Figure 12_A0101_SEQ_0035
Figure 12_A0101_SEQ_0036
Figure 12_A0101_SEQ_0037
Figure 12_A0101_SEQ_0038
Figure 12_A0101_SEQ_0039
Figure 12_A0101_SEQ_0040
Figure 12_A0101_SEQ_0041
Figure 12_A0101_SEQ_0042
Figure 12_A0101_SEQ_0043
Figure 12_A0101_SEQ_0044
Figure 12_A0101_SEQ_0045
Figure 12_A0101_SEQ_0046
Figure 12_A0101_SEQ_0047
Figure 12_A0101_SEQ_0048
Figure 12_A0101_SEQ_0049
Figure 12_A0101_SEQ_0050
Figure 12_A0101_SEQ_0051
Figure 12_A0101_SEQ_0052
Figure 12_A0101_SEQ_0053
Figure 12_A0101_SEQ_0054
Figure 12_A0101_SEQ_0055
Figure 12_A0101_SEQ_0056

Claims (9)

  1. 一種超音波感應蛋白質，其係具有無法感受超音波的哺乳動物耳蝸外毛細胞的馬達蛋白胺基酸序列第308個位置的天門冬醯胺被絲氨酸取代的變異(N308S)、以及選擇性地具有第7個位置的天門冬醯胺被蘇胺酸取代的變異(N7T)。
  2. 如申請專利範圍第1項所述之超音波感應蛋白質，其中，該無法感受超音波的哺乳動物的耳蝸外毛細胞的馬達蛋白係人類、小鼠、狐蝠、小鬚鯨、長舌果蝠、或犬蝠的耳蝸外毛細胞的馬達蛋白。
  3. 如申請專利範圍第2項所述之超音波感應蛋白質，其中，人類的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 1；小鼠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 2；狐蝠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 3；小鬚鯨的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 4；長舌果蝠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 5；以及，犬蝠的耳蝸外毛細胞的馬達蛋白的胺基酸序列為SEQ ID NO: 6。
  4. 如申請專利範圍第3項所述之超音波感應蛋白質，其中，人類的耳蝸外毛細胞的N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO: 7；人類的耳蝸外毛細胞的N7T、N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO: 8；小鼠的耳蝸外毛細胞的N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO: 9; 小鼠的耳蝸外毛細胞的N7T、N308S變異型馬達蛋白的胺基酸序列為SEQ ID NO:10。
  5. 一種刺激細胞的方法，包括利用一超音波照射表現一超音波感應蛋白質的一細胞，其中，該超音波感應蛋白質具有無法感受超音波的哺乳動物的耳蝸外毛細胞的馬達蛋白胺基酸序列第308個位置的天門冬醯胺被絲氨酸取代的變異(N308S)、以及選擇性地具有第7個位置的天門冬醯胺被蘇胺酸取代的變異(N7T)。
  6. 如申請專利範圍第5項所述之方法，其中，該超音波係一對焦式超音波。
  7. 如申請專利範圍第6項所述之方法，其中，該超音波發射頻率為0.5 MHz、聲壓為0.5 MPa。
  8. 如申請專利範圍第5項所述之方法，其中，該細胞經該超音波照射後，被誘導鈣離子流入該細胞內。
  9. 如申請專利範圍第5項所述之方法，其中，該細胞為神經細胞、免疫細胞、胰島細胞、上皮細胞、血液細胞、肌肉細胞、或幹細胞。
TW107120094A 2018-06-12 2018-06-12 超音波感應蛋白質以及利用超音波刺激細胞的方法 TWI720319B (zh)

Priority Applications (2)

Application Number Priority Date Filing Date Title
TW107120094A TWI720319B (zh) 2018-06-12 2018-06-12 超音波感應蛋白質以及利用超音波刺激細胞的方法
US16/290,214 US20190374763A1 (en) 2018-06-12 2019-03-01 Ultrasound-sensing proteins and method for stimulating cells by ultrasound

