具体实施方式
在下文中,将详细描述本发明。
但是,以下实施例用于说明本发明,并且本发明不限于以下实施例。
实施例1:层粘连蛋白信号在免疫细胞迁移和侵袭中的作用的验证
在构成血管的细胞外基质(ECM)中,已识别出是什么促进单核细胞/巨噬细胞的迁移和侵袭。使用胶原蛋白(胶原蛋白,Col),纤维连接蛋白(FN)和层粘连蛋白(LN)作为细胞外基质进行Transwell迁移实验。具体的实验方法如下。用明胶(0.5mg/ml)包被Transwell(Corning,#3421-5mm)后,将RAW 264.7细胞(1×105个细胞/孔)接种到上室(top chamber)中。将纤维连接蛋白或胶原蛋白置于下室(bottom chamber)中。将分别含有10μg/ml层粘连蛋白(层粘连蛋白混合物,Biolamina)、纤维连接蛋白或胶原蛋白的无血清DMEM(500μl)置于下室中。24小时后,将细胞用70%甲醇处理30分钟并固定,然后用50%苏木精染色30分钟。用棉签从膜上除去未迁移的细胞后,取出膜并将其装在载玻片上。在高倍率显微镜下观察并定量膜下侧的迁移细胞。
如图1和2所示,证实了各种细胞外基质中的层粘连蛋白极大地促进了单核细胞/巨噬细胞的迁移。因此,证实了单核细胞/巨噬细胞的迁移对细胞外基质(ECM)中的层粘连蛋白(LN)信号反应最强烈。
实施例2:层粘连蛋白亚型对免疫细胞迁移和侵袭的影响
评估了层粘连蛋白亚型对免疫细胞迁移和侵袭的影响。以与实施例1中相同的方式,使用1μg/ml的各层粘连蛋白亚型蛋白LN111、LN211、LN221、LN411、LN421、LN511和LN521(购自Biolamina),进行Transwell细胞迁移实验。根据由每种层粘连蛋白亚型组成的链,层粘连蛋白亚型的具体序列可参考SEQ ID NO:120的α4链、SEQ ID NO:126的α2链、SEQ IDNO:127的α5链、SEQ ID NO:122的β2链、SEQ ID NO:128的β1链、SEQ ID NO:124的γ1链。
如图3和图4所示,证实了单核细胞/巨噬细胞特别响应层粘连蛋白亚型中的α4β2γ1亚型(LN421)而运动。因此,证实了单核细胞/巨噬细胞的迁移对层粘连蛋白亚型中的LN421具有特异性。
实施例3:层粘连蛋白处理免疫细胞后细胞质KRS向细胞膜的运动_免疫细胞迁移相关疾病的新病理学的确定
将RAW 264.7细胞(2×106细胞)接种到100饼盘(pie plate)中并孵育18小时后,用1μg/ml的LN421处理无血清DMEM培养基,并在0h、12h和24h收获细胞。使用亚细胞蛋白质组提取试剂盒(Calbiotech,目录号539790)将RAW 264.7细胞蛋白分为胞质溶胶和膜成分。对获得的蛋白质进行电泳,转移至PVDF膜(Millipore),并用3%脱脂乳封闭。然后通过蛋白质印迹法检测KRS。具体地,加入KRS多克隆抗体(兔,Neomics,Co.Ltd.NMS-01-0005)以结合1小时。除去未结合的抗体,然后加入抗兔二抗(ThermoFisher Scientific,#31460)。与二抗反应后,使用ECL试剂作为底物在暗室中显影该膜。将检测到的条带与标准分子标记进行比较,以识别对应于KRS大小的条带。针对Na+/K+ATP酶(Abcam,ab76020)和微管蛋白(Santa cruz SC-5286)的抗体分别用于鉴定质膜和胞质溶胶标记物。
如图5所示,证实了当用LN421处理单核细胞/巨噬细胞时,在细胞膜区域中检测到的KRS量增加了,而在胞质溶胶区域中检测到的KRS量部分地减少了。这些结果表明,在单核细胞/巨噬细胞的细胞中表达并通常存在于细胞质区域中的KRS经过LN421处理后迁移到细胞膜区域。在免疫细胞膜区域中KRS特别增加,这被认为是在与免疫细胞迁移和侵袭关联的疾病中的重要的病理现象。
实施例4:通过降低细胞膜位置中的KRS水平对免疫细胞迁移/侵袭的控制以及免疫细胞迁移相关疾病的治疗效果的证实_抑制KRS迁移至细胞膜的化合物的用途
本发明人从上述实施例的结果证实,KRS的细胞内行为显著地影响单核细胞/巨噬细胞的迁移。特别地,随着KRS向细胞膜移动,KRS在免疫细胞膜区域特别增加的现象被认为是免疫细胞迁移和侵袭相关疾病的重要的病理学。因此,本发明人证实了抑制KRS的这种病理现象可用作免疫细胞迁移和侵袭有关疾病的治疗策略之一,这已经在韩国申请10-2017-0076718和10-2018-0069146中公开了。
在上述文件中,本发明人发现了抑制KRS向细胞膜移动的化合物,以抑制免疫细胞迁移相关疾病中免疫细胞膜的KRS的增加,以及证实和核查了它们对免疫细胞迁移相关疾病的治疗效果。
简而言之,为了抑制免疫细胞膜的KRS的增加,发现可使用BC-KI-00053(4-({(7-氟苯并[d]噻唑-2-基][2-(4-甲氧苯基)乙基]氨基}甲基)苯甲酸等化合物作为KRS迁移到细胞膜的抑制剂的代表性实例。在急性炎症反应(例如耳朵皮肤伤口模型)和缺血性免疫反应(例如肝脏缺血-再灌注损伤模型)等的炎症病况下,还证实了通过施用KRS迁移到细胞膜的抑制剂(特别是BC-KI-00053)可抑制免疫细胞迁移和侵袭,且出现了缓解疾病的效果。另外,在免疫细胞迁移相关疾病(例如肝纤维化、肺动脉高压(PAH)、高血压、蛋白尿、肾小球硬化症、肾纤维化和心肌纤维化以及Alport综合征)的体内模型中,证实了由免疫细胞迁移和侵袭引起的疾病状态,在此还证实了KRS迁移到细胞膜的抑制性化合物的治疗疾病的效果。
实施例5:用于降低细胞膜KRS水平的抗体的构建和免疫细胞迁移/侵袭控制作用的验证
5-1.特异性结合KRS N-末端的抗体_N3抗体的构建
由于实施例4的化合物通过抑制细胞膜部位的KRS水平的升高而显示出治疗和改善免疫细胞迁移/侵袭相关疾病的效果,本发明人试图基于相似原理生产表现优良的治疗效果的抗体。另一方面,本发明人发现了,当KRS从细胞质中移出并位于细胞膜中时,一些N-末端区域也暴露于细胞外膜(通常是KRS N-末端1~72的氨基酸区域)。因此,在抗KRS抗体中,能够与KRS-N末端结合的抗体被认为在体内对免疫细胞的迁移/侵袭的抑制具有更显著的优势,且作为代表的实施例,本发明人证实了通过如下所述的N3抗体的构建获得这些治疗优势。通过以下方法获得N3抗体。
具体地,当通过源自人KRS全长序列(SEQ ID NO:118)的层粘连蛋白信号移动至细胞膜时,选择仅与暴露于外膜的人KRS N-末端(SEQ ID NO:148)区域特异性结合的scFv,使用源自用HA标签标记的人B细胞的scFv噬菌体文库进行噬菌体展示淘选实验。在韩国专利号10-0961392中描述了在本实验中使用的scFv展示噬菌体文库(文库大小:约7.6×109,Hyunbo Shim教授生产的文库)。人KRS全长序列和N-末端位置的不同特定区域的KRS片段被用作噬菌体展示淘选实验的抗原蛋白。
噬菌体展示淘选实验具体如下进行。将1-10μg的抗原蛋白加入含有1ml 1×PBS溶液的免疫管中,并通过在37℃和200rpm下反应1小时将抗原包被在管的内表面上。排干抗原溶液,并用自来水洗涤1次以除去未包被的抗原。为了防止抗原蛋白和噬菌体之间的非特异性结合,在室温下使免疫管和scFv文库与含有3%脱脂乳的1×PBST(含有0.05%吐温20的PBS)反应1小时。从免疫管中去除脱脂牛奶后,加入scFv文库并在37℃和150rpm下反应1小时,以使scFv噬菌体与抗原结合。在室温下,通过加入1ml三乙胺(100mM)在10分钟内分离与每种抗原特异性结合的scFv噬菌体,并用Tris(1M,pH 7.4)中和。将过滤的噬菌体scFv加入到OD<1的孵育的ER2537大肠杆菌中,然后在37℃和120rpm孵育1小时30分钟时对其进行感染。将被噬菌体感染的大肠杆菌离心以去除一些培养上清液,然后再分散以覆盖到直径为15cm的含有氨苄青霉素(ampicillin)和葡萄糖(2%)的琼脂糖平板上。第二天,覆盖5ml SB培养基,以得到所有在平板上生长的细胞,然后将甘油(50%)添加至总体积的0.5倍,混合并各等分1ml,在-80℃下保存(scFv淘选储备液)。在20ml SB溶液中接种20μl制备的储备液并进行培养,然后使用辅助噬菌体(helper phage)制备scFv噬菌体文库(1ml)用于下一步的噬菌体淘选。重复上述步骤2-3次以分离表达抗原特异性scFv的噬菌体。
生物淘选后,进一步进行蛋白质印迹和免疫沉淀以选择与靶蛋白具有高结合能力的scFv克隆。
将选择的scFv转化为IgG(总抗体),具体方法如下。首先,通过PCR扩增在scFv克隆的基因组中编码scFv的多核苷酸。用于扩增scFv的VH区域的基因的引物的碱基序列如下:正向(AGA GAG TGT ACA CTC C CA GGC GGC CGA GGT GCA G,SEQ ID NO:129),和反向(CGCCGC TGG GCC CTT GGT GGA GGC)TGA GCT CAC GGT GAC CAG,SEQ ID NO:130)。用于扩增scFv的VL区域的基因的引物的核苷酸序列如下:正向(AAG CGG CCG CCA CCA TGG GAT GGAGCT GTA TCA TCC TCT TCT TGG TAG CAA CAG CTA CAG GTG TAC ACT CCC AGT CTG TGCTGA CTC AG,SEQ ID NO:131)和反向(CGC CGC CGT ACG TAG GAC CGT CAG CTT GGT,SEQID NO:132)。在以下条件下:95℃/3min;95℃/30sec,60℃/30sec,72℃/30sec,30个循环;72℃/5min,通过使用引物(各10pmol)以各噬菌体DNA(50ng)作为模板,进行PCR,以扩增scFv的VH或VL基因。将PCR产物插入pcDNA3.4载体(Thermo Fisher Scientific),该载体用于通过使用限制性内切酶生产IgG。IgG的重链和轻链蛋白分别在质粒中表达。
将含有编码包括以这种方式制备的scFv的可变区的IgG的轻链和重链的DNA的载体共转化到freestyle293F细胞(ATCC)中,以使轻链和重链在细胞中共表达。将转化的293F细胞在37℃和8%CO2条件下孵育7天以获得上清液。使用醋酸纤维素膜滤膜(孔径为0.22μm,Corning)过滤上清液,并使用CaptivATMPriMAB蛋白A柱(Repligen,美国)进行纯化。使用BCA试剂盒(Pierce,23225)测量获得的抗体浓度,并使用生物分析仪(Agilent2100Bioanalyzer)分析在还原和非还原条件下产生的IgG抗体蛋白。
5-2.免疫细胞迁移和侵袭的抑制效果的确定
证实了5-1中产生的各候选抗体对免疫细胞迁移和侵袭的影响。具体的实验方法如下。将Transwell(Corning#3421-5mm)用明胶(0.5mg/ml)包被,然后将RAW 264.7细胞(1×105细胞/孔)接种到上室中。将含有层粘连蛋白421(1μg/ml)的无血清DMEM(500μl)置于下室中。每种抗体在上室中以100nM的浓度处理。24小时后,将细胞用70%甲醇固定30分钟,然后用50%苏木精染色30分钟。用棉签去除膜上的非迁移细胞后,取出膜并将其装在载玻片上。在高倍率显微镜下观察膜下侧的迁移细胞(图6),并且测量获得的图像中的细胞数并绘图展示(图7)。
此外,通过用层粘连蛋白421(1μg/ml)和抗体(100nM)处理RAW 264.7细胞,孵育细胞24小时并收获细胞。随后,使用ProteoExtractte亚细胞蛋白质组提取试剂盒(Calbiochem),将样品分为膜部分和胞质溶胶部分,然后在KRS上进行蛋白质印迹。具体方法如实施例2所述。
如图6和图7所示,结果证实了本发明的N3单克隆抗体(与KRS N-末端特异性结合的抗体)有效地抑制了依赖LN421的单核细胞/巨噬细胞的迁移。此外,如图8所示,LN421处理增加了单核细胞/巨噬细胞的细胞膜位置的KRS水平,并且证实了通过N3单克隆抗体处理,KRS水平在细胞膜位置特别降低了。另外,如图9所示,证实了当N3抗体结合暴露于细胞外的KRS区域(特别是N-末端区域)时发生内吞作用。这不仅抑制了KRS从细胞质向细胞膜的迁移,而且还表明通过使用与KRS特异结合的物质(特别是,与暴露在细胞外的N-末端特异性结合的物质)主动清除已经迁移到细胞膜的KRS,可以抑制免疫细胞迁移并治疗与免疫细胞迁移相关的疾病。这些结果证实了与KRS的N-末端结合的抗体具有作为免疫细胞迁移/侵袭相关疾病的新型治疗剂的可能性。
5-3.N3抗体测序
使用Omp引物根据Hye young Yang,et.al.,2009,Mol.Cells 27,225-235中描述的方法确定N3scFv(SEQ ID NO:59)和N3IgG抗体(SEQ ID NO:89和SEQ ID NO:91)的序列。使用Bioedit程序确定由此获得的序列的CDR区的序列。抗原结合位点的测序结果如表1所示,且scFv还包括如SEQ ID NO:57所示的接头。
表1.
5-4.N3单克隆抗体的人KRS结合位点的验证
在人KRS的N-末端区域,进行以下实验以确定N3单克隆抗体结合位点。首先,如图10和表2所示,使用作为抗原(表位)的KRS片段肽F1至F5通过表面等离子体共振(SPR)分析了N3抗体的结合能力,该KRS片段F1至F5在序列SEQ ID NO:148的KRS N-末端区域中具有略微不同的位点。使用配备有Series S传感器芯片CM5(GE Healthcare)的Biacore T200(GEHealthcare)在25℃下进行SPR实验。用胺偶联试剂盒(GE Healthcare)将抗体固定在芯片上后,将每种抗原肽溶解在PBS溶液中,然后在156nM-15000nM的范围内稀释两次,并流动60秒。然后使PBS流动300秒。使用软件Biacore T200Evaluation software v2.0(GEHealthcare)分析获得的数据。
表2.
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如图10所示,N3IgG抗体根本不与F5肽结合,且显示的结合能力依次为:F4>F3>F2>F1。所述F4、F3、F2和F1在序列如SEQ ID NO:148所示的人KRS的N-末端共享15至29个氨基酸。由此确认了15-29个氨基酸区域(15-29个氨基酸残基)在N3抗体与人KRS的结合中起着重要作用,并且人KRS的第15-42位氨基酸(在人KRS的第15-29位氨基酸中对应于F4多肽)被认为是N3抗体的主要结合位点。
5-5.N3单克隆抗体在其他动物模型中功效评估的验证
为了证实是否可以通过在小鼠和大鼠中构建疾病动物模型来评估本发明的N3抗体的功效,分析了KRS N-末端序列中跨物种的序列相似性。如图11和表2所示,将KRS片段肽F1至F5用作抗原(表位),该KRS片段肽F1至F5在人(h)、小鼠(m)和大鼠(r)的KRS N-末端区域中具有略微不同的位点。通过表面等离子体共振(SPR)分析N3抗体的结合能力。具体方法如实施例<5-4>中所述。
如图11所示,由于N3抗体在人(h)、小鼠(m)和大鼠(r)中显示出相同的结合模式,因此它根本不结合F5肽。显示的结合能力依次为F4>F3>F2>F1。比较每个物种的F1至F4序列时,它们通常包括KLSKNELKRRLKA序列。得出17-29-氨基酸区域(SEQ ID NO:117)在实施例5-4中确认的KRS的第15-29位氨基酸中的抗体结合中起重要作用。通过该实验得出,N3抗体的种间可能发生交叉反应,这一事实表明小鼠和大鼠模型可用于N3抗体的毒性试验和体内功效试验。
实施例6:体内模型中KRS N-末端特异性结合抗体在免疫细胞迁移相关疾病中的功效验证_体内肺动脉高压模型
特异性结合KRS N-末端的抗体的治疗通过降低细胞膜位置的KRS的水平(通过内吞作用等)来抑制免疫细胞迁移/侵袭,从而产生与实施例4的化合物相同的效果(细胞膜的KRS水平的降低)。因此,显然本发明的KRS N-末端特异性抗体(通常为N3抗体)会表现出与实施例4的化合物的相同适应症(免疫细胞迁移相关的疾病)的疗效,这通过以下实施例表明。
实验方法
1)肺动脉高压(PAH)模型的建立和测试物质的给药
为了在7周龄的SD大鼠(Oriental Bio)中诱导PAH,皮下注射60mpk的MCT(野百合碱)。然后,将大鼠分为四组(每组五只动物进行测试),并施用1mpk的Mock人IgG(ThermoFisher Scientific,阴性对照),1mpk的N3IgG抗体,10mpk的N3IgG抗体和25mpk西地那非(阳性对照)3周。所有抗体均为静脉注射,每周两次注射,西地那非每天口服。
2)血流量和血压测量
三周后,用异氟醚麻醉大鼠,并使用MPVS心血管压力和容量系统(型号名称:MPVSUltra,制造商:Millar Instruments)测量血流量和压力。使用专用导管(Mikro-Tip大鼠压力导管,制造商:Millar Instruments)测量右心室收缩末压(RVESP)、右心室舒张末压、左心室收缩末压、左心室舒张末压。使用血管周围血流探头(Transonic Flow Probes,制造商:Millar Instruments)测量心输出量,并且其实验方法通过与以下文献中公开的方法相同的方法进行:Pacher P,Nagayama T,Mukhopadhyay P,Batkai S,Kass DA。使用压力容量电导导管技术在小鼠和大鼠中测量心脏功能。Nat Protoc 2008;3(9):1422-34。
3)免疫组织化学(IHC)
按照常规步骤将收集的肺固定在PFA(多聚甲醛)中,然后通过洗涤、脱水和清除将其包埋在石蜡中。将大鼠肺组织的石蜡块切成3μm的厚度,制成载玻片。样品首先用二甲苯处理5分钟3次,然后依次用100%乙醇、95%乙醇、90%乙醇和70%乙醇和DW依次处理2分钟,然后用PBS洗涤5分钟。在0.3%H2O2处理后,将样品用PBS洗涤5分钟两次。浸泡在0.01M柠檬酸盐缓冲液中并加热后,样品用PBS-T(0.03%吐温20)洗涤,然后在室温下封闭30分钟(含2%BSA和2%山羊血清的PBS)。将其在4℃下用抗CD68抗体(1:200,ED1克隆,Abcam)染色过夜。用PBS-T洗涤3次,每次5分钟后,将样品用多聚体HRP抗小鼠envision试剂盒(DAKO)在4℃下处理1小时。用PBS-T洗涤3次后,通过DAB底物缓冲液和DAB色原20处理使样品显影。将染色的组织用Mayer's苏木精(Sigma)处理1分钟,然后依次用70%乙醇、90%乙醇、95%乙醇和100%乙醇处理两次,每次2分钟。最后,将组织用二甲苯处理3次,每次5分钟,然后在光学显微镜下观察。
结果
6-1.血压和心输出量变化的验证
PAH(一种疾病,其中免疫细胞侵袭与病理性现象密切相关)模型用N3IgG抗体(1mpk或10mpk)处理了3周(静脉注射,每周两次),然后测量了右心室收缩末压(RVESP)、右心室舒张末压(RVEDP)、左心室收缩末压(LVESP)、左心室舒张末压(LVEDP)和心输出量(CO)。其结果见表3。
表3.
(未测量MCT+Mcok IgG组中一只动物和西地那非治疗组中一只动物的CO,因为它们分别因麻醉和在手术期间死亡)
肺动脉高压因肺动脉变窄而引起右心室压力升高,从而导致右心室衰竭。此外,如果持续性高血压破坏了奖赏机制(reward mechanism),则右心室扩大后会出现右心室扩大。由于室间隔的运动以及左心室舒张末容量和心输出量的减少,从而导致左心室受压(Lee Woo-seok et al.,2007;37:265-270)。结果表明,肺动脉高压主要与右心室有关,但也与左心室的功能有关。
PAH患者显示RVESP升高,在这个实验的PAH动物模型中也证实了这一点。相反,如图12所示,N3抗体(一种特异性结合KRS N末端的抗体)在两种浓度下均显著地降低了RVESP,特别是比阳性对照药物西地那非更好地降低了RVESP。
此外,施用N3抗体(特异性结合KRS N-末端的抗体)后,左心室收缩末压(LVESP)没有降低。相反,如图13所示,LVESP在施用过本发明的抗体的组中显著地增加了。这与通过引起肺动脉扩张以及全身动脉扩张(在用作常规治疗肺动脉高压的西地那非的情况下)而降低全身血压的风险形成对比。也就是说,证实了本发明的抗体与西地那非相比显示出具有对全身动脉压的较低影响的趋势,并且考虑到西地那非的施用可能引起在临床部位出现低血压的风险,这种影响被认为是治疗剂的有利特性。此外,严重的肺动脉高压会导致心脏收缩RV衰竭,并伴有低心输出量和全身性低血压。然而,期望通过缓解肺动脉高压的本发明的N3抗体的治疗来提高心输出量和全身血压,从而使血压正常化。
总之,证实了施用本发明的KRS N-末端结合抗体(特别是N3抗体)降低了现有治疗药物副作用的风险,并显示出PAH症状的缓解和治疗效果。
6-2.超声心动图
在单独MCT的给药组(即不施用测试物质的PAH模型)的3只动物中和在MCT+西地那非给药组的3只动物中观察到D型左心室,这发现表明右心室压力超负荷,而在治疗性抗体给药组中却未观察到。
另外,如下表4所示,各组的重量增加到相似的程度,没有显著差异。也就是说,未观察到表示由于施用治疗性抗体引起的异常体征的现象,包括异常体重减轻。
表4.
6-3.单核细胞/巨噬细胞迁移和浸润程度的验证
通过使用每个实验组的肺组织,对作为单核细胞/巨噬细胞标记物的CD68进行IHC染色。如图14所示,证实了本发明的N3抗体(KRS N-末端结合抗体)治疗组显著地降低了单核细胞/巨噬细胞向肺组织的浸润,这个效果显著优于西地那非的效果。
实施例7:在体内模型中验证KRS-N末端特异性结合抗体对免疫细胞迁移相关疾病的影响_急性肺损伤模型
方法
1)LPS诱发的急性肺损伤模型的构建和测试物质的施用
急性肺损伤的小鼠模型是通过向7周龄的雄性C57BL/6小鼠(duothermal bio)进行气管内注射2.5mg/kg LPS(Sigma)而构建的。为了研究KRS抑制剂对急性肺损伤的影响,首先,分别以1mg/kg或10mg/kg的剂量对C57BL/6小鼠静脉注射N3IgG抗体,然后24小时后,气管内注射2.5mg/kg LPS。LPS注射24小时后,安乐死每只小鼠以收集和分析肺组织和BALF(支气管肺泡灌洗液)。
2)支气管肺泡灌洗液(BALF)中的免疫细胞计数
收集通过用PBS洗涤肺而获得的BALF,并通过在4℃下以800x g离心10分钟来收集沉淀。悬浮细胞后,使用RBC裂解缓冲液(eBioscience目录号00-4333-57)洗去红细胞。用PBS终止反应后,洗涤两次,并悬浮在400μl的PBS中以通过血球计数板测量细胞数并通过Hema3染色测量中性粒细胞数。
3)FACS对肺组织免疫细胞的影响
收集肺组织,并使用温和的MACS Octo Dissociator(MACS Miltenyi Biotec,订购号130-095-937)在37℃旋转45分钟以压碎组织。使用细胞过滤器(40μm)过滤后,在室温下以1500rpm离心5分钟。收集沉淀并使用RBC裂解缓冲液(eBioscience目录号00-4333-57)洗去红细胞。收集细胞并将其悬浮在FACS缓冲液(含1%NaN3和3%FBS的PBS)中,将50μl置于试管中,与等量的抗体混合物充分混合,并在4℃下通过阻挡光线染色1小时。使用FITC大鼠抗CD11b(BD Pharmingen)和PE大鼠抗小鼠F4/80(BD Pharmingen)抗体来分析向肺移动的间质巨噬细胞(IM)。使用FACS缓冲液以400×g洗涤两次(每次5分钟)后,通过Navios流式细胞仪(Beckman)进行分析。
4)肺组织的Masson三色染色
将肺组织以原始方式包埋在石蜡中,然后切出。然后,用DW洗涤用二甲苯去除了石蜡的组织玻片,然后在56-60℃下用布安液(Bouin Fluid)处理1小时。用Weigert氏苏木精溶液染色10分钟后,洗涤组织玻片。再次用Biebrich绯红酸品红溶液(Biebrich scarlet-acid fuchsin solution)染色10-15分钟后,洗涤玻片。将磷钼酸盐-磷钨酸溶液在玻片上处理10-15分钟,然后将玻片转移到苯胺蓝溶液中并染色5-10分钟。洗涤后,将玻片用1%乙酸溶液处理2-5分钟。洗涤和脱水后,将玻片用二甲苯处理并固定。
结果
7-1.对支气管肺泡灌洗液(BALF)中免疫细胞迁移的抑制效果的验证
如图15所示,证实了在通过LPS处理引起的急性肺损伤小鼠中,BALF中的免疫细胞总数增加了,其通过N3抗体(KRS N-末端结合抗体)处理浓度依懒性地降低了。特别地,如图16所示,证实了在通过LPS处理患有急性肺损伤的小鼠中,中性粒细胞增加了,并且N3抗体(KRS N-末端结合抗体)治疗降低了这些中性粒细胞的水平。结果证实了通过处理KRS N端特异性结合的抗体,显著地抑制了免疫细胞(特别是中性粒细胞)浸润到BALF的肺。
7-2.抗体对肺组织中免疫细胞迁移的抑制效果的验证
图17和18显示了由于急性肺损伤而迁移至肺组织的巨噬细胞的FACS分析结果。间质巨噬细胞(IM)是CD11b+/F4/80+细胞,它们是迁移的巨噬细胞,它们不驻留在肺中,而是在某些情况下会迁移到肺中。LPS处理增加了IM向肺的浸润,而N3抗体处理以依赖于浓度的方式减少了IM向肺的迁移。由此证实,通过特异性结合KRS N-末端的抗体(通常为N3抗体)的治疗,抑制了巨噬细胞/单核细胞等免疫细胞向肺组织的迁移和侵袭。免疫细胞(例如巨噬细胞/单核细胞)的过度迁移和侵袭是组织纤维化疾病中的一个重要病理现象。通过对急性肺损伤模型的肺组织的Masson三色染色的观察(图19),证实了肺组织中的相当大的纤维化进展。相反,证实了N3抗体(特异性结合KRS N-末端的抗体)的治疗抑制了这种纤维化。
实施例8:对KRS的N-末端具有增加的亲和力的N3修饰的抗体的构建
本发明人试图通过修饰N3抗体来获得对KRS的N-末端区域具有更好亲和力的抗体,以生产表现更好性能的作为治疗性抗体的抗体。因此,通过以下一系列过程改善了N3抗体的轻链可变区和重链可变区。
8-1.基于N3抗体的序列变体中的scFab文库(酵母细胞表面表达文库)的构建_初级文库
使用同源建模方法,可以预测N3抗体的大致结构,并由此将随机突变引入预测在抗原结合中起重要作用的CDR区域,以构建文库。具体地,在基于重链可变区的文库中,使用了NNK,该NNK为可包含重链CDR2或CDR3中的残基的所有20个氨基酸序列的简并密码子。在轻链可变区文库中,使用了NNK,该NNK为该密码子可以包含N3抗体轻链CDR2或CDR3中的残基的所有20个氨基酸序列的简并密码子。使用PCR技术扩增编码设计的文库的DNA,然后使用乙醇沉淀法浓缩。
然后,参照以下文献中描述的方法,构建在酵母表面表达的包含轻链可变区各种变化的scFab文库:Baek DS and Kim YS,Construction of a large synthetic humanFab antibody library on yeast cell surface by optimized yeast mating,JMicrobiol Biotechnol.2014Mar 28;24(3):408-20。
简而言之,用NheI和MluI限制性内切酶处理在C-末端表达aga2蛋白以进行同源重组的酵母表面表达载体(C-aga2:pYDS),用琼脂糖凝胶提取法纯化,并用乙醇沉淀法浓缩。
通过电穿孔,将12μg每种文库编码DNA中的、限制酶处理过的4μg载体通过电穿孔转化到酵母EBY100(酵母用于表面表达)中,并通过测量选择培养基[经连续稀释的SD-CAA(20g/L葡萄糖,6.7g/L不含氨基酸的酵母氮碱、5.4g/L Na2HPO4、8.6g/L NaH2PO4、5g/L酪蛋白氨基酸)]中生长的菌落数来确定文库大小,。
8-2.对KRS(残基1-72氨基酸)肽具有改善的亲和力的scFab筛选_第一文库的选择
使用缀合GST的KRS(残基1-72氨基酸,N-末端)肽作为抗原,为实施例8-1中构建的文库选择具有增加的亲和力的Fab。
具体地,使用用于初级FACS文库筛选的SG-CAA培养基(20g/L半乳糖,6.7g/L不含氨基酸的酵母氮碱,5.4g/L Na2HPO4、8.6g/L NaH2PO4,5g/L酪蛋白氨基酸),使10nM缀合GST的KRS肽(残基1-72氨基酸,纯化状态)在室温下与诱导实施例8-1中的细胞表面的scFab文库表达的酵母反应1小时。然后,使缀合GST的KRS(残基1-72)肽和表达文库的酵母与缀合PE的链霉亲和素-R-藻红蛋白缀合物(SA-PE)在4℃下反应20分钟,并通过FACS悬浮(荧光激活细胞分选术,FACS Caliber;BD biosciences)。随后,用缀合1nM GST的KRS(残基1-72)肽进行第二次FACS筛选,并且用缀合0.5nM GST的KRS(残基1-72)肽进行第三次FACS筛选。
通过使用FACS的筛选过程,选择了与N3抗体相比对KRS(残基1-72)肽具有高亲和力的克隆。以此,通过分析单个克隆的结合能力,选择了对KRS(残基1-72)肽具有高亲和力和特异性的三个优良克隆(N3-1、N3-3、N3-4),以及另一个轻链可变区和重链可变区彼此结合以构建另一个独特的克隆N3-5。还证实了N3-5克隆对KRS(残基1-72)肽具有极好的亲和力和特异性,且最后筛选并选择了总共4个独特的(磁性激活细胞分选术)克隆N3-1,N3-3,N3-4,N3-5。
表5显示出四个单个克隆N3-1、N3-3、N3-4和N3-5的轻链可变区和重链可变区的CDR序列的较高的与KRS(残基1-72)肽的结合能力。表6显示了重链可变区和轻链可变区的完整序列。
表5.
表6
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此外,使用ELISA法再次证实了所选的单个克隆N3-1、N3-3、N3-4和N3-5对KRS N-末端的亲和力。在该实验中,测试了转化到IgG抗体的克隆。转化到IgG抗体参见以上实施例5-1的方法。
具体地,在96孔EIA/RIA板(COSTAR Corning)中处理KRS的N-末端区域(残基1-72)肽,并在25℃下结合1小时,然后用PBS(pH 7.4,137mM NaCl,12mM磷酸盐,2.7mM KCl)(SIGMA)洗涤3次,每次10分钟。然后,将4%BSA PBS(4%牛血清白蛋白,pH7.4,137mM NaCl,12mM磷酸盐,2.7mM KCl)(SIGMA)处理1小时,然后用PBS洗涤3次,每次10分钟。然后,分别处理和结合IgG型的N3抗体N3-1抗体、N3-3抗体、N3-4抗体和N3-5抗体,然后用0.1%PBST洗涤3次,每次10分钟。使用缀合辣根过氧化物酶的抗人mAb(SIGMA)作为标记的抗体。然后,使其与TMB(3,3',5,5'-四甲基联苯胺)(Sigma)反应,并在450nm吸光度下进行测量以进行定量。
如图20和21所示,结果证实了与N3抗体相比,作为改进的抗体的N3-1、N3-3、N3-4和N3-5抗体的亲和力显著地增加了。发现了所有克隆均不结合作为阴性对照的GST或NRP1-b1b2。在突变体抗体N3-1、N3-3、N3-4和N3-5内,KRS结合能力没有显著差异(见图21)。
8-3.N3抗体与初级改进的抗体之间的亲和力比较及其对细胞迁移的抑制效果的验证
KRS与改进的抗体N3-1、N3-3、N3-4和N3-5之间的结合能力没有显著差异。因此,使用在实施例8-2中选择的改进的抗体中的代表性的N3-1IgG抗体,更具体地比较了亲本抗体N3抗体与KRS N-末端的亲和力。使用KRS片段(1-207氨基酸)纯化蛋白作为抗原,通过表面等离子体共振(SPR)分析了N3抗体与N3-1抗体(IgG)之间的结合能力。使用配备有Series S传感器芯片CM5(GE Healthcare)的Biacore T200(GE Healthcare)在25℃下进行SPR实验。使用胺偶联试剂盒(GE Healthcare)将抗体固定在芯片上后,将抗原在4.8nM-1250nM的PBS溶液中稀释4倍,并流动60秒。然后,使PBS流动300秒。使用软件Biacore T200Evaluationsoftwarev 2.0(GE Healthcare)分析获得的数据。
如图22所示,结果显示,N3-1抗体的KD值测量得31nM,这表明与N3抗体相比,与KRS蛋白的结合能力提高了。此外,通过抗体处理引起(参见上述实施例)对细胞迁移的抑制效应的结果,证实了与N3抗体相比,N3-1抗体显著地抑制了细胞迁移。
8-4.在基于N3抗体的序列变体(酵母细胞表面表达文库)构建scFab文库_次级文库
实施例8-2中衍生的靶向KRS N-末端的N3-1、N3-3、N3-4和N3-5抗体具有与KRS相似的亲和力,且如实施例8-3中N3-1抗体的结果所示,确定了具有约31nM的亲和力。为了通过增加它们对KRS的亲和力(特别是N-末端区域)来获得更有效的抗体,极大地改善了重链可变区。
轻链可变区序列确定用N3-3抗体的序列(SEQ ID NO:51),并且用重链可变区的不同的可变序列构建文库。首先,使用同源建模方法预测N3-3抗体的近似建模结构,然后,将随机突变引入被预测在抗原结合中起重要作用的CDR。具体地,将能够包含全部20个氨基酸序列的简并密码子NNK用作N3-3抗体的重链可变区的CDR2和CDR3的残基,并以与实施例8-1所示的相同的方式与构建scFab文库。
8-5.筛选具有改善的KRS(残基1-72)亲和力的ScFab_次级文库的筛选
使用缀合GST的KRS(残基1-72)肽作为抗原,在实施例8-4中构建的文库中选择具有增加的亲和力的Fab。由于确定了N3-3和N3-1的亲和力几乎相同并且序列几乎相似,因此使用N3-1进行了对比实验。
首先,用实施例8-4中构建的表达文库的酵母处理缀合GTP的KRS(残基1-72)肽。然后,使表达与GTP偶联的KRS(第1-72位)肽结合的文库的酵母与链霉亲和素微珠TM(MiltenyiBiotec)在4℃下反应20分钟,然后,使用磁激活细胞分选术(MACS),将使表达对KRS(1-72氨基酸)肽具有高亲和力的scFab的酵母悬浮。通过MACS选择的表达文库的酵母在SG-CAA(20g/L半乳糖,6.7g/L不含氨基酸的酵母氮碱,5.4g/L Na2HPO4、8.6g/L NaH2PO4、5g/L酪蛋白氨基酸)培养基中培养以诱导文库表达。随后,以与实施例8-2相同的方式,使用FACS依次进行筛选。简而言之,第一次FACS筛选是用结合了10nM GST的KRS(1-72)肽进行的,第二次FACS筛选是用结合了1nM GST的KRS(1-72)肽进行的,第三次FACS筛选是用结合了0.5nMGST的KRS(1-72)肽进行的,以及第四次FACS筛选是用结合了0.1nM GST的KRS(1-72)肽进行的。
通过使用磁激活细胞分选术(MACS)和荧光激活细胞分选术(FACS)的筛选过程,选择了与N3-1抗体(显示出与N3-3抗体相似的亲和力)相比,对KRS(1-72氨基酸)肽具有高亲和力的克隆(高亲和力取决于重链可变区(VH))。以这种方式,通过对单个克隆的结合能力分析,选择了对KRS(残基1-72)肽具有高亲和力和特异性的四个优良克隆N3-6、N3-7、N3-8和N3-9。
表7显示了与KRS(1-72氨基酸)肽具有高结合能力的四个单个的克隆N3-6、N3-7、N3-8和N3-9的轻链可变区和重链可变区的CDR序列,表8显示了重链可变区序列和轻链可变区的完整序列。
表7.
表8.
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此外,使用ELISA法再次确认了所选的单个克隆N3-6、N3-7、N3-8和N3-9对KRS N-末端的亲和力。在该实验中,将克隆转化到IgG抗体后,将其用于ELISA实验,该方法参见以上实施例5-1。
如图23所示,结果证实了,与N3-1抗体(与N3-3类似)相比,改进的抗体N3-6、N3-7、N3-8和N3-9的亲和性提高。对于用作阴性对照的NRP1-b1b2,发现没有克隆被结合。
此外,通过作为改进的抗体的N3-6抗体、N3-7抗体、N3-8抗体和N3-9抗体(IgG)的处理而抑制细胞迁移的结果(方法参见上述实施例)证实了与N3-1抗体相比,改进的抗体显著地抑制细胞迁移。N3-6抗体、N3-7抗体、N3-8抗体和N3-9抗体对细胞迁移的抑制效应没有显著差异。
8-6.表位和改进的抗体N3-6、N3-7、N3-8和N3-9的亲和力的验证
使用KRS表位肽F4(EPKLSKNELKRRLKAEKKVAEKEAKQKE:SEQ ID NO:136)作为抗原表位,通过表面等离子体共振(SPR)分析N3抗体、N3-6抗体、N3-7抗体、N3-8抗体和N3-9抗体的结合能力。以与实施例5-4相同的方式进行SPR实验。将表位在PBS溶液中稀释,并在15.7nM-4000nM的范围内稀释2倍,并使其流动90秒。然后,使PBS流动2400秒。使用软件BiacoreT200Evaluation software v2.0(GE Healthcare)分析获得的数据。
如图24所示,结果还证实了,改进的抗体N3-6抗体、N3-7抗体、N3-8抗体和N3-9抗体具有与N3抗体相似的主要结合位点(参见实施例5-4)。此外,使用其中KRS表位肽F4的组成氨基酸(SEQ ID NO:136)分别被丙氨酸(A)取代的肽,进行ELISA以确定对于抗体-表位结合重要的残基。结果显示,可在KRS表位肽F4中确定对结合各抗体重要的残基。
此外,如图24所示,N3-8抗体的KD(KD=0.8613nM)表现优良,N3-9抗体的KD(KD=2.842nM)和N3-6抗体的KD(KD=4.114nM)相似,并且N3-7抗体的KD值(KD=18.77nM)最大。N3-6抗体的解离时间比N3-7和N3-9的解离时间长,并且显示出具有更长时间结合的传感图。
8-7.基于N3-8抗体生产高产和稳定性的改良抗体(N3-8抗体序列改进)
在以上实施例中,证实了N3-8抗体对KRS具有最佳的亲和力(特别是N-末端)。因此,为了确定诸如N3-8抗体的生产率和稳定性的特性并使这些特性更好,通过诱导序列中预期影响N3-8抗体序列稳定性的突变来生产N3-8衍生物。
结果得到两个重链序列(SEQ ID NO:45,SEQ ID NO:47),其中突变被引入到N3-8抗体的重链可变区中。此外,获得了三个轻链序列(SEQ ID NO:51,SEQ ID NO:53,SEQ IDNO:55),其中突变被引入到轻链可变区中。下表9和10显示了根据重链和轻链序列的组合的七种类型的N3-8衍生物的序列,它们保持了N3-8抗体的亲和力特性。
表9.
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表10.
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表11显示了上述实施例中使用的全部IgG抗体的重链(HC)和轻链(LC)序列。
表11.
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8-8.N3-8衍生物的生产率和稳定性的测量(Tm测量)
使用表达实施例8-7中获得的N3-8衍生抗体的轻链和重链的质粒,通过瞬时转染方法表达和纯化每种抗体蛋白。在摇瓶中悬浮于无血清FreeStyle 293表达培养基(Invitrogen)中的HEK293-F细胞(Invitrogen)中,转染质粒和聚乙烯亚胺(Polyethylenimine,Polyscience)。
具体地,在转染到200mL摇瓶中的过程中,将HEK293-F细胞以2×106个细胞/ml的密度接种在100ml培养基中,并在8%CO2、150rpm和37℃温度条件下培养。通过以1∶1或1∶2比例的重链:轻链DNA处理它们,转染适于生产每种单克隆抗体的重链质粒和轻链质粒(在10ml FreeStyle 293表达培养基中混合处理)。首先,当以1:1的比例使用重链:轻链DNA时,将总共125μg重链,125μg轻链和250μg(2.5μg/ml)与10ml用PEI 750μg(7.5μg/ml)稀释过的培养基在室温下混合。反应进行10分钟。在比例为1:2的情况下,轻链DNA的浓度加倍。然后,将混合培养基用预先等分100ml的HEK293-F细胞处理,并在150rpm和37℃,8%CO2条件下孵育4小时。然后加入剩余的100ml FreeStyle 293表达培养基并培养6天。
然后,将细胞培养液转移至50ml试管中,并以3000rpm离心5分钟。然后从收集的细胞培养上清液中纯化蛋白质。将上清液加到蛋白A Sepharose柱上,然后用PBS(pH7.4)洗涤。用0.1M甘氨酸缓冲液在pH 3.0下洗脱抗体后,立即用1M Tris缓冲液中和样品。通过透析法通过与PBS(pH 7.4)交换缓冲液来浓缩洗脱的抗体部分。使用在280nm波长的吸光度和吸收系数对纯化的蛋白质进行定量。
此外,使用100μl浓度为1mg/ml的纯化抗体测量抗体的热稳定性。使用QuantStudio3实时PCR设备(Thermofisher)进行4次蛋白质热转移染料试剂盒(Protein thermalshift Dye kit)(Thermofisher)。
如下表12所示,结果证实了所测试的所有N3-8衍生物抗体的产量均高于N3-8抗体的产量。此外,如表12所示,获得了Tm值,并且根据抗体Tm1-Tm2观察到热转化不同,然而却发现在所有N3-8衍生抗体中Tm值均增加。
由此证实了N3-8抗体衍生物比N3-8抗体具有更高的产量和改善的热稳定性。
表12.
(在上表中,“–”表示与Tm1相同的值)
实施例9:证实N3改进的抗体的疗效增加
对于在以上实施例中生产的N3改进的抗体,证实了针对免疫细胞迁移相关疾病(即使在体内),其疗效是否显著。使用肺动脉高压模型作为疾病模型,且N3-6抗体和N3-8抗体通常被用作改进的抗体。
为了在7周龄的SD大鼠(Orient Bio)中诱发肺动脉高压,皮下注射60mpk的MCT(野百合碱)。诱导疾病2周后,分为4组(每组5个实验),分别施用PBS、1mpk N3抗体1mpk、1mpkN3-6抗体和1mpk N3-8抗体,持续3周。所有抗体静脉内注射,每周两次。以与上述实施例6所述相同的方式,对每个实验组进行血流量和血压的测量以及免疫组化染色(IHC)。此外,用Masson三色染色法对每个实验组的肺组织进行染色以测量肺纤维化程度,且以与实施例7所述相同的方式进行Masson三色染色法。
在仅经MCT处理的动物的肺中,观察到肺血管内皮炎、小动脉平滑肌增生、内膜增生和血管闭塞。因此,证实了较好地生产了肺动脉高压疾病的动物模型。
通过本发明的抗体的处理,肺动脉高压表现出的现象显著地减少了。具体地,如图25所示,证实了通过本发明的N3抗体和N3改进的抗体的处理,肺动脉高压(仅MCT治疗组)中增加的RVESP显著地降低了,特别是N3-8抗体具有优良的RVESP降低效果。
图26显示了每个对照组和实验组的肺组织的IHC染色结果。证实了在肺动脉高压(仅MCT治疗组)动物的肺中观察到CD68+单核细胞/巨噬细胞染色强烈,而在N3抗体治疗组或N3-6抗体治疗组中,染色程度减弱,以及在N3-8抗体治疗组中,染色强度显著地降低至与阴性对照相似的水平。这意味着通过本发明的抗体处理抑制了组织中单核细胞/巨噬细胞的浸润,且特别地N3-8抗体表示出优良的效果。
此外,如图27所示,证实了在肺动脉高压(仅MCT治疗组)动物的肺组织中观察到极高的肺纤维化水平,而在N3抗体治疗组或N3-6抗体治疗组中,纤维化症状水平明显减弱,在N3-8抗体治疗组中,纤维化程度降低至与阴性对照相似的水平。这意味着通过本发明的抗体处理纤维化降低了,特别地N3-8抗体表示出优良的效果。
工业适用性
本发明涉及一种与赖氨酰-tRNA合成酶的N-末端特异性结合的抗体的新用途,且更具体地,涉及一种用于预防和治疗免疫细胞迁移相关疾病的、含作为有效成分的抗体或其功能片段的药物组合物,所述抗体与赖氨酰-tRNA合成酶(KRS)的N-末端区域中含序列SEQ ID NO:117的表位特异性结合。本发明提供的KRS N-末端特异性抗体可以调节免疫细胞迁移,从而可非常实用地用于预防、缓解和治疗免疫细胞迁移相关疾病并因此具有极高的工业适用性。
<110> 塞梅蒂股份有限公司
<120> 用于预防或治疗细胞迁移相关疾病的含作为有效成分的
与赖氨酰-tRNA合成酶N-末端特异性结合的抗体的药物组合物
<130> P20111064WP
<140> CN2018800743725
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50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 32
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH
<400> 32
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcttatg ataatggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 33
<211> 110
<212> PRT
<213> 人工序列
<220>
<223> N3 VL
<400> 33
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Trp Asp Asp Ser Leu
85 90 95
Ser Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 34
<211> 330
<212> DNA
<213> 人工序列
<220>
<223> N3 VL
<400> 34
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat gataatagta atcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccgg 240
tccgaggatg aggctgatta ttactgtgct tcttgggatg atagcctgag tgcttatgtc 300
ttcggcggag gcaccaagct gacggtccta 330
<210> 35
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> N3 VH 突变体 1
<400> 35
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Val Ile Ser Ser Asp Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 36
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH 突变体 1
<400> 36
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagtg atctcttctg atggtggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 37
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> N3 VH 突变体 2
<400> 37
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Gln Met Gly Arg Val Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 38
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH 突变体 2
<400> 38
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcgccgc agatgggtcg ggtgtattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 39
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> N3 VH 突变体 3
<400> 39
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Asp Pro Leu Gly Gly Asn Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 40
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH 突变体 3
<400> 40
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atcgatccgt tggggggtaa tatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 41
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> N3 VH 突变体 4
<400> 41
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 42
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH 突变体 4
<400> 42
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 43
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> N3 VH 突变体 5
<400> 43
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Gly Ala Asp Gly Gly Pro Ser Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 44
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH 突变体 5
<400> 44
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atcggggctg atgggggtcc gtcttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 45
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> N3 VH 突变体 6
<400> 45
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 46
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH 突变体 6
<400> 46
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 47
<211> 116
<212> PRT
<213> 人工序列
<220>
<223> N3 VH 突变体 7
<400> 47
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Leu Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser
115
<210> 48
<211> 348
<212> DNA
<213> 人工序列
<220>
<223> N3 VH 突变体 7
<400> 48
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagactggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctca 348
<210> 49
<211> 110
<212> PRT
<213> 人工序列
<220>
<223> N3 VL 突变体 1
<400> 49
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 50
<211> 330
<212> DNA
<213> 人工序列
<220>
<223> N3 VL 突变体 1
<400> 50
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat gataatagta atcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgtgct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta 330
<210> 51
<211> 110
<212> PRT
<213> 人工序列
<220>
<223> N3 VL 突变体 2
<400> 51
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 52
<211> 330
<212> DNA
<213> 人工序列
<220>
<223> N3 VL 突变体 2
<400> 52
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat gataatagta atcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgttct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta 330
<210> 53
<211> 110
<212> PRT
<213> 人工序列
<220>
<223> N3 VL 突变体 3
<400> 53
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 54
<211> 330
<212> DNA
<213> 人工序列
<220>
<223> N3 VL 突变体 3
<400> 54
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat agtaataatc agcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgttct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta 330
<210> 55
<211> 110
<212> PRT
<213> 人工序列
<220>
<223> N3 VL 突变体 4
<400> 55
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 110
<210> 56
<211> 330
<212> DNA
<213> 人工序列
<220>
<223> N3 VL 突变体 4
<400> 56
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat cggaataatc agcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgttct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta 330
<210> 57
<211> 15
<212> PRT
<213> 人工序列
<220>
<223> scFv 接头
<400> 57
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 58
<211> 45
<212> DNA
<213> 人工序列
<220>
<223> scFv 接头
<400> 58
ggtggaggcg gttcaggcgg aggtggatcc ggcggtggcg gatcg 45
<210> 59
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3 scFv (VH+接头+VL)
<400> 59
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Tyr Asp Asn Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Arg Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Trp Asp
210 215 220
Asp Ser Leu Ser Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 60
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3 scFv (VH+接头+VL)
<400> 60
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcttatg ataatggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cggtccgagg atgaggctga ttattactgt 660
gcttcttggg atgatagcct gagtgcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 61
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-1 scFv (VH+接头+VL)
<400> 61
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Tyr Asp Asn Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 62
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-1 scFv (VH+接头+VL)
<400> 62
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcttatg ataatggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
gcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 63
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-3 scFv (VH+接头+VL)
<400> 63
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Tyr Asp Asn Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 64
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-3 scFv (VH+接头+VL)
<400> 64
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcttatg ataatggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 65
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-4 scFv (VH+接头+VL)
<400> 65
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Val Ile Ser Ser Asp Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 66
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-4 scFv (VH+接头+VL)
<400> 66
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagtg atctcttctg atggtggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
gcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 67
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-5 scFv (VH+接头+VL)
<400> 67
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Val Ile Ser Ser Asp Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 68
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-5 scFv (VH+接头+VL)
<400> 68
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagtg atctcttctg atggtggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 69
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-6 scFv (VH+接头+VL)
<400> 69
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Gln Met Gly Arg Val Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 70
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-6 scFv (VH+接头+VL)
<400> 70
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcgccgc agatgggtcg ggtgtattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 71
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-7 scFv (VH+接头+VL)
<400> 71
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Asp Pro Leu Gly Gly Asn Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 72
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-7 scFv (VH+接头+VL)
<400> 72
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atcgatccgt tggggggtaa tatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 73
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-8 scFv, N3-8-7 scFv (VH+接头+VL)
<400> 73
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 74
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-8 scFv, N3-8-7 scFv (VH+接头+VL)
<400> 74
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 75
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-9 scFv (VH+接头+VL)
<400> 75
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Gly Ala Asp Gly Gly Pro Ser Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 76
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-9 scFv (VH+接头+VL)
<400> 76
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atcggggctg atgggggtcc gtcttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 77
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-8-1 scFv (VH+接头+VL)
<400> 77
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 78
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-8-1 scFv (VH+接头+VL)
<400> 78
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 79
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-8-2 scFv (VH+接头+VL)
<400> 79
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 80
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-8-2 scFv (VH+接头+VL)
<400> 80
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatagtaata atcagcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 81
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-8-3 scFv (VH+接头+VL)
<400> 81
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 82
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-8-3 scFv (VH+接头+VL)
<400> 82
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatcggaata atcagcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 83
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-8-4 scFv (VH+接头+VL)
<400> 83
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Leu Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 84
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-8-4 scFv (VH+接头+VL)
<400> 84
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagactggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatgataata gtaatcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 85
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-8-5 scFv (VH+接头+VL)
<400> 85
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Leu Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 86
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-8-5 scFv (VH+接头+VL)
<400> 86
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagactggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatagtaata atcagcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 87
<211> 241
<212> PRT
<213> 人工序列
<220>
<223> N3-8-6 scFv (VH+接头+VL)
<400> 87
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Leu Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly
115 120 125
Gly Gly Ser Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr
130 135 140
Pro Gly Gln Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile
145 150 155 160
Gly Ser Asn Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
165 170 175
Lys Leu Leu Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp
180 185 190
Arg Phe Ser Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
195 200 205
Gly Leu Gln Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser
210 215 220
Asp Glu Leu Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val
225 230 235 240
Leu
<210> 88
<211> 723
<212> DNA
<213> 人工序列
<220>
<223> N3-8-6 scFv (VH+接头+VL)
<400> 88
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagactggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagg tggaggcggt 360
tcaggcggag gtggatccgg cggtggcgga tcgcagtctg tgctgactca gccaccctca 420
gcgtctggga cccccgggca gagggtcacc atctcttgta ctggctcttc atctaatatt 480
ggcagtaatt atgtcacctg gtaccagcag ctcccaggaa cggcccccaa actcctcatc 540
tatcggaata atcagcggcc aagcggggtc cctgaccgat tctctggctc caagtctggc 600
acctcagcct ccctggccat cagtgggctc cagtccgagg atgaggctga ttattactgt 660
tcttctttta gtgatgagtt gggggcttat gtcttcggcg gaggcaccaa gctgacggtc 720
cta 723
<210> 89
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> N3 IgG 重链
<400> 89
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Tyr Asp Asn Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 90
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> N3 IgG 重链
<400> 90
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcttatg ataatggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 91
<211> 217
<212> PRT
<213> 人工序列
<220>
<223> N3 IgG 轻链
<400> 91
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Trp Asp Asp Ser Leu
85 90 95
Ser Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr
100 105 110
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
115 120 125
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
130 135 140
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
145 150 155 160
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
165 170 175
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
180 185 190
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
195 200 205
Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 92
<211> 651
<212> DNA
<213> 人工序列
<220>
<223> N3 IgG 轻链
<400> 92
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat gataatagta atcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccgg 240
tccgaggatg aggctgatta ttactgtgct tcttgggatg atagcctgag tgcttatgtc 300
ttcggcggag gcaccaagct gacggtccta cgtacggtgg ctgcaccatc tgtcttcatc 360
ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg cctgctgaat 420
aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct ccaatcgggt 480
aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag cctcagcagc 540
accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg cgaagtcacc 600
catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg t 651
<210> 93
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> IgG 重链 1
<400> 93
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Val Ile Ser Ser Asp Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 94
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> IgG 重链 1
<400> 94
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagtg atctcttctg atggtggtaa tacatattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 95
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> IgG 重链 2
<400> 95
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Gln Met Gly Arg Val Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 96
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> IgG 重链 2
<400> 96
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctcgccgc agatgggtcg ggtgtattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 97
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> IgG 重链 3
<400> 97
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Asp Pro Leu Gly Gly Asn Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 98
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> IgG 重链 3
<400> 98
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atcgatccgt tggggggtaa tatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 99
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> IgG 重链 4
<400> 99
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 100
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> IgG 重链 4
<400> 100
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 101
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> IgG 重链 5
<400> 101
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Gly Ala Asp Gly Gly Pro Ser Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Ser
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 102
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> IgG 重链 5
<400> 102
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atcggggctg atgggggtcc gtcttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactctgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 103
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> IgG 重链 6
<400> 103
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Met Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 104
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> IgG 重链 6
<400> 104
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagaatggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 105
<211> 446
<212> PRT
<213> 人工序列
<220>
<223> IgG 重链 7
<400> 105
Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30
Asp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Ala Ile Ser Pro Tyr Ser Gly Arg Ile Tyr Tyr Ala Asp Ser Val
50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95
Ala Arg Leu Ala Leu Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
100 105 110
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
115 120 125
Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu
130 135 140
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly
145 150 155 160
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser
165 170 175
Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu
180 185 190
Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr
195 200 205
Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr
210 215 220
Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe
225 230 235 240
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro
245 250 255
Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
260 265 270
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
275 280 285
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val
290 295 300
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys
305 310 315 320
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser
325 330 335
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro
340 345 350
Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
355 360 365
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly
370 375 380
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
385 390 395 400
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
405 410 415
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His
420 425 430
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
435 440 445
<210> 106
<211> 1338
<212> DNA
<213> 人工序列
<220>
<223> IgG 重链 7
<400> 106
gaggtgcagc tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60
tcctgtgcag cctctggatt cacctttagc agttatgata tgagctgggt ccgccaggct 120
ccagggaagg ggctggagtg ggtctcagcg atctctccgt attcgggtag gatttattac 180
gctgattctg taaaaggtcg gttcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggccgtgt attactgtgc gagactggcg 300
cttgatttcg actactgggg ccagggtaca ctggtcaccg tgagctcagc ctccaccaag 360
ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420
ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480
gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540
ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600
gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660
aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 720
ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780
gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840
gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900
gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960
aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 1020
cagccccgag aaccacaggt gtataccctg cccccatccc gggatgagct gaccaagaac 1080
caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140
gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200
ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260
gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320
tccctgtccc cgggtaaa 1338
<210> 107
<211> 217
<212> PRT
<213> 人工序列
<220>
<223> IgG 轻链 1
<400> 107
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr
100 105 110
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
115 120 125
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
130 135 140
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
145 150 155 160
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
165 170 175
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
180 185 190
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
195 200 205
Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 108
<211> 651
<212> DNA
<213> 人工序列
<220>
<223> IgG 轻链 1
<400> 108
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat gataatagta atcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgtgct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta cgtacggtgg ctgcaccatc tgtcttcatc 360
ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg cctgctgaat 420
aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct ccaatcgggt 480
aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag cctcagcagc 540
accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg cgaagtcacc 600
catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg t 651
<210> 109
<211> 217
<212> PRT
<213> 人工序列
<220>
<223> IgG 轻链 2
<400> 109
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Arg Thr
100 105 110
Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
115 120 125
Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro
130 135 140
Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
145 150 155 160
Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr
165 170 175
Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
180 185 190
Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
195 200 205
Thr Lys Ser Phe Asn Arg Gly Glu Cys
210 215
<210> 110
<211> 651
<212> DNA
<213> 人工序列
<220>
<223> IgG 轻链 2
<400> 110
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat gataatagta atcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgttct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta cgtacggtgg ctgcaccatc tgtcttcatc 360
ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg cctgctgaat 420
aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct ccaatcgggt 480
aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag cctcagcagc 540
accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg cgaagtcacc 600
catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg t 651
<210> 111
<211> 216
<212> PRT
<213> 人工序列
<220>
<223> IgG 轻链 3
<400> 111
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Asp Asn Ser Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln
100 105 110
Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu
115 120 125
Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr
130 135 140
Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys
145 150 155 160
Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr
165 170 175
Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His
180 185 190
Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys
195 200 205
Thr Val Ala Pro Thr Glu Cys Ser
210 215
<210> 112
<211> 648
<212> DNA
<213> 人工序列
<220>
<223> IgG 轻链 3
<400> 112
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat gataatagta atcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgttct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta ggtcagccca aggctgcccc ctcggtcacg 360
ctcttcccac cctcctctga ggagcttcaa gccaacaagg ccacactggt gtgtctcata 420
agtgacttct acccgggagc cgtgacagtg gcctggaagg cagatagcag ccccgtcaag 480
gcgggagtgg agaccaccac accctccaaa caaagcaaca acaagtacgc ggccagcagc 540
tacctgagcc tgacgcctga gcagtggaag tcccacagaa gctacagctg ccaggtcacg 600
catgaaggga gcaccgtgga gaagacagtg gcccctacag aatgctct 648
<210> 113
<211> 216
<212> PRT
<213> 人工序列
<220>
<223> IgG 轻链 4
<400> 113
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln
100 105 110
Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu
115 120 125
Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr
130 135 140
Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys
145 150 155 160
Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr
165 170 175
Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His
180 185 190
Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys
195 200 205
Thr Val Ala Pro Thr Glu Cys Ser
210 215
<210> 114
<211> 648
<212> DNA
<213> 人工序列
<220>
<223> IgG 轻链 4
<400> 114
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat agtaataatc agcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgttct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta agtcagccca aggctgcccc ctcggtcacg 360
ctcttcccac cctcctctga ggagcttcaa gccaacaagg ccacactggt gtgtctcata 420
agtgacttct acccgggagc cgtgacagtg gcctggaagg cagatagcag ccccgtcaag 480
gcgggagtgg agaccaccac accctccaaa caaagcaaca acaagtacgc ggccagcagc 540
tacctgagcc tgacgcctga gcagtggaag tcccacagaa gctacagctg ccaggtcacg 600
catgaaggga gcaccgtgga gaagacagtg gcccctacag aatgctct 648
<210> 115
<211> 216
<212> PRT
<213> 人工序列
<220>
<223> IgG 轻链 5
<400> 115
Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln
1 5 10 15
Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ser Asn
20 25 30
Tyr Val Thr Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu
35 40 45
Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser
50 55 60
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln
65 70 75 80
Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ser Asp Glu Leu
85 90 95
Gly Ala Tyr Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln
100 105 110
Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu
115 120 125
Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr
130 135 140
Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys
145 150 155 160
Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr
165 170 175
Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His
180 185 190
Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys
195 200 205
Thr Val Ala Pro Thr Glu Cys Ser
210 215
<210> 116
<211> 648
<212> DNA
<213> 人工序列
<220>
<223> IgG 轻链 5
<400> 116
cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60
tcttgtactg gctcttcatc taatattggc agtaattatg tcacctggta ccagcagctc 120
ccaggaacgg cccccaaact cctcatctat cggaataatc agcggccaag cggggtccct 180
gaccgattct ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag 240
tccgaggatg aggctgatta ttactgttct tcttttagtg atgagttggg ggcttatgtc 300
ttcggcggag gcaccaagct gacggtccta ggtcagccca aggctgcccc ctcggtcacg 360
ctcttcccac cctcctctga ggagcttcaa gccaacaagg ccacactggt gtgtctcata 420
agtgacttct acccgggagc cgtgacagtg gcctggaagg cagatagcag ccccgtcaag 480
gcgggagtgg agaccaccac accctccaaa caaagcaaca acaagtacgc ggccagcagc 540
tacctgagcc tgacgcctga gcagtggaag tcccacagaa gctacagctg ccaggtcacg 600
catgaaggga gcaccgtgga gaagacagtg gcccctacag aatgctct 648
<210> 117
<211> 13
<212> PRT
<213> 人工序列
<220>
<223> N3 Ab 系列的主要结合位点
<400> 117
Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala
1 5 10
<210> 118
<211> 597
<212> PRT
<213> 人工序列
<220>
<223> 赖氨酰-tRNA合成酶的氨基酸序列(KRS, 人)_
Genbank 登录号 . NP_005539.1
<400> 118
Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Ser Glu Pro
1 5 10 15
Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys
20 25 30
Val Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu
35 40 45
Ser Gln Ala Thr Ala Ala Ala Thr Asn His Thr Thr Asp Asn Gly Val
50 55 60
Gly Pro Glu Glu Glu Ser Val Asp Pro Asn Gln Tyr Tyr Lys Ile Arg
65 70 75 80
Ser Gln Ala Ile His Gln Leu Lys Val Asn Gly Glu Asp Pro Tyr Pro
85 90 95
His Lys Phe His Val Asp Ile Ser Leu Thr Asp Phe Ile Gln Lys Tyr
100 105 110
Ser His Leu Gln Pro Gly Asp His Leu Thr Asp Ile Thr Leu Lys Val
115 120 125
Ala Gly Arg Ile His Ala Lys Arg Ala Ser Gly Gly Lys Leu Ile Phe
130 135 140
Tyr Asp Leu Arg Gly Glu Gly Val Lys Leu Gln Val Met Ala Asn Ser
145 150 155 160
Arg Asn Tyr Lys Ser Glu Glu Glu Phe Ile His Ile Asn Asn Lys Leu
165 170 175
Arg Arg Gly Asp Ile Ile Gly Val Gln Gly Asn Pro Gly Lys Thr Lys
180 185 190
Lys Gly Glu Leu Ser Ile Ile Pro Tyr Glu Ile Thr Leu Leu Ser Pro
195 200 205
Cys Leu His Met Leu Pro His Leu His Phe Gly Leu Lys Asp Lys Glu
210 215 220
Thr Arg Tyr Arg Gln Arg Tyr Leu Asp Leu Ile Leu Asn Asp Phe Val
225 230 235 240
Arg Gln Lys Phe Ile Ile Arg Ser Lys Ile Ile Thr Tyr Ile Arg Ser
245 250 255
Phe Leu Asp Glu Leu Gly Phe Leu Glu Ile Glu Thr Pro Met Met Asn
260 265 270
Ile Ile Pro Gly Gly Ala Val Ala Lys Pro Phe Ile Thr Tyr His Asn
275 280 285
Glu Leu Asp Met Asn Leu Tyr Met Arg Ile Ala Pro Glu Leu Tyr His
290 295 300
Lys Met Leu Val Val Gly Gly Ile Asp Arg Val Tyr Glu Ile Gly Arg
305 310 315 320
Gln Phe Arg Asn Glu Gly Ile Asp Leu Thr His Asn Pro Glu Phe Thr
325 330 335
Thr Cys Glu Phe Tyr Met Ala Tyr Ala Asp Tyr His Asp Leu Met Glu
340 345 350
Ile Thr Glu Lys Met Val Ser Gly Met Val Lys His Ile Thr Gly Ser
355 360 365
Tyr Lys Val Thr Tyr His Pro Asp Gly Pro Glu Gly Gln Ala Tyr Asp
370 375 380
Val Asp Phe Thr Pro Pro Phe Arg Arg Ile Asn Met Val Glu Glu Leu
385 390 395 400
Glu Lys Ala Leu Gly Met Lys Leu Pro Glu Thr Asn Leu Phe Glu Thr
405 410 415
Glu Glu Thr Arg Lys Ile Leu Asp Asp Ile Cys Val Ala Lys Ala Val
420 425 430
Glu Cys Pro Pro Pro Arg Thr Thr Ala Arg Leu Leu Asp Lys Leu Val
435 440 445
Gly Glu Phe Leu Glu Val Thr Cys Ile Asn Pro Thr Phe Ile Cys Asp
450 455 460
His Pro Gln Ile Met Ser Pro Leu Ala Lys Trp His Arg Ser Lys Glu
465 470 475 480
Gly Leu Thr Glu Arg Phe Glu Leu Phe Val Met Lys Lys Glu Ile Cys
485 490 495
Asn Ala Tyr Thr Glu Leu Asn Asp Pro Met Arg Gln Arg Gln Leu Phe
500 505 510
Glu Glu Gln Ala Lys Ala Lys Ala Ala Gly Asp Asp Glu Ala Met Phe
515 520 525
Ile Asp Glu Asn Phe Cys Thr Ala Leu Glu Tyr Gly Leu Pro Pro Thr
530 535 540
Ala Gly Trp Gly Met Gly Ile Asp Arg Val Ala Met Phe Leu Thr Asp
545 550 555 560
Ser Asn Asn Ile Lys Glu Val Leu Leu Phe Pro Ala Met Lys Pro Glu
565 570 575
Asp Lys Lys Glu Asn Val Ala Thr Thr Asp Thr Leu Glu Ser Thr Thr
580 585 590
Val Gly Thr Ser Val
595
<210> 119
<211> 1794
<212> DNA
<213> 人工序列
<220>
<223> KRS的DNA序列
<400> 119
atggcggccg tgcaggcggc cgaggtgaaa gtggatggca gcgagccgaa actgagcaag 60
aatgagctga agagacgcct gaaagctgag aagaaagtag cagagaagga ggccaaacag 120
aaagagctca gtgagaaaca gctaagccaa gccactgctg ctgccaccaa ccacaccact 180
gataatggtg tgggtcctga ggaagagagc gtggacccaa atcaatacta caaaatccgc 240
agtcaagcaa ttcatcagct gaaggtcaat ggggaagacc catacccaca caagttccat 300
gtagacatct cactcactga cttcatccaa aaatatagtc acctgcagcc tggggatcac 360
ctgactgaca tcaccttaaa ggtggcaggt aggatccatg ccaaaagagc ttctggggga 420
aagctcatct tctatgatct tcgaggagag ggggtgaagt tgcaagtcat ggccaattcc 480
agaaattata aatcagaaga agaatttatt catattaata acaaactgcg tcggggagac 540
ataattggag ttcaggggaa tcctggtaaa accaagaagg gtgagctgag catcattccg 600
tatgagatca cactgctgtc tccctgtttg catatgttac ctcatcttca ctttgggctc 660
aaagacaagg aaacaaggta tcgccagaga tacttggact tgatcctgaa tgactttgtg 720
aggcagaaat ttatcatccg ctctaagatc atcacatata taagaagttt cttagatgag 780
ctgggattcc tagagattga aactcccatg atgaacatca tcccaggggg agccgtggcc 840
aagcctttca tcacttatca caacgagctg gacatgaact tatatatgag aattgctcca 900
gaactctatc ataagatgct tgtggttggt ggcatcgacc gggtttatga aattggacgc 960
cagttccgga atgaggggat tgatttgacg cacaatcctg agttcaccac ctgtgagttc 1020
tacatggcct atgcagacta tcacgatctc atggaaatca cggagaagat ggtttcaggg 1080
atggtgaagc atattacagg cagttacaag gtcacctacc acccagatgg cccagagggc 1140
caagcctacg atgttgactt caccccaccc ttccggcgaa tcaacatggt agaagagctt 1200
gagaaagccc tggggatgaa gctgccagaa acgaacctct ttgaaactga agaaactcgc 1260
aaaattcttg atgatatctg tgtggcaaaa gctgttgaat gccctccacc tcggaccaca 1320
gccaggctcc ttgacaagct tgttggggag ttcctggaag tgacttgcat caatcctaca 1380
ttcatctgtg atcacccaca gataatgagc cctttggcta aatggcaccg ctctaaagag 1440
ggtctgactg agcgctttga gctgtttgtc atgaagaaag agatatgcaa tgcgtatact 1500
gagctgaatg atcccatgcg gcagcggcag ctttttgaag aacaggccaa ggccaaggct 1560
gcaggtgatg atgaggccat gttcatagat gaaaacttct gtactgccct ggaatatggg 1620
ctgcccccca cagctggctg gggcatgggc attgatcgag tcgccatgtt tctcacggac 1680
tccaacaaca tcaaggaagt acttctgttt cctgccatga aacccgaaga caagaaggag 1740
aatgtagcaa ccactgatac actggaaagc acaacagttg gcacttctgt ctag 1794
<210> 120
<211> 1823
<212> PRT
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元alpha-4的氨基酸序列(人)
<400> 120
Met Ala Leu Ser Ser Ala Trp Arg Ser Val Leu Pro Leu Trp Leu Leu
1 5 10 15
Trp Ser Ala Ala Cys Ser Arg Ala Ala Ser Gly Asp Asp Asn Ala Phe
20 25 30
Pro Phe Asp Ile Glu Gly Ser Ser Ala Val Gly Arg Gln Asp Pro Pro
35 40 45
Glu Thr Ser Glu Pro Arg Val Ala Leu Gly Arg Leu Pro Pro Ala Ala
50 55 60
Glu Lys Cys Asn Ala Gly Phe Phe His Thr Leu Ser Gly Glu Cys Val
65 70 75 80
Pro Cys Asp Cys Asn Gly Asn Ser Asn Glu Cys Leu Asp Gly Ser Gly
85 90 95
Tyr Cys Val His Cys Gln Arg Asn Thr Thr Gly Glu His Cys Glu Lys
100 105 110
Cys Leu Asp Gly Tyr Ile Gly Asp Ser Ile Arg Gly Ala Pro Gln Phe
115 120 125
Cys Gln Pro Cys Pro Cys Pro Leu Pro His Leu Ala Asn Phe Ala Glu
130 135 140
Ser Cys Tyr Arg Lys Asn Gly Ala Val Arg Cys Ile Cys Asn Glu Asn
145 150 155 160
Tyr Ala Gly Pro Asn Cys Glu Arg Cys Ala Pro Gly Tyr Tyr Gly Asn
165 170 175
Pro Leu Leu Ile Gly Ser Thr Cys Lys Lys Cys Asp Cys Ser Gly Asn
180 185 190
Ser Asp Pro Asn Leu Ile Phe Glu Asp Cys Asp Glu Val Thr Gly Gln
195 200 205
Cys Arg Asn Cys Leu Arg Asn Thr Thr Gly Phe Lys Cys Glu Arg Cys
210 215 220
Ala Pro Gly Tyr Tyr Gly Asp Ala Arg Ile Ala Lys Asn Cys Ala Val
225 230 235 240
Cys Asn Cys Gly Gly Gly Pro Cys Asp Ser Val Thr Gly Glu Cys Leu
245 250 255
Glu Glu Gly Phe Glu Pro Pro Thr Gly Met Asp Cys Pro Thr Ile Ser
260 265 270
Cys Asp Lys Cys Val Trp Asp Leu Thr Asp Ala Leu Arg Leu Ala Ala
275 280 285
Leu Ser Ile Glu Glu Gly Lys Ser Gly Val Leu Ser Val Ser Ser Gly
290 295 300
Ala Ala Ala His Arg His Val Asn Glu Ile Asn Ala Thr Ile Tyr Leu
305 310 315 320
Leu Lys Thr Lys Leu Ser Glu Arg Glu Asn Gln Tyr Ala Leu Arg Lys
325 330 335
Ile Gln Ile Asn Asn Ala Glu Asn Thr Met Lys Ser Leu Leu Ser Asp
340 345 350
Val Glu Glu Leu Val Glu Lys Glu Asn Gln Ala Ser Arg Lys Gly Gln
355 360 365
Leu Val Gln Lys Glu Ser Met Asp Thr Ile Asn His Ala Ser Gln Leu
370 375 380
Val Glu Gln Ala His Asp Met Arg Asp Lys Ile Gln Glu Ile Asn Asn
385 390 395 400
Lys Met Leu Tyr Tyr Gly Glu Glu His Glu Leu Ser Pro Lys Glu Ile
405 410 415
Ser Glu Lys Leu Val Leu Ala Gln Lys Met Leu Glu Glu Ile Arg Ser
420 425 430
Arg Gln Pro Phe Phe Thr Gln Arg Glu Leu Val Asp Glu Glu Ala Asp
435 440 445
Glu Ala Tyr Glu Leu Leu Ser Gln Ala Glu Ser Trp Gln Arg Leu His
450 455 460
Asn Glu Thr Arg Thr Leu Phe Pro Val Val Leu Glu Gln Leu Asp Asp
465 470 475 480
Tyr Asn Ala Lys Leu Ser Asp Leu Gln Glu Ala Leu Asp Gln Ala Leu
485 490 495
Asn Tyr Val Arg Asp Ala Glu Asp Met Asn Arg Ala Thr Ala Ala Arg
500 505 510
Gln Arg Asp His Glu Lys Gln Gln Glu Arg Val Arg Glu Gln Met Glu
515 520 525
Val Val Asn Met Ser Leu Ser Thr Ser Ala Asp Ser Leu Thr Thr Pro
530 535 540
Arg Leu Thr Leu Ser Glu Leu Asp Asp Ile Ile Lys Asn Ala Ser Gly
545 550 555 560
Ile Tyr Ala Glu Ile Asp Gly Ala Lys Ser Glu Leu Gln Val Lys Leu
565 570 575
Ser Asn Leu Ser Asn Leu Ser His Asp Leu Val Gln Glu Ala Ile Asp
580 585 590
His Ala Gln Asp Leu Gln Gln Glu Ala Asn Glu Leu Ser Arg Lys Leu
595 600 605
His Ser Ser Asp Met Asn Gly Leu Val Gln Lys Ala Leu Asp Ala Ser
610 615 620
Asn Val Tyr Glu Asn Ile Val Asn Tyr Val Ser Glu Ala Asn Glu Thr
625 630 635 640
Ala Glu Phe Ala Leu Asn Thr Thr Asp Arg Ile Tyr Asp Ala Val Ser
645 650 655
Gly Ile Asp Thr Gln Ile Ile Tyr His Lys Asp Glu Ser Glu Asn Leu
660 665 670
Leu Asn Gln Ala Arg Glu Leu Gln Ala Lys Ala Glu Ser Ser Ser Asp
675 680 685
Glu Ala Val Ala Asp Thr Ser Arg Arg Val Gly Gly Ala Leu Ala Arg
690 695 700
Lys Ser Ala Leu Lys Thr Arg Leu Ser Asp Ala Val Lys Gln Leu Gln
705 710 715 720
Ala Ala Glu Arg Gly Asp Ala Gln Gln Arg Leu Gly Gln Ser Arg Leu
725 730 735
Ile Thr Glu Glu Ala Asn Arg Thr Thr Met Glu Val Gln Gln Ala Thr
740 745 750
Ala Pro Met Ala Asn Asn Leu Thr Asn Trp Ser Gln Asn Leu Gln His
755 760 765
Phe Asp Ser Ser Ala Tyr Asn Thr Ala Val Asn Ser Ala Arg Asp Ala
770 775 780
Val Arg Asn Leu Thr Glu Val Val Pro Gln Leu Leu Asp Gln Leu Arg
785 790 795 800
Thr Val Glu Gln Lys Arg Pro Ala Ser Asn Val Ser Ala Ser Ile Gln
805 810 815
Arg Ile Arg Glu Leu Ile Ala Gln Thr Arg Ser Val Ala Ser Lys Ile
820 825 830
Gln Val Ser Met Met Phe Asp Gly Gln Ser Ala Val Glu Val His Ser
835 840 845
Arg Thr Ser Met Asp Asp Leu Lys Ala Phe Thr Ser Leu Ser Leu Tyr
850 855 860
Met Lys Pro Pro Val Lys Arg Pro Glu Leu Thr Glu Thr Ala Asp Gln
865 870 875 880
Phe Ile Leu Tyr Leu Gly Ser Lys Asn Ala Lys Lys Glu Tyr Met Gly
885 890 895
Leu Ala Ile Lys Asn Asp Asn Leu Val Tyr Val Tyr Asn Leu Gly Thr
900 905 910
Lys Asp Val Glu Ile Pro Leu Asp Ser Lys Pro Val Ser Ser Trp Pro
915 920 925
Ala Tyr Phe Ser Ile Val Lys Ile Glu Arg Val Gly Lys His Gly Lys
930 935 940
Val Phe Leu Thr Val Pro Ser Leu Ser Ser Thr Ala Glu Glu Lys Phe
945 950 955 960
Ile Lys Lys Gly Glu Phe Ser Gly Asp Asp Ser Leu Leu Asp Leu Asp
965 970 975
Pro Glu Asp Thr Val Phe Tyr Val Gly Gly Val Pro Ser Asn Phe Lys
980 985 990
Leu Pro Thr Ser Leu Asn Leu Pro Gly Phe Val Gly Cys Leu Glu Leu
995 1000 1005
Ala Thr Leu Asn Asn Asp Val Ile Ser Leu Tyr Asn Phe Lys His Ile
1010 1015 1020
Tyr Asn Met Asp Pro Ser Thr Ser Val Pro Cys Ala Arg Asp Lys Leu
1025 1030 1035 1040
Ala Phe Thr Gln Ser Arg Ala Ala Ser Tyr Phe Phe Asp Gly Ser Gly
1045 1050 1055
Tyr Ala Val Val Arg Asp Ile Thr Arg Arg Gly Lys Phe Gly Gln Val
1060 1065 1070
Thr Arg Phe Asp Ile Glu Val Arg Thr Pro Ala Asp Asn Gly Leu Ile
1075 1080 1085
Leu Leu Met Val Asn Gly Ser Met Phe Phe Arg Leu Glu Met Arg Asn
1090 1095 1100
Gly Tyr Leu His Val Phe Tyr Asp Phe Gly Phe Ser Gly Gly Pro Val
1105 1110 1115 1120
His Leu Glu Asp Thr Leu Lys Lys Ala Gln Ile Asn Asp Ala Lys Tyr
1125 1130 1135
His Glu Ile Ser Ile Ile Tyr His Asn Asp Lys Lys Met Ile Leu Val
1140 1145 1150
Val Asp Arg Arg His Val Lys Ser Met Asp Asn Glu Lys Met Lys Ile
1155 1160 1165
Pro Phe Thr Asp Ile Tyr Ile Gly Gly Ala Pro Pro Glu Ile Leu Gln
1170 1175 1180
Ser Arg Ala Leu Arg Ala His Leu Pro Leu Asp Ile Asn Phe Arg Gly
1185 1190 1195 1200
Cys Met Lys Gly Phe Gln Phe Gln Lys Lys Asp Phe Asn Leu Leu Glu
1205 1210 1215
Gln Thr Glu Thr Leu Gly Val Gly Tyr Gly Cys Pro Glu Asp Ser Leu
1220 1225 1230
Ile Ser Arg Arg Ala Tyr Phe Asn Gly Gln Ser Phe Ile Ala Ser Ile
1235 1240 1245
Gln Lys Ile Ser Phe Phe Asp Gly Phe Glu Gly Gly Phe Asn Phe Arg
1250 1255 1260
Thr Leu Gln Pro Asn Gly Leu Leu Phe Tyr Tyr Ala Ser Gly Ser Asp
1265 1270 1275 1280
Val Phe Ser Ile Ser Leu Asp Asn Gly Thr Val Ile Met Asp Val Lys
1285 1290 1295
Gly Ile Lys Val Gln Ser Val Asp Lys Gln Tyr Asn Asp Gly Leu Ser
1300 1305 1310
His Phe Val Ile Ser Ser Val Ser Pro Thr Arg Tyr Glu Leu Ile Val
1315 1320 1325
Asp Lys Ser Arg Val Gly Ser Lys Asn Pro Thr Lys Gly Lys Ile Glu
1330 1335 1340
Gln Thr Gln Ala Ser Glu Lys Lys Phe Tyr Phe Gly Gly Ser Pro Ile
1345 1350 1355 1360
Ser Ala Gln Tyr Ala Asn Phe Thr Gly Cys Ile Ser Asn Ala Tyr Phe
1365 1370 1375
Thr Arg Val Asp Arg Asp Val Glu Val Glu Asp Phe Gln Arg Tyr Thr
1380 1385 1390
Glu Lys Val His Thr Ser Leu Tyr Glu Cys Pro Ile Glu Ser Ser Pro
1395 1400 1405
Leu Phe Leu Leu His Lys Lys Gly Lys Asn Leu Ser Lys Pro Lys Ala
1410 1415 1420
Ser Gln Asn Lys Lys Gly Gly Lys Ser Lys Asp Ala Pro Ser Trp Asp
1425 1430 1435 1440
Pro Val Ala Leu Lys Leu Pro Glu Arg Asn Thr Pro Arg Asn Ser His
1445 1450 1455
Cys His Leu Ser Asn Ser Pro Arg Ala Ile Glu His Ala Tyr Gln Tyr
1460 1465 1470
Gly Gly Thr Ala Asn Ser Arg Gln Glu Phe Glu His Leu Lys Gly Asp
1475 1480 1485
Phe Gly Ala Lys Ser Gln Phe Ser Ile Arg Leu Arg Thr Arg Ser Ser
1490 1495 1500
His Gly Met Ile Phe Tyr Val Ser Asp Gln Glu Glu Asn Asp Phe Met
1505 1510 1515 1520
Thr Leu Phe Leu Ala His Gly Arg Leu Val Tyr Met Phe Asn Val Gly
1525 1530 1535
His Lys Lys Leu Lys Ile Arg Ser Gln Glu Lys Tyr Asn Asp Gly Leu
1540 1545 1550
Trp His Asp Val Ile Phe Ile Arg Glu Arg Ser Ser Gly Arg Leu Val
1555 1560 1565
Ile Asp Gly Leu Arg Val Leu Glu Glu Ser Leu Pro Pro Thr Glu Ala
1570 1575 1580
Thr Trp Lys Ile Lys Gly Pro Ile Tyr Leu Gly Gly Val Ala Pro Gly
1585 1590 1595 1600
Lys Ala Val Lys Asn Val Gln Ile Asn Ser Ile Tyr Ser Phe Ser Gly
1605 1610 1615
Cys Leu Ser Asn Leu Gln Leu Asn Gly Ala Ser Ile Thr Ser Ala Ser
1620 1625 1630
Gln Thr Phe Ser Val Thr Pro Cys Phe Glu Gly Pro Met Glu Thr Gly
1635 1640 1645
Thr Tyr Phe Ser Thr Glu Gly Gly Tyr Val Val Leu Asp Glu Ser Phe
1650 1655 1660
Asn Ile Gly Leu Lys Phe Glu Ile Ala Phe Glu Val Arg Pro Arg Ser
1665 1670 1675 1680
Ser Ser Gly Thr Leu Val His Gly His Ser Val Asn Gly Glu Tyr Leu
1685 1690 1695
Asn Val His Met Lys Asn Gly Gln Val Ile Val Lys Val Asn Asn Gly
1700 1705 1710
Ile Arg Asp Phe Ser Thr Ser Val Thr Pro Lys Gln Ser Leu Cys Asp
1715 1720 1725
Gly Arg Trp His Arg Ile Thr Val Ile Arg Asp Ser Asn Val Val Gln
1730 1735 1740
Leu Asp Val Asp Ser Glu Val Asn His Val Val Gly Pro Leu Asn Pro
1745 1750 1755 1760
Lys Pro Ile Asp His Arg Glu Pro Val Phe Val Gly Gly Val Pro Glu
1765 1770 1775
Ser Leu Leu Thr Pro Arg Leu Ala Pro Ser Lys Pro Phe Thr Gly Cys
1780 1785 1790
Ile Arg His Phe Val Ile Asp Gly His Pro Val Ser Phe Ser Lys Ala
1795 1800 1805
Ala Leu Val Ser Gly Ala Val Ser Ile Asn Ser Cys Pro Ala Ala
1810 1815 1820
<210> 121
<211> 5472
<212> DNA
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元alpha-4的DNA序列(人)
<400> 121
atggctttga gctcagcctg gcgctcggtt ctgcctctgt ggctcctctg gagcgctgcc 60
tgctcccgcg ccgcgtccgg ggacgacaac gcttttcctt ttgacattga agggagctca 120
gcggttggca ggcaagaccc gcctgagacg agcgaacccc gcgtggctct gggacgcctg 180
ccgcctgcgg ccgagaaatg caatgctgga ttctttcaca ccctgtcggg agaatgtgtg 240
ccctgcgact gtaatggcaa ttccaacgag tgtttggacg gctcaggata ctgtgtgcac 300
tgccagcgga acacaacagg agagcactgt gaaaagtgtc tggatggtta tatcggagat 360
tccatcaggg gagcacccca attctgccag ccgtgcccct gtcccctgcc ccacttggcc 420
aattttgcag aatcctgcta taggaaaaat ggagctgttc ggtgcatttg taacgaaaat 480
tatgctggac ctaactgtga aagatgtgct cccggttact atggaaaccc cttactcatt 540
ggaagcacct gtaagaaatg tgactgcagt ggaaattcag atcccaacct gatctttgaa 600
gattgtgatg aagtcactgg ccagtgtagg aattgcttac gcaacaccac cggattcaag 660
tgtgaacgtt gcgctcctgg ctactatggg gacgccagga tagccaagaa ctgtgcagtg 720
tgcaactgcg ggggaggccc atgtgacagt gtaaccggag aatgcttgga agaaggtttt 780
gaacccccta caggcatgga ctgcccaacc ataagctgtg ataagtgcgt ctgggacctg 840
actgatgacc tgcggttagc agcgctctcc atcgaggaag gcaaatccgg ggtgctgagc 900
gtatcctctg gggccgccgc tcataggcac gtgaatgaaa tcaacgccac catctacctc 960
ctcaaaacaa aattgtcaga aagagaaaac caatacgccc taagaaagat acaaatcaac 1020
aatgctgaga acacgatgaa aagccttctg tctgacgtag aggaattagt tgaaaaggaa 1080
aatcaagcct ccagaaaagg acaacttgtt cagaaggaaa gcatggacac cattaaccac 1140
gcaagtcagc tggtagagca agcccatgat atgagggata aaatccaaga gatcaacaac 1200
aagatgctct attatgggga agagcatgaa cttagcccca aggaaatctc tgagaagctg 1260
gtgttggccc agaagatgct tgaagagatt agaagccgtc aaccattttt cacccaacgg 1320
gagctcgtgg atgaggaggc agatgaggct tacgaactac tgagccaggc tgagagctgg 1380
cagcggctgc acaatgagac ccgcactctg tttcctgtcg tcctggagca gctggatgac 1440
tacaatgcta agttgtcaga tctccaggaa gcacttgacc aggcccttaa ctatgtcagg 1500
gatgccgaag acatgaacag ggccacagca gccaggcagc gggaccatga gaaacaacag 1560
gaaagagtga gggaacaaat ggaagtggtg aacatgtctc tgagcacatc tgcggactct 1620
ctgacaacac ctcgtctaac tctttcagaa cttgatgata taataaagaa tgcgtcaggg 1680
atttatgcag aaatagatgg agccaaaagt gaactacaag taaaactatc taacctaagt 1740
aacctcagcc atgatttagt ccaagaagct attgaccatg cacaggacct tcaacaagaa 1800
gctaatgaat tgagcaggaa gttgcacagt tcagatatga acgggctggt acagaaggct 1860
ttggatgcat caaatgtcta tgaaaatatt gttaattatg ttagtgaagc caatgaaaca 1920
gcagaatttg ctttgaacac cactgaccga atttatgatg cggtgagtgg gattgatact 1980
caaatcattt accataaaga tgaaagtgag aacctcctca atcaagccag agaactgcaa 2040
gcaaaggcag agtctagcag tgatgaagca gtggctgaca ctagcaggcg tgtgggtgga 2100
gccctagcaa ggaaaagtgc ccttaaaacc agactcagtg atgccgttaa gcaactacaa 2160
gcagcagaga gaggggatgc ccagcagcgc ctggggcagt ctagactgat caccgaggaa 2220
gccaacagga cgacgatgga ggtgcagcag gccactgccc ccatggccaa caatctaacc 2280
aactggtcac agaatcttca acattttgac tcttctgctt acaacactgc agtgaactct 2340
gctagggatg cagtaagaaa tctgaccgag gttgtccctc agctcctgga tcagcttcgt 2400
acggttgagc agaagcgacc tgcaagcaac gtttctgcca gcatccagag gatccgagag 2460
ctcattgctc agaccagaag tgttgccagc aagatccaag tctccatgat gtttgatggc 2520
cagtcagctg tggaagtgca ctcgagaacc agtatggatg acttaaaggc cttcacgtct 2580
ctgagcctgt acatgaaacc ccctgtgaag cggccggaac tgaccgagac tgcagatcag 2640
tttatcctgt acctcggaag caaaaacgcc aaaaaagagt atatgggtct tgcaatcaaa 2700
aatgataatc tggtatacgt ctataatttg ggaactaaag atgtggagat tcccctggac 2760
tccaagcccg tcagttcctg gcctgcttac ttcagcattg tcaagattga aagggtggga 2820
aaacatggaa aggtgttttt aacagtcccg agtctaagta gcacagcaga ggaaaagttc 2880
attaaaaagg gggaattttc gggagatgac tctctgctgg acctggaccc tgaggacaca 2940
gtgttttatg ttggtggagt gccttccaac ttcaagctcc ctaccagctt aaacctgcct 3000
ggctttgttg gctgcctgga actggccact ttgaataatg atgtgatcag cttgtacaac 3060
tttaagcaca tctataatat ggacccctcc acatcagtgc catgtgcccg agataagctg 3120
gccttcactc agagtcgggc tgccagttac ttcttcgatg gctccggtta tgccgtggtg 3180
agagacatca caaggagagg gaaatttggt caggtgactc gctttgacat agaagttcga 3240
acaccagctg acaacggcct tattctcctg atggtcaatg gaagtatgtt tttcagactg 3300
gaaatgcgca atggttacct acatgtgttc tatgattttg gattcagcgg tggccctgtg 3360
catcttgaag atacgttaaa gaaagctcaa attaatgatg caaaatacca tgagatctca 3420
atcatttacc acaatgataa gaaaatgatc ttggtagttg acagaaggca tgtcaagagc 3480
atggataatg aaaagatgaa aatacctttt acagatatat acattggagg agctcctcca 3540
gaaatcttac aatccagggc cctcagagca caccttcccc tagatatcaa cttcagagga 3600
tgcatgaagg gcttccagtt ccaaaagaag gacttcaatt tactggagca gacagaaacc 3660
ctgggagttg gttatggatg cccagaagac tcacttatat ctcgcagagc atatttcaat 3720
ggacagagct tcattgcttc aattcagaaa atatctttct ttgatggctt tgaaggaggt 3780
tttaatttcc gaacattaca accaaatggg ttactattct attatgcttc agggtcagac 3840
gtgttctcca tctcactgga taatggtact gtcatcatgg atgtaaaggg aatcaaagtt 3900
cagtcagtag ataagcagta caatgatggg ctgtcccact tcgtcattag ctctgtctca 3960
cccacaagat atgaactgat agtagataaa agcagagttg ggagtaagaa tcctaccaaa 4020
gggaaaatag aacagacaca agcaagtgaa aagaagtttt acttcggtgg ctcaccaatc 4080
agtgctcagt atgctaattt cactggctgc ataagtaatg cctactttac cagggtggat 4140
agagatgtgg aggttgaaga tttccaacgg tatactgaaa aggtccacac ttctctttat 4200
gagtgtccca ttgagtcttc accattgttt ctcctccata aaaaaggaaa aaatttatcc 4260
aagcctaaag caagtcagaa taaaaaggga gggaaaagta aagatgcacc ttcatgggat 4320
cctgttgctc tgaaactccc agagcggaat actccaagaa actctcattg ccacctttcc 4380
aacagcccta gagcaataga gcacgcctat caatatggag gaacagccaa cagccgccaa 4440
gagtttgaac acttaaaagg agattttggt gccaaatctc agttttccat tcgtctgaga 4500
actcgttcct cccatggcat gatcttctat gtctcagatc aagaagagaa tgacttcatg 4560
actctatttt tggcccatgg ccgcttggtt tacatgttta atgttggtca caaaaaactg 4620
aagattagaa gccaggagaa atacaatgat ggcctgtggc atgatgtgat atttattcga 4680
gaaaggagca gtggccgact ggtaattgat ggtctccgag tcctagaaga aagtcttcct 4740
cctactgaag ctacctggaa aatcaagggt cccatttatt tgggaggtgt ggctcctgga 4800
aaggctgtga aaaatgttca gattaactcc atctacagtt ttagtggctg tctcagcaat 4860
ctccagctca atggggcctc catcacctct gcttctcaga cattcagtgt gaccccttgc 4920
tttgaaggcc ccatggaaac aggaacttac ttttcaacag aaggaggata cgtggttcta 4980
gatgaatctt tcaatattgg attgaagttt gaaattgcat ttgaagtccg tcccagaagc 5040
agttccggaa ccctggtcca cggccacagt gtcaatgggg agtacctaaa tgttcacatg 5100
aaaaatggac aggtcatagt gaaagtcaat aatggcatca gagatttttc cacctcagtt 5160
acacccaagc agagtctctg tgatggcaga tggcacagaa ttacagttat tagagattct 5220
aatgtggttc agttggatgt ggactctgaa gtgaaccatg tggttggacc cctgaatcca 5280
aaaccaattg atcacaggga gcctgtgttt gttggaggtg ttccagaatc tctactgaca 5340
ccacgcttgg cccccagcaa acccttcaca ggctgcatac gccactttgt gattgatgga 5400
cacccagtga gcttcagtaa agcagccctg gtcagcggcg ccgtaagcat caactcctgt 5460
ccagcagcct ga 5472
<210> 122
<211> 1798
<212> PRT
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元beta-2的氨基酸序列(人)
<400> 122
Met Glu Leu Thr Ser Arg Glu Arg Gly Arg Gly Gln Pro Leu Pro Trp
1 5 10 15
Glu Leu Arg Leu Gly Leu Leu Leu Ser Val Leu Ala Ala Thr Leu Ala
20 25 30
Gln Ala Pro Ala Pro Asp Val Pro Gly Cys Ser Arg Gly Ser Cys Tyr
35 40 45
Pro Ala Thr Gly Asp Leu Leu Val Gly Arg Ala Asp Arg Leu Thr Ala
50 55 60
Ser Ser Thr Cys Gly Leu Asn Gly Pro Gln Pro Tyr Cys Ile Val Ser
65 70 75 80
His Leu Gln Asp Glu Lys Lys Cys Phe Leu Cys Asp Ser Arg Arg Pro
85 90 95
Phe Ser Ala Arg Asp Asn Pro His Ser His Arg Ile Gln Asn Val Val
100 105 110
Thr Ser Phe Ala Pro Gln Arg Arg Ala Ala Trp Trp Gln Ser Glu Asn
115 120 125
Gly Ile Pro Ala Val Thr Ile Gln Leu Asp Leu Glu Ala Glu Phe His
130 135 140
Phe Thr His Leu Ile Met Thr Phe Lys Thr Phe Arg Pro Ala Ala Met
145 150 155 160
Leu Val Glu Arg Ser Ala Asp Phe Gly Arg Thr Trp His Val Tyr Arg
165 170 175
Tyr Phe Ser Tyr Asp Cys Gly Ala Asp Phe Pro Gly Val Pro Leu Ala
180 185 190
Pro Pro Arg His Trp Asp Asp Val Val Cys Glu Ser Arg Tyr Ser Glu
195 200 205
Ile Glu Pro Ser Thr Glu Gly Glu Val Ile Tyr Arg Val Leu Asp Pro
210 215 220
Ala Ile Pro Ile Pro Asp Pro Tyr Ser Ser Arg Ile Gln Asn Leu Leu
225 230 235 240
Lys Ile Thr Asn Leu Arg Val Asn Leu Thr Arg Leu His Thr Leu Gly
245 250 255
Asp Asn Leu Leu Asp Pro Arg Arg Glu Ile Arg Glu Lys Tyr Tyr Tyr
260 265 270
Ala Leu Tyr Glu Leu Val Val Arg Gly Asn Cys Phe Cys Tyr Gly His
275 280 285
Ala Ser Glu Cys Ala Pro Ala Pro Gly Ala Pro Ala His Ala Glu Gly
290 295 300
Met Val His Gly Ala Cys Ile Cys Lys His Asn Thr Arg Gly Leu Asn
305 310 315 320
Cys Glu Gln Cys Gln Asp Phe Tyr Arg Asp Leu Pro Trp Arg Pro Ala
325 330 335
Glu Asp Gly His Ser His Ala Cys Arg Lys Cys Glu Cys His Gly His
340 345 350
Thr His Ser Cys His Phe Asp Met Ala Val Tyr Leu Ala Ser Gly Asn
355 360 365
Val Ser Gly Gly Val Cys Asp Gly Cys Gln His Asn Thr Ala Gly Arg
370 375 380
His Cys Glu Leu Cys Arg Pro Phe Phe Tyr Arg Asp Pro Thr Lys Asp
385 390 395 400
Leu Arg Asp Pro Ala Val Cys Arg Ser Cys Asp Cys Asp Pro Met Gly
405 410 415
Ser Gln Asp Gly Gly Arg Cys Asp Ser His Asp Asp Pro Ala Leu Gly
420 425 430
Leu Val Ser Gly Gln Cys Arg Cys Lys Glu His Val Val Gly Thr Arg
435 440 445
Cys Gln Gln Cys Arg Asp Gly Phe Phe Gly Leu Ser Ile Ser Asp Arg
450 455 460
Leu Gly Cys Arg Arg Cys Gln Cys Asn Ala Arg Gly Thr Val Pro Gly
465 470 475 480
Ser Thr Pro Cys Asp Pro Asn Ser Gly Ser Cys Tyr Cys Lys Arg Leu
485 490 495
Val Thr Gly Arg Gly Cys Asp Arg Cys Leu Pro Gly His Trp Gly Leu
500 505 510
Ser His Asp Leu Leu Gly Cys Arg Pro Cys Asp Cys Asp Val Gly Gly
515 520 525
Ala Leu Asp Pro Gln Cys Asp Glu Gly Thr Gly Gln Cys His Cys Arg
530 535 540
Gln His Met Val Gly Arg Arg Cys Glu Gln Val Gln Pro Gly Tyr Phe
545 550 555 560
Arg Pro Phe Leu Asp His Leu Ile Trp Glu Ala Glu Asp Thr Arg Gly
565 570 575
Gln Val Leu Asp Val Val Glu Arg Leu Val Thr Pro Gly Glu Thr Pro
580 585 590
Ser Trp Thr Gly Ser Gly Phe Val Arg Leu Gln Glu Gly Gln Thr Leu
595 600 605
Glu Phe Leu Val Ala Ser Val Pro Lys Ala Met Asp Tyr Asp Leu Leu
610 615 620
Leu Arg Leu Glu Pro Gln Val Pro Glu Gln Trp Ala Glu Leu Glu Leu
625 630 635 640
Ile Val Gln Arg Pro Gly Pro Val Pro Ala His Ser Leu Cys Gly His
645 650 655
Leu Val Pro Lys Asp Asp Arg Ile Gln Gly Thr Leu Gln Pro His Ala
660 665 670
Arg Tyr Leu Ile Phe Pro Asn Pro Val Cys Leu Glu Pro Gly Ile Ser
675 680 685
Tyr Lys Leu His Leu Lys Leu Val Arg Thr Gly Gly Ser Ala Gln Pro
690 695 700
Glu Thr Pro Tyr Ser Gly Pro Gly Leu Leu Ile Asp Ser Leu Val Leu
705 710 715 720
Leu Pro Arg Val Leu Val Leu Glu Met Phe Ser Gly Gly Asp Ala Ala
725 730 735
Ala Leu Glu Arg Gln Ala Thr Phe Glu Arg Tyr Gln Cys His Glu Glu
740 745 750
Gly Leu Val Pro Ser Lys Thr Ser Pro Ser Glu Ala Cys Ala Pro Leu
755 760 765
Leu Ile Ser Leu Ser Thr Leu Ile Tyr Asn Gly Ala Leu Pro Cys Gln
770 775 780
Cys Asn Pro Gln Gly Ser Leu Ser Ser Glu Cys Asn Pro His Gly Gly
785 790 795 800
Gln Cys Leu Cys Lys Pro Gly Val Val Gly Arg Arg Cys Asp Leu Cys
805 810 815
Ala Pro Gly Tyr Tyr Gly Phe Gly Pro Thr Gly Cys Gln Ala Cys Gln
820 825 830
Cys Ser His Glu Gly Ala Leu Ser Ser Leu Cys Glu Lys Thr Ser Gly
835 840 845
Gln Cys Leu Cys Arg Thr Gly Ala Phe Gly Leu Arg Cys Asp Arg Cys
850 855 860
Gln Arg Gly Gln Trp Gly Phe Pro Ser Cys Arg Pro Cys Val Cys Asn
865 870 875 880
Gly His Ala Asp Glu Cys Asn Thr His Thr Gly Ala Cys Leu Gly Cys
885 890 895
Arg Asp His Thr Gly Gly Glu His Cys Glu Arg Cys Ile Ala Gly Phe
900 905 910
His Gly Asp Pro Arg Leu Pro Tyr Gly Gly Gln Cys Arg Pro Cys Pro
915 920 925
Cys Pro Glu Gly Pro Gly Ser Gln Arg His Phe Ala Thr Ser Cys His
930 935 940
Gln Asp Glu Tyr Ser Gln Gln Ile Val Cys His Cys Arg Ala Gly Tyr
945 950 955 960
Thr Gly Leu Arg Cys Glu Ala Cys Ala Pro Gly His Phe Gly Asp Pro
965 970 975
Ser Arg Pro Gly Gly Arg Cys Gln Leu Cys Glu Cys Ser Gly Asn Ile
980 985 990
Asp Pro Met Asp Pro Asp Ala Cys Asp Pro His Thr Gly Gln Cys Leu
995 1000 1005
Arg Cys Leu His His Thr Glu Gly Pro His Cys Ala His Cys Lys Pro
1010 1015 1020
Gly Phe His Gly Gln Ala Ala Arg Gln Ser Cys His Arg Cys Thr Cys
1025 1030 1035 1040
Asn Leu Leu Gly Thr Asn Pro Gln Gln Cys Pro Ser Pro Asp Gln Cys
1045 1050 1055
His Cys Asp Pro Ser Ser Gly Gln Cys Pro Cys Leu Pro Asn Val Gln
1060 1065 1070
Gly Pro Ser Cys Asp Arg Cys Ala Pro Asn Phe Trp Asn Leu Thr Ser
1075 1080 1085
Gly His Gly Cys Gln Pro Cys Ala Cys His Pro Ser Arg Ala Arg Gly
1090 1095 1100
Pro Thr Cys Asn Glu Phe Thr Gly Gln Cys His Cys Arg Ala Gly Phe
1105 1110 1115 1120
Gly Gly Arg Thr Cys Ser Glu Cys Gln Glu Leu His Trp Gly Asp Pro
1125 1130 1135
Gly Leu Gln Cys His Ala Cys Asp Cys Asp Ser Arg Gly Ile Asp Thr
1140 1145 1150
Pro Gln Cys His Arg Phe Thr Gly His Cys Ser Cys Arg Pro Gly Val
1155 1160 1165
Ser Gly Val Arg Cys Asp Gln Cys Ala Arg Gly Phe Ser Gly Ile Phe
1170 1175 1180
Pro Ala Cys His Pro Cys His Ala Cys Phe Gly Asp Trp Asp Arg Val
1185 1190 1195 1200
Val Gln Asp Leu Ala Ala Arg Thr Gln Arg Leu Glu Gln Arg Ala Gln
1205 1210 1215
Glu Leu Gln Gln Thr Gly Val Leu Gly Ala Phe Glu Ser Ser Phe Trp
1220 1225 1230
His Met Gln Glu Lys Leu Gly Ile Val Gln Gly Ile Val Gly Ala Arg
1235 1240 1245
Asn Thr Ser Ala Ala Ser Thr Ala Gln Leu Val Glu Ala Thr Glu Glu
1250 1255 1260
Leu Arg Arg Glu Ile Gly Glu Ala Thr Glu His Leu Thr Gln Leu Glu
1265 1270 1275 1280
Ala Asp Leu Thr Asp Val Gln Asp Glu Asn Phe Asn Ala Asn His Ala
1285 1290 1295
Leu Ser Gly Leu Glu Arg Asp Arg Leu Ala Leu Asn Leu Thr Leu Arg
1300 1305 1310
Gln Leu Asp Gln His Leu Asp Leu Leu Lys His Ser Asn Phe Leu Gly
1315 1320 1325
Ala Tyr Asp Ser Ile Arg His Ala His Ser Gln Ser Ala Glu Ala Glu
1330 1335 1340
Arg Arg Ala Asn Thr Ser Ala Leu Ala Val Pro Ser Pro Val Ser Asn
1345 1350 1355 1360
Ser Ala Ser Ala Arg His Arg Thr Glu Ala Leu Met Asp Ala Gln Lys
1365 1370 1375
Glu Asp Phe Asn Ser Lys His Met Ala Asn Gln Arg Ala Leu Gly Lys
1380 1385 1390
Leu Ser Ala His Thr His Thr Leu Ser Leu Thr Asp Ile Asn Glu Leu
1395 1400 1405
Val Cys Gly Ala Pro Gly Asp Ala Pro Cys Ala Thr Ser Pro Cys Gly
1410 1415 1420
Gly Ala Gly Cys Arg Asp Glu Asp Gly Gln Pro Arg Cys Gly Gly Leu
1425 1430 1435 1440
Ser Cys Asn Gly Ala Ala Ala Thr Ala Asp Leu Ala Leu Gly Arg Ala
1445 1450 1455
Arg His Thr Gln Ala Glu Leu Gln Arg Ala Leu Ala Glu Gly Gly Ser
1460 1465 1470
Ile Leu Ser Arg Val Ala Glu Thr Arg Arg Gln Ala Ser Glu Ala Gln
1475 1480 1485
Gln Arg Ala Gln Ala Ala Leu Asp Lys Ala Asn Ala Ser Arg Gly Gln
1490 1495 1500
Val Glu Gln Ala Asn Gln Glu Leu Gln Glu Leu Ile Gln Ser Val Lys
1505 1510 1515 1520
Asp Phe Leu Asn Gln Glu Gly Ala Asp Pro Asp Ser Ile Glu Met Val
1525 1530 1535
Ala Thr Arg Val Leu Glu Leu Ser Ile Pro Ala Ser Ala Glu Gln Ile
1540 1545 1550
Gln His Leu Ala Gly Ala Ile Ala Glu Arg Val Arg Ser Leu Ala Asp
1555 1560 1565
Val Asp Ala Ile Leu Ala Arg Thr Val Gly Asp Val Arg Arg Ala Glu
1570 1575 1580
Gln Leu Leu Gln Asp Ala Arg Arg Ala Arg Ser Trp Ala Glu Asp Glu
1585 1590 1595 1600
Lys Gln Lys Ala Glu Thr Val Gln Ala Ala Leu Glu Glu Ala Gln Arg
1605 1610 1615
Ala Gln Gly Ile Ala Gln Gly Ala Ile Arg Gly Ala Val Ala Asp Thr
1620 1625 1630
Arg Asp Thr Glu Gln Thr Leu Tyr Gln Val Gln Glu Arg Met Ala Gly
1635 1640 1645
Ala Glu Arg Ala Leu Ser Ser Ala Gly Glu Arg Ala Arg Gln Leu Asp
1650 1655 1660
Ala Leu Leu Glu Ala Leu Lys Leu Lys Arg Ala Gly Asn Ser Leu Ala
1665 1670 1675 1680
Ala Ser Thr Ala Glu Glu Thr Ala Gly Ser Ala Gln Gly Arg Ala Gln
1685 1690 1695
Glu Ala Glu Gln Leu Leu Arg Gly Pro Leu Gly Asp Gln Tyr Gln Thr
1700 1705 1710
Val Lys Ala Leu Ala Glu Arg Lys Ala Gln Gly Val Leu Ala Ala Gln
1715 1720 1725
Ala Arg Ala Glu Gln Leu Arg Asp Glu Ala Arg Asp Leu Leu Gln Ala
1730 1735 1740
Ala Gln Asp Lys Leu Gln Arg Leu Gln Glu Leu Glu Gly Thr Tyr Glu
1745 1750 1755 1760
Glu Asn Glu Arg Ala Leu Glu Ser Lys Ala Ala Gln Leu Asp Gly Leu
1765 1770 1775
Glu Ala Arg Met Arg Ser Val Leu Gln Ala Ile Asn Leu Gln Val Gln
1780 1785 1790
Ile Tyr Asn Thr Cys Gln
1795
<210> 123
<211> 5397
<212> DNA
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元beta-2的DNA序列(人)
<400> 123
atggagctga cctcaaggga aagagggagg ggacagcctc tgccctggga acttcgactg 60
ggcctactgc taagcgtgct ggctgccaca ctggcacagg cccctgcccc ggatgtgcct 120
ggctgttcca ggggaagctg ctaccccgcc acgggcgacc tgctggtggg ccgagctgac 180
agactgactg cctcatccac ttgtggcctg aatggccccc agccctactg catcgtcagt 240
cacctgcagg acgaaaagaa gtgcttcctt tgtgactccc ggcgcccctt ctctgctaga 300
gacaacccac acagccatcg catccagaat gtagtcacca gctttgcacc acagcggcgg 360
gcagcctggt ggcagtcaga gaatggtatc cctgcggtca ccatccagct ggacctggag 420
gctgagtttc atttcacaca cctcattatg accttcaaga catttcgccc tgctgccatg 480
ctggtggaac gctcagcaga ctttggccgc acctggcatg tgtaccgata tttctcctat 540
gactgtgggg ctgacttccc aggagtccca ctagcacccc cacggcactg ggatgatgta 600
gtctgtgagt cccgctactc agagattgag ccatccactg aaggcgaggt catctatcgt 660
gtgctggacc ctgccatccc tatcccagac ccctacagct cacggattca gaacctgttg 720
aagatcacca acctacgggt gaacctgact cgtctacaca cgttgggaga caacctactc 780
gacccacgga gggagatccg agagaagtac tactatgccc tctatgagct ggttgtacgt 840
ggcaactgct tctgctacgg acacgcctca gagtgtgcac ccgccccagg ggcaccagcc 900
catgctgagg gcatggtgca cggagcttgc atctgcaaac acaacacacg tggcctcaac 960
tgcgagcagt gtcaggattt ctatcgtgac ctgccctggc gtccggctga ggacggccat 1020
agtcatgcct gtaggaagtg tgagtgccat gggcacaccc acagctgcca cttcgacatg 1080
gccgtatacc tggcatctgg caatgtgagt ggaggtgtgt gtgatggatg tcagcataac 1140
acagctgggc gccactgtga gctctgtcgg cccttcttct accgtgaccc aaccaaggac 1200
ctgcgggatc cggctgtgtg ccgctcctgt gattgtgacc ccatgggttc tcaagacggt 1260
ggtcgctgtg attcccatga tgaccctgca ctgggactgg tctccggcca gtgtcgctgc 1320
aaagaacatg tggtgggcac tcgctgccag caatgccgtg atggcttctt tgggctcagc 1380
atcagtgacc gtctgggctg ccggcgatgt caatgtaatg cacggggcac agtgcctggg 1440
agcactcctt gtgaccccaa cagtggatcc tgttactgca aacgtctagt gactggacgt 1500
ggatgtgacc gctgcctgcc tggccactgg ggcctgagcc acgacctgct cggctgccgc 1560
ccctgtgact gcgacgtggg tggtgctttg gatccccagt gtgatgaggg cacaggtcaa 1620
tgccactgcc gccagcacat ggttgggcga cgctgtgagc aggtgcaacc tggctacttc 1680
cggcccttcc tggaccacct aatttgggag gctgaggaca cccgagggca ggtgctcgat 1740
gtggtggagc gcctggtgac ccccggggaa actccatcct ggactggctc aggcttcgtg 1800
cggctacagg aaggtcagac cctggagttc ctggtggcct ctgtgccgaa ggctatggac 1860
tatgacctgc tgctgcgctt agagccccag gtccctgagc aatgggcaga gttggaactg 1920
attgtgcagc gtccagggcc tgtgcctgcc cacagcctgt gtgggcattt ggtgcccaag 1980
gatgatcgca tccaagggac tctgcaacca catgccaggt acttgatatt tcctaatcct 2040
gtctgccttg agcctggtat ctcctacaag ctgcatctga agctggtacg gacaggggga 2100
agtgcccagc ctgagactcc ctactctgga cctggcctgc tcattgactc gctggtgctg 2160
ctgccccgtg tcctggtgct agagatgttt agtgggggtg atgctgctgc cctggagcgc 2220
caggccacct ttgaacgcta ccaatgccat gaggagggtc tggtgcccag caagacttct 2280
ccctctgagg cctgcgcacc cctcctcatc agcctgtcca ccctcatcta caatggtgcc 2340
ctgccatgtc agtgcaaccc tcaaggttca ctgagttctg agtgcaaccc tcatggtggt 2400
cagtgcctgt gcaagcctgg agtggttggg cgccgctgtg acctctgtgc ccctggctac 2460
tatggctttg gccccacagg ctgtcaagcc tgccagtgca gccacgaggg ggcactcagc 2520
agtctctgtg aaaagaccag tgggcaatgt ctctgtcgaa ctggtgcctt tgggcttcgc 2580
tgtgaccgct gccagcgtgg ccagtgggga ttccctagct gccggccatg tgtctgcaat 2640
gggcatgcag atgagtgcaa cacccacaca ggcgcttgcc tgggctgccg tgatcacaca 2700
gggggtgagc actgtgaaag gtgcattgct ggtttccacg gggacccacg gctgccatat 2760
gggggccagt gccggccctg tccctgtcct gaaggccctg ggagccaacg gcactttgct 2820
acttcttgcc accaggatga atattcccag cagattgtgt gccactgccg ggcaggctat 2880
acggggctgc gatgtgaagc ttgtgcccct gggcactttg gggacccatc aaggccaggt 2940
ggccggtgcc aactgtgtga gtgcagtggg aacattgacc caatggatcc tgatgcctgt 3000
gacccccaca cggggcaatg cctgcgctgt ttacaccaca cagagggtcc acactgtgcc 3060
cactgcaagc ctggcttcca tgggcaggct gcccgacaga gctgtcaccg ctgcacatgc 3120
aacctgctgg gcacaaatcc gcagcagtgc ccatctcctg accagtgcca ctgtgatcca 3180
agcagtgggc agtgcccatg cctccccaat gtccagggcc ctagctgtga ccgctgtgcc 3240
cccaacttct ggaacctcac cagtggccat ggttgccagc cttgtgcctg ccacccaagc 3300
cgggccagag gccccacctg caacgagttc acagggcagt gccactgccg tgccggcttt 3360
ggagggcgga cttgttctga gtgccaagag ctccactggg gagaccctgg gttgcagtgc 3420
catgcctgtg attgtgactc tcgtggaata gatacacctc agtgtcaccg cttcacaggt 3480
cactgcagct gccgcccagg ggtgtctggt gtgcgctgtg accagtgtgc ccgtggcttc 3540
tcaggaatct ttcctgcctg ccatccctgc catgcatgct tcggggattg ggaccgagtg 3600
gtgcaggact tggcagcccg tacacagcgc ctagagcagc gggcgcagga gttgcaacag 3660
acgggtgtgc tgggtgcctt tgagagcagc ttctggcaca tgcaggagaa gctgggcatt 3720
gtgcagggca tcgtaggtgc ccgcaacacc tcagccgcct ccactgcaca gcttgtggag 3780
gccacagagg agctgcggcg tgaaattggg gaggccactg agcacctgac tcagctcgag 3840
gcagacctga cagatgtgca agatgagaac ttcaatgcca accatgcact aagtggtctg 3900
gagcgagata ggcttgcact taatctcaca ctgcggcagc tcgaccagca tcttgacttg 3960
ctcaaacatt caaacttcct gggtgcctat gacagcatcc ggcatgccca tagccagtct 4020
gcagaggcag aacgtcgtgc caatacctca gccctggcag tacctagccc tgtgagcaac 4080
tcggcaagtg ctcggcatcg gacagaggca ctgatggatg ctcagaagga ggacttcaac 4140
agcaaacaca tggccaacca gcgggcactt ggcaagctct ctgcccatac ccacaccctg 4200
agcctgacag acataaatga gctggtgtgt ggggcaccag gggatgcacc ctgtgctaca 4260
agcccttgtg ggggtgccgg ctgtcgagat gaggatgggc agccgcgctg tgggggcctc 4320
agctgcaatg gggcagcggc tacagcagac ctagcactgg gccgggcccg gcacacacag 4380
gcagagctgc agcgggcact ggcagaaggt ggtagcatcc tcagcagagt ggctgagact 4440
cgtcggcagg caagcgaggc acagcagcgg gcccaggcag ccctggacaa ggctaatgct 4500
tccaggggac aggtggaaca ggccaaccag gaacttcaag aacttatcca gagtgtgaag 4560
gacttcctca accaggaggg ggctgatcct gatagcattg aaatggtggc cacacgggtg 4620
ctagagctct ccatcccagc ttcagctgag cagatccagc acctggcggg tgcgattgca 4680
gagcgagtcc ggagcctggc agatgtggat gcgatcctgg cacgtactgt aggagatgtg 4740
cgtcgtgccg agcagctact gcaggatgca cggcgggcaa ggagctgggc tgaggatgag 4800
aaacagaagg cagagacagt acaggcagca ctggaggagg cccagcgggc acagggtatt 4860
gcccagggtg ccatccgggg ggcagtggct gacacacggg acacagagca gaccctgtac 4920
caggtacagg agaggatggc aggtgcagag cgggcactga gctctgcagg tgaaagggct 4980
cggcagttgg atgctctcct ggaggctctg aaattgaaac gggcaggaaa tagtctggca 5040
gcctctacag cagaagaaac ggcaggcagt gcccagggtc gtgcccagga ggctgagcag 5100
ctgctacgcg gtcctctggg tgatcagtac cagacggtga aggccctagc tgagcgcaag 5160
gcccaaggtg tgctggctgc acaggcaagg gcagaacaac tgcgggatga ggctcgggac 5220
ctgttgcaag ccgctcagga caagctgcag cggctacagg aattggaagg cacctatgag 5280
gaaaatgagc gggcactgga gagtaaggca gcccagttgg acgggttgga ggccaggatg 5340
cgcagcgtgc ttcaagccat caacttgcag gtgcagatct acaacacctg ccagtga 5397
<210> 124
<211> 1609
<212> PRT
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元gamma-1的氨基酸序列(人)
<400> 124
Met Arg Gly Ser His Arg Ala Ala Pro Ala Leu Arg Pro Arg Gly Arg
1 5 10 15
Leu Trp Pro Val Leu Ala Val Leu Ala Ala Ala Ala Ala Ala Gly Cys
20 25 30
Ala Gln Ala Ala Met Asp Glu Cys Thr Asp Glu Gly Gly Arg Pro Gln
35 40 45
Arg Cys Met Pro Glu Phe Val Asn Ala Ala Phe Asn Val Thr Val Val
50 55 60
Ala Thr Asn Thr Cys Gly Thr Pro Pro Glu Glu Tyr Cys Val Gln Thr
65 70 75 80
Gly Val Thr Gly Val Thr Lys Ser Cys His Leu Cys Asp Ala Gly Gln
85 90 95
Pro His Leu Gln His Gly Ala Ala Phe Leu Thr Asp Tyr Asn Asn Gln
100 105 110
Ala Asp Thr Thr Trp Trp Gln Ser Gln Thr Met Leu Ala Gly Val Gln
115 120 125
Tyr Pro Ser Ser Ile Asn Leu Thr Leu His Leu Gly Lys Ala Phe Asp
130 135 140
Ile Thr Tyr Val Arg Leu Lys Phe His Thr Ser Arg Pro Glu Ser Phe
145 150 155 160
Ala Ile Tyr Lys Arg Thr Arg Glu Asp Gly Pro Trp Ile Pro Tyr Gln
165 170 175
Tyr Tyr Ser Gly Ser Cys Glu Asn Thr Tyr Ser Lys Ala Asn Arg Gly
180 185 190
Phe Ile Arg Thr Gly Gly Asp Glu Gln Gln Ala Leu Cys Thr Asp Glu
195 200 205
Phe Ser Asp Ile Ser Pro Leu Thr Gly Gly Asn Val Ala Phe Ser Thr
210 215 220
Leu Glu Gly Arg Pro Ser Ala Tyr Asn Phe Asp Asn Ser Pro Val Leu
225 230 235 240
Gln Glu Trp Val Thr Ala Thr Asp Ile Arg Val Thr Leu Asn Arg Leu
245 250 255
Asn Thr Phe Gly Asp Glu Val Phe Asn Asp Pro Lys Val Leu Lys Ser
260 265 270
Tyr Tyr Tyr Ala Ile Ser Asp Phe Ala Val Gly Gly Arg Cys Lys Cys
275 280 285
Asn Gly His Ala Ser Glu Cys Met Lys Asn Glu Phe Asp Lys Leu Val
290 295 300
Cys Asn Cys Lys His Asn Thr Tyr Gly Val Asp Cys Glu Lys Cys Leu
305 310 315 320
Pro Phe Phe Asn Asp Arg Pro Trp Arg Arg Ala Thr Ala Glu Ser Ala
325 330 335
Ser Glu Cys Leu Pro Cys Asp Cys Asn Gly Arg Ser Gln Glu Cys Tyr
340 345 350
Phe Asp Pro Glu Leu Tyr Arg Ser Thr Gly His Gly Gly His Cys Thr
355 360 365
Asn Cys Gln Asp Asn Thr Asp Gly Ala His Cys Glu Arg Cys Arg Glu
370 375 380
Asn Phe Phe Arg Leu Gly Asn Asn Glu Ala Cys Ser Ser Cys His Cys
385 390 395 400
Ser Pro Val Gly Ser Leu Ser Thr Gln Cys Asp Ser Tyr Gly Arg Cys
405 410 415
Ser Cys Lys Pro Gly Val Met Gly Asp Lys Cys Asp Arg Cys Gln Pro
420 425 430
Gly Phe His Ser Leu Thr Glu Ala Gly Cys Arg Pro Cys Ser Cys Asp
435 440 445
Pro Ser Gly Ser Ile Asp Glu Cys Asn Ile Glu Thr Gly Arg Cys Val
450 455 460
Cys Lys Asp Asn Val Glu Gly Phe Asn Cys Glu Arg Cys Lys Pro Gly
465 470 475 480
Phe Phe Asn Leu Glu Ser Ser Asn Pro Arg Gly Cys Thr Pro Cys Phe
485 490 495
Cys Phe Gly His Ser Ser Val Cys Thr Asn Ala Val Gly Tyr Ser Val
500 505 510
Tyr Ser Ile Ser Ser Thr Phe Gln Ile Asp Glu Asp Gly Trp Arg Ala
515 520 525
Glu Gln Arg Asp Gly Ser Glu Ala Ser Leu Glu Trp Ser Ser Glu Arg
530 535 540
Gln Asp Ile Ala Val Ile Ser Asp Ser Tyr Phe Pro Arg Tyr Phe Ile
545 550 555 560
Ala Pro Ala Lys Phe Leu Gly Lys Gln Val Leu Ser Tyr Gly Gln Asn
565 570 575
Leu Ser Phe Ser Phe Arg Val Asp Arg Arg Asp Thr Arg Leu Ser Ala
580 585 590
Glu Asp Leu Val Leu Glu Gly Ala Gly Leu Arg Val Ser Val Pro Leu
595 600 605
Ile Ala Gln Gly Asn Ser Tyr Pro Ser Glu Thr Thr Val Lys Tyr Val
610 615 620
Phe Arg Leu His Glu Ala Thr Asp Tyr Pro Trp Arg Pro Ala Leu Thr
625 630 635 640
Pro Phe Glu Phe Gln Lys Leu Leu Asn Asn Leu Thr Ser Ile Lys Ile
645 650 655
Arg Gly Thr Tyr Ser Glu Arg Ser Ala Gly Tyr Leu Asp Asp Val Thr
660 665 670
Leu Ala Ser Ala Arg Pro Gly Pro Gly Val Pro Ala Thr Trp Val Glu
675 680 685
Ser Cys Thr Cys Pro Val Gly Tyr Gly Gly Gln Phe Cys Glu Met Cys
690 695 700
Leu Ser Gly Tyr Arg Arg Glu Thr Pro Asn Leu Gly Pro Tyr Ser Pro
705 710 715 720
Cys Val Leu Cys Ala Cys Asn Gly His Ser Glu Thr Cys Asp Pro Glu
725 730 735
Thr Gly Val Cys Asn Cys Arg Asp Asn Thr Ala Gly Pro His Cys Glu
740 745 750
Lys Cys Ser Asp Gly Tyr Tyr Gly Asp Ser Thr Ala Gly Thr Ser Ser
755 760 765
Asp Cys Gln Pro Cys Pro Cys Pro Gly Gly Ser Ser Cys Ala Val Val
770 775 780
Pro Lys Thr Lys Glu Val Val Cys Thr Asn Cys Pro Thr Gly Thr Thr
785 790 795 800
Gly Lys Arg Cys Glu Leu Cys Asp Asp Gly Tyr Phe Gly Asp Pro Leu
805 810 815
Gly Arg Asn Gly Pro Val Arg Leu Cys Arg Leu Cys Gln Cys Ser Asp
820 825 830
Asn Ile Asp Pro Asn Ala Val Gly Asn Cys Asn Arg Leu Thr Gly Glu
835 840 845
Cys Leu Lys Cys Ile Tyr Asn Thr Ala Gly Phe Tyr Cys Asp Arg Cys
850 855 860
Lys Asp Gly Phe Phe Gly Asn Pro Leu Ala Pro Asn Pro Ala Asp Lys
865 870 875 880
Cys Lys Ala Cys Asn Cys Asn Leu Tyr Gly Thr Met Lys Gln Gln Ser
885 890 895
Ser Cys Asn Pro Val Thr Gly Gln Cys Glu Cys Leu Pro His Val Thr
900 905 910
Gly Gln Asp Cys Gly Ala Cys Asp Pro Gly Phe Tyr Asn Leu Gln Ser
915 920 925
Gly Gln Gly Cys Glu Arg Cys Asp Cys His Ala Leu Gly Ser Thr Asn
930 935 940
Gly Gln Cys Asp Ile Arg Thr Gly Gln Cys Glu Cys Gln Pro Gly Ile
945 950 955 960
Thr Gly Gln His Cys Glu Arg Cys Glu Val Asn His Phe Gly Phe Gly
965 970 975
Pro Glu Gly Cys Lys Pro Cys Asp Cys His Pro Glu Gly Ser Leu Ser
980 985 990
Leu Gln Cys Lys Asp Asp Gly Arg Cys Glu Cys Arg Glu Gly Phe Val
995 1000 1005
Gly Asn Arg Cys Asp Gln Cys Glu Glu Asn Tyr Phe Tyr Asn Arg Ser
1010 1015 1020
Trp Pro Gly Cys Gln Glu Cys Pro Ala Cys Tyr Arg Leu Val Lys Asp
1025 1030 1035 1040
Lys Val Ala Asp His Arg Val Lys Leu Gln Glu Leu Glu Ser Leu Ile
1045 1050 1055
Ala Asn Leu Gly Thr Gly Asp Glu Met Val Thr Asp Gln Ala Phe Glu
1060 1065 1070
Asp Arg Leu Lys Glu Ala Glu Arg Glu Val Met Asp Leu Leu Arg Glu
1075 1080 1085
Ala Gln Asp Val Lys Asp Val Asp Gln Asn Leu Met Asp Arg Leu Gln
1090 1095 1100
Arg Val Asn Asn Thr Leu Ser Ser Gln Ile Ser Arg Leu Gln Asn Ile
1105 1110 1115 1120
Arg Asn Thr Ile Glu Glu Thr Gly Asn Leu Ala Glu Gln Ala Arg Ala
1125 1130 1135
His Val Glu Asn Thr Glu Arg Leu Ile Glu Ile Ala Ser Arg Glu Leu
1140 1145 1150
Glu Lys Ala Lys Val Ala Ala Ala Asn Val Ser Val Thr Gln Pro Glu
1155 1160 1165
Ser Thr Gly Asp Pro Asn Asn Met Thr Leu Leu Ala Glu Glu Ala Arg
1170 1175 1180
Lys Leu Ala Glu Arg His Lys Gln Glu Ala Asp Asp Ile Val Arg Val
1185 1190 1195 1200
Ala Lys Thr Ala Asn Asp Thr Ser Thr Glu Ala Tyr Asn Leu Leu Leu
1205 1210 1215
Arg Thr Leu Ala Gly Glu Asn Gln Thr Ala Phe Glu Ile Glu Glu Leu
1220 1225 1230
Asn Arg Lys Tyr Glu Gln Ala Lys Asn Ile Ser Gln Asp Leu Glu Lys
1235 1240 1245
Gln Ala Ala Arg Val His Glu Glu Ala Lys Arg Ala Gly Asp Lys Ala
1250 1255 1260
Val Glu Ile Tyr Ala Ser Val Ala Gln Leu Ser Pro Leu Asp Ser Glu
1265 1270 1275 1280
Thr Leu Glu Asn Glu Ala Asn Asn Ile Lys Met Glu Ala Glu Asn Leu
1285 1290 1295
Glu Gln Leu Ile Asp Gln Lys Leu Lys Asp Tyr Glu Asp Leu Arg Glu
1300 1305 1310
Asp Met Arg Gly Lys Glu Leu Glu Val Lys Asn Leu Leu Glu Lys Gly
1315 1320 1325
Lys Thr Glu Gln Gln Thr Ala Asp Gln Leu Leu Ala Arg Ala Asp Ala
1330 1335 1340
Ala Lys Ala Leu Ala Glu Glu Ala Ala Lys Lys Gly Arg Asp Thr Leu
1345 1350 1355 1360
Gln Glu Ala Asn Asp Ile Leu Asn Asn Leu Lys Asp Phe Asp Arg Arg
1365 1370 1375
Val Asn Asp Asn Lys Thr Ala Ala Glu Glu Ala Leu Arg Lys Ile Pro
1380 1385 1390
Ala Ile Asn Gln Thr Ile Thr Glu Ala Asn Glu Lys Thr Arg Glu Ala
1395 1400 1405
Gln Gln Ala Leu Gly Ser Ala Ala Ala Asp Ala Thr Glu Ala Lys Asn
1410 1415 1420
Lys Ala His Glu Ala Glu Arg Ile Ala Ser Ala Val Gln Lys Asn Ala
1425 1430 1435 1440
Thr Ser Thr Lys Ala Glu Ala Glu Arg Thr Phe Ala Glu Val Thr Asp
1445 1450 1455
Leu Asp Asn Glu Val Asn Asn Met Leu Lys Gln Leu Gln Glu Ala Glu
1460 1465 1470
Lys Glu Leu Lys Arg Lys Gln Asp Asp Ala Asp Gln Asp Met Met Met
1475 1480 1485
Ala Gly Met Ala Ser Gln Ala Ala Gln Glu Ala Glu Ile Asn Ala Arg
1490 1495 1500
Lys Ala Lys Asn Ser Val Thr Ser Leu Leu Ser Ile Ile Asn Asp Leu
1505 1510 1515 1520
Leu Glu Gln Leu Gly Gln Leu Asp Thr Val Asp Leu Asn Lys Leu Asn
1525 1530 1535
Glu Ile Glu Gly Thr Leu Asn Lys Ala Lys Asp Glu Met Lys Val Ser
1540 1545 1550
Asp Leu Asp Arg Lys Val Ser Asp Leu Glu Asn Glu Ala Lys Lys Gln
1555 1560 1565
Glu Ala Ala Ile Met Asp Tyr Asn Arg Asp Ile Glu Glu Ile Met Lys
1570 1575 1580
Asp Ile Arg Asn Leu Glu Asp Ile Arg Lys Thr Leu Pro Ser Gly Cys
1585 1590 1595 1600
Phe Asn Thr Pro Ser Ile Glu Lys Pro
1605
<210> 125
<211> 4830
<212> DNA
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元gamma-1的DNA序列(人)
<400> 125
atgagaggga gccatcgggc cgcgccggcc ctgcggcccc gggggcggct ctggcccgtg 60
ctggccgtgc tggcggcggc cgccgcggcg ggctgtgccc aggcagccat ggacgagtgc 120
acggacgagg gcgggcggcc gcagcgctgc atgcccgagt tcgtcaacgc cgccttcaac 180
gtgactgtgg tggccaccaa cacgtgtggg actccgcccg aggaatactg tgtgcagacc 240
ggggtgaccg gggtcaccaa gtcctgtcac ctgtgcgacg ccgggcagcc ccacctgcag 300
cacggggcag ccttcctgac cgactacaac aaccaggccg acaccacctg gtggcaaagc 360
cagaccatgc tggccggggt gcagtacccc agctccatca acctcacgct gcacctggga 420
aaagcttttg acatcaccta tgtgcgtctc aagttccaca ccagccgccc ggagagcttt 480
gccatttaca agcgcacacg ggaagacggg ccctggattc cttaccagta ctacagtggt 540
tcctgtgaga acacctactc caaggcaaac cgcggcttca tcaggacagg aggggacgag 600
cagcaggcct tgtgtactga tgaattcagt gacatttctc ccctcactgg gggcaacgtg 660
gccttttcta ccctggaagg aaggcccagc gcctataact ttgacaatag ccctgtgctg 720
caggaatggg taactgccac tgacatcaga gtaactctta atcgcctgaa cacttttgga 780
gatgaagtgt ttaacgatcc caaagttctc aagtcctatt attatgccat ctctgatttt 840
gctgtaggtg gcagatgtaa atgtaatgga cacgcaagcg agtgtatgaa gaacgaattt 900
gataagctgg tgtgtaattg caaacataac acatatggag tagactgtga aaagtgtctt 960
cctttcttca atgaccggcc gtggaggagg gcaactgcgg aaagtgccag tgaatgcctg 1020
ccctgtgatt gcaatggtcg atcccaggaa tgctacttcg accctgaact ctatcgttcc 1080
actggccatg ggggccactg taccaactgc caggataaca cagatggcgc ccactgtgag 1140
aggtgccgag agaacttctt ccgccttggc aacaatgaag cctgctcttc atgccactgt 1200
agtcctgtgg gctctctaag cacacagtgt gatagttacg gcagatgcag ctgtaagcca 1260
ggagtgatgg gggacaaatg tgaccgttgc cagcctggat tccattctct cactgaagca 1320
ggatgcaggc catgctcttg tgatccctct ggcagcatag atgaatgtaa tattgaaaca 1380
ggaagatgtg tttgcaaaga caatgtcgaa ggcttcaatt gtgaaagatg caaacctgga 1440
ttttttaatc tggaatcatc taatcctcgg ggttgcacac cctgcttctg ctttgggcat 1500
tcttctgtct gtacaaacgc tgttggctac agtgtttatt ctatctcctc tacctttcag 1560
attgatgagg atgggtggcg tgcggaacag agagatggct ctgaagcatc tctcgagtgg 1620
tcctctgaga ggcaagatat cgccgtgatc tcagacagct actttcctcg gtacttcatt 1680
gctcctgcaa agttcttggg caagcaggtg ttgagttatg gtcagaacct ctccttctcc 1740
tttcgagtgg acaggcgaga tactcgcctc tctgcagaag accttgtgct tgagggagct 1800
ggcttaagag tatctgtacc cttgatcgct cagggcaatt cctatccaag tgagaccact 1860
gtgaagtatg tcttcaggct ccatgaagca acagattacc cttggaggcc tgctcttacc 1920
ccttttgaat ttcagaagct cctaaacaac ttgacctcta tcaagatacg tgggacatac 1980
agtgagagaa gtgctggata tttggatgat gtcaccctgg caagtgctcg tcctgggcct 2040
ggagtccctg caacttgggt ggagtcctgc acctgtcctg tgggatatgg agggcagttt 2100
tgtgagatgt gcctctcagg ttacagaaga gaaactccta atcttggacc atacagtcca 2160
tgtgtgcttt gcgcctgcaa tggacacagc gagacctgtg atcctgagac aggtgtttgt 2220
aactgcagag acaatacggc tggcccgcac tgtgagaagt gcagtgatgg gtactatgga 2280
gattcaactg caggcacctc ctccgattgc caaccctgtc cgtgtcctgg aggttcaagt 2340
tgtgctgttg ttcccaagac aaaggaggtg gtgtgcacca actgtcctac tggcaccact 2400
ggtaagagat gtgagctctg tgatgatggc tactttggag accccctggg tagaaacggc 2460
cctgtgagac tttgccgcct gtgccagtgc agtgacaaca tcgatcccaa tgcagttgga 2520
aattgcaatc gcttgacggg agaatgcctg aagtgcatct ataacactgc tggcttctat 2580
tgtgaccggt gcaaagacgg attttttgga aatcccctgg ctcccaatcc agcagacaaa 2640
tgcaaagcct gcaattgcaa tctgtatggg accatgaagc agcagagcag ctgtaacccc 2700
gtgacggggc agtgtgaatg tttgcctcac gtgactggcc aggactgtgg tgcttgtgac 2760
cctggattct acaatctgca gagtgggcaa ggctgtgaga ggtgtgactg ccatgccttg 2820
ggctccacca atgggcagtg tgacatccgc accggccagt gtgagtgcca gcccggcatc 2880
actggtcagc actgtgagcg ctgtgaggtc aaccactttg ggtttggacc tgaaggctgc 2940
aaaccctgtg actgtcatcc tgagggatct ctttcacttc agtgcaaaga tgatggtcgc 3000
tgtgaatgca gagaaggctt tgtgggaaat cgctgtgacc agtgtgaaga aaactatttc 3060
tacaatcggt cttggcctgg ctgccaggaa tgtccagctt gttaccggct ggtaaaggat 3120
aaggttgctg atcatagagt gaagctccag gaattagaga gtctcatagc aaaccttgga 3180
actggggatg agatggtgac agatcaagcc ttcgaggata gactaaagga agcagagagg 3240
gaagttatgg acctccttcg tgaggcccag gatgtcaaag atgttgacca gaatttgatg 3300
gatcgcctac agagagtgaa taacactctg tccagccaaa ttagccgttt acagaatatc 3360
cggaatacca ttgaagagac tggaaacttg gctgaacaag cgcgtgccca tgtagagaac 3420
acagagcggt tgattgaaat cgcatccaga gaacttgaga aagcaaaagt cgctgctgcc 3480
aatgtgtcag tcactcagcc agaatctaca ggggacccaa acaacatgac tcttttggca 3540
gaagaggctc gaaagcttgc tgaacgtcat aaacaggaag ctgatgacat tgttcgagtg 3600
gcaaagacag ccaatgatac gtcaactgag gcatacaacc tgcttctgag gacactggca 3660
ggagaaaatc aaacagcatt tgagattgaa gagcttaata ggaagtatga acaagcgaag 3720
aacatctcac aggatctgga aaaacaagct gcccgagtac atgaggaggc caaaagggcc 3780
ggtgacaaag ctgtggagat ctatgccagc gtggctcagc tgagcccttt ggactctgag 3840
acactggaga atgaagcaaa taacataaag atggaagctg agaatctgga acaactgatt 3900
gaccagaaat taaaagatta tgaggacctc agagaagata tgagagggaa ggaacttgaa 3960
gtcaagaacc ttctggagaa aggcaagact gaacagcaga ccgcagacca actcctagcc 4020
cgagctgatg ctgccaaggc cctcgctgaa gaagctgcaa agaagggacg ggatacctta 4080
caagaagcta atgacattct caacaacctg aaagattttg ataggcgtgt gaacgataac 4140
aagacggccg cagaggaggc actaaggaag attcctgcca tcaaccagac catcactgaa 4200
gccaatgaaa agaccagaga agcccagcag gccctgggca gtgctgcggc ggatgccaca 4260
gaggccaaga acaaggccca tgaggcggag aggatcgcga gcgctgtcca aaagaatgcc 4320
accagcacca aggcagaagc tgaaagaact tttgcagaag ttacagatct ggataatgag 4380
gtgaacaata tgttgaagca actgcaggaa gcagaaaaag agctaaagag aaaacaagat 4440
gacgctgacc aggacatgat gatggcaggg atggcttcac aggctgctca agaagccgag 4500
atcaatgcca gaaaagccaa aaactctgtt actagcctcc tcagcattat taatgacctc 4560
ttggagcagc tggggcagct ggatacagtg gacctgaata agctaaacga gattgaaggc 4620
accctaaaca aagccaaaga tgaaatgaag gtcagcgatc ttgataggaa agtgtctgac 4680
ctggagaatg aagccaagaa gcaggaggct gccatcatgg actataaccg agatatcgag 4740
gagatcatga aggacattcg caatctggag gacatcagga agaccttacc atctggctgc 4800
ttcaacaccc cgtccattga aaagccctag 4830
<210> 126
<211> 3122
<212> PRT
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元alpha-2的氨基酸序列(人)
<400> 126
Met Pro Gly Ala Ala Gly Val Leu Leu Leu Leu Leu Leu Ser Gly Gly
1 5 10 15
Leu Gly Gly Val Gln Ala Gln Arg Pro Gln Gln Gln Arg Gln Ser Gln
20 25 30
Ala His Gln Gln Arg Gly Leu Phe Pro Ala Val Leu Asn Leu Ala Ser
35 40 45
Asn Ala Leu Ile Thr Thr Asn Ala Thr Cys Gly Glu Lys Gly Pro Glu
50 55 60
Met Tyr Cys Lys Leu Val Glu His Val Pro Gly Gln Pro Val Arg Asn
65 70 75 80
Pro Gln Cys Arg Ile Cys Asn Gln Asn Ser Ser Asn Pro Asn Gln Arg
85 90 95
His Pro Ile Thr Asn Ala Ile Asp Gly Lys Asn Thr Trp Trp Gln Ser
100 105 110
Pro Ser Ile Lys Asn Gly Ile Glu Tyr His Tyr Val Thr Ile Thr Leu
115 120 125
Asp Leu Gln Gln Val Phe Gln Ile Ala Tyr Val Ile Val Lys Ala Ala
130 135 140
Asn Ser Pro Arg Pro Gly Asn Trp Ile Leu Glu Arg Ser Leu Asp Asp
145 150 155 160
Val Glu Tyr Lys Pro Trp Gln Tyr His Ala Val Thr Asp Thr Glu Cys
165 170 175
Leu Thr Leu Tyr Asn Ile Tyr Pro Arg Thr Gly Pro Pro Ser Tyr Ala
180 185 190
Lys Asp Asp Glu Val Ile Cys Thr Ser Phe Tyr Ser Lys Ile His Pro
195 200 205
Leu Glu Asn Gly Glu Ile His Ile Ser Leu Ile Asn Gly Arg Pro Ser
210 215 220
Ala Asp Asp Pro Ser Pro Glu Leu Leu Glu Phe Thr Ser Ala Arg Tyr
225 230 235 240
Ile Arg Leu Arg Phe Gln Arg Ile Arg Thr Leu Asn Ala Asp Leu Met
245 250 255
Met Phe Ala His Lys Asp Pro Arg Glu Ile Asp Pro Ile Val Thr Arg
260 265 270
Arg Tyr Tyr Tyr Ser Val Lys Asp Ile Ser Val Gly Gly Met Cys Ile
275 280 285
Cys Tyr Gly His Ala Arg Ala Cys Pro Leu Asp Pro Ala Thr Asn Lys
290 295 300
Ser Arg Cys Glu Cys Glu His Asn Thr Cys Gly Asp Ser Cys Asp Gln
305 310 315 320
Cys Cys Pro Gly Phe His Gln Lys Pro Trp Arg Ala Gly Thr Phe Leu
325 330 335
Thr Lys Thr Glu Cys Glu Ala Cys Asn Cys His Gly Lys Ala Glu Glu
340 345 350
Cys Tyr Tyr Asp Glu Asn Val Ala Arg Arg Asn Leu Ser Leu Asn Ile
355 360 365
Arg Gly Lys Tyr Ile Gly Gly Gly Val Cys Ile Asn Cys Thr Gln Asn
370 375 380
Thr Ala Gly Ile Asn Cys Glu Thr Cys Thr Asp Gly Phe Phe Arg Pro
385 390 395 400
Lys Gly Val Ser Pro Asn Tyr Pro Arg Pro Cys Gln Pro Cys His Cys
405 410 415
Asp Pro Ile Gly Ser Leu Asn Glu Val Cys Val Lys Asp Glu Lys His
420 425 430
Ala Arg Arg Gly Leu Ala Pro Gly Ser Cys His Cys Lys Thr Gly Phe
435 440 445
Gly Gly Val Ser Cys Asp Arg Cys Ala Arg Gly Tyr Thr Gly Tyr Pro
450 455 460
Asp Cys Lys Ala Cys Asn Cys Ser Gly Leu Gly Ser Lys Asn Glu Asp
465 470 475 480
Pro Cys Phe Gly Pro Cys Ile Cys Lys Glu Asn Val Glu Gly Gly Asp
485 490 495
Cys Ser Arg Cys Lys Ser Gly Phe Phe Asn Leu Gln Glu Asp Asn Trp
500 505 510
Lys Gly Cys Asp Glu Cys Phe Cys Ser Gly Val Ser Asn Arg Cys Gln
515 520 525
Ser Ser Tyr Trp Thr Tyr Gly Lys Ile Gln Asp Met Ser Gly Trp Tyr
530 535 540
Leu Thr Asp Leu Pro Gly Arg Ile Arg Val Ala Pro Gln Gln Asp Asp
545 550 555 560
Leu Asp Ser Pro Gln Gln Ile Ser Ile Ser Asn Ala Glu Ala Arg Gln
565 570 575
Ala Leu Pro His Ser Tyr Tyr Trp Ser Ala Pro Ala Pro Tyr Leu Gly
580 585 590
Asn Lys Leu Pro Ala Val Gly Gly Gln Leu Thr Phe Thr Ile Ser Tyr
595 600 605
Asp Leu Glu Glu Glu Glu Glu Asp Thr Glu Arg Val Leu Gln Leu Met
610 615 620
Ile Ile Leu Glu Gly Asn Asp Leu Ser Ile Ser Thr Ala Gln Asp Glu
625 630 635 640
Val Tyr Leu His Pro Ser Glu Glu His Thr Asn Val Leu Leu Leu Lys
645 650 655
Glu Glu Ser Phe Thr Ile His Gly Thr His Phe Pro Val Arg Arg Lys
660 665 670
Glu Phe Met Thr Val Leu Ala Asn Leu Lys Arg Val Leu Leu Gln Ile
675 680 685
Thr Tyr Ser Phe Gly Met Asp Ala Ile Phe Arg Leu Ser Ser Val Asn
690 695 700
Leu Glu Ser Ala Val Ser Tyr Pro Thr Asp Gly Ser Ile Ala Ala Ala
705 710 715 720
Val Glu Val Cys Gln Cys Pro Pro Gly Tyr Thr Gly Ser Ser Cys Glu
725 730 735
Ser Cys Trp Pro Arg His Arg Arg Val Asn Gly Thr Ile Phe Gly Gly
740 745 750
Ile Cys Glu Pro Cys Gln Cys Phe Gly His Ala Glu Ser Cys Asp Asp
755 760 765
Val Thr Gly Glu Cys Leu Asn Cys Lys Asp His Thr Gly Gly Pro Tyr
770 775 780
Cys Asp Lys Cys Leu Pro Gly Phe Tyr Gly Glu Pro Thr Lys Gly Thr
785 790 795 800
Ser Glu Asp Cys Gln Pro Cys Ala Cys Pro Leu Asn