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
TW107120094A TWI720319B (zh) 2018-06-12 2018-06-12 超音波感應蛋白質以及利用超音波刺激細胞的方法

Publications (2)

Publication Number Publication Date
TW202000918A true TW202000918A (zh) 2020-01-01
TWI720319B TWI720319B (zh) 2021-03-01

Family

ID=68764112

Family Applications (1)

Application Number Title Priority Date Filing Date
TW107120094A TWI720319B (zh) 2018-06-12 2018-06-12 超音波感應蛋白質以及利用超音波刺激細胞的方法

Country Status (2)

Country Link
US (1) US20190374763A1 (zh)
TW (1) TWI720319B (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TWI810633B (zh) * 2020-08-24 2023-08-01 國立清華大學 超音波搭配微氣泡輔助之氣球導管系統及其血管擴張方法

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6602992B1 (en) * 2000-02-18 2003-08-05 Northwestern University Mammalian prestin polynucleotides
US10806951B2 (en) * 2014-09-24 2020-10-20 Salk Institute For Biological Studies Sonogenic stimulation of cells

Also Published As

Publication number Publication date
US20190374763A1 (en) 2019-12-12
TWI720319B (zh) 2021-03-01

Similar Documents

Publication Publication Date Title
CN111228475A (zh) 用于预防新型冠状病毒的生物制品
CN107759701B (zh) 嵌合抗原受体、其修饰的NK细胞、编码DNA、mRNA、表达载体、制备方法和应用
CN110295149B (zh) 一种突变株3型鸭甲肝病毒ch-p60-117c株及构建方法
KR20190042473A (ko) 중증 혈소판 감소 증후군 바이러스 예방 및 치료용 유전자 백신
TWI720319B (zh) 超音波感應蛋白質以及利用超音波刺激細胞的方法
CN110218732B (zh) 一种非洲猪瘟病毒串联基因、共表达载体、构建方法及应用
CN112391412A (zh) 一种调控脂质代谢的复制型溶瘤腺病毒及其应用
CN101121939B (zh) siRNA筛选系统的通用性绿色荧光蛋白融合靶基因表达载体
CN109055375A (zh) 一种crispr辅助反式增强子激活基因表达的方法及其应用
CN109456991A (zh) 原儿茶酸调控的开关系统及其调控方法和应用
CN110295180B (zh) 一种3型鸭甲肝病毒突变基因isa-a117c-c4334a及构建方法
CN110684781B (zh) 一种3型鸭甲肝病毒突变基因isa-a117c-t1142a及构建方法
CA3226725A1 (en) Composition and uses thereof
CN109456992B (zh) 原儿茶酸调控的多功能基因表达平台及其应用
KR102599298B1 (ko) 전기천공을 사용하여 지방 조직의 생체내 형질감염을 최소로 침습성으로 하기 위한 방법 및 장치
CN110484546B (zh) 一种3型鸭甲肝病毒突变基因isa-c4334a及构建方法
CN114395017A (zh) SARS-CoV-2病毒样颗粒的制备方法及其应用
CN100386437C (zh) 高表达水平的转基因动物乳腺特异性载体的构建方法
CN110283835B (zh) 一种3型鸭甲肝病毒突变基因isa-t1142a-c4334a及构建方法
KR102067487B1 (ko) 베타-디펜신 이합체를 포함하는 신규 세포투과성 펩타이드 및 이의 용도
CN111518836A (zh) 一种用于体内基因表达蛋白系统的转染剂及其体外转染的方法
CN114010666B (zh) 溶瘤病毒、parp抑制剂及pd-1抗体在制备抗肿瘤药物中的应用
CN110283834B (zh) 一种3型鸭甲肝病毒突变基因isa-a117c及构建方法
CN110283836B (zh) 一种3型鸭甲肝病毒突变基因isa-t1142a及构建方法
CN113215116B (zh) 基于朝阳病毒载体的嵌合病毒、疫苗及其应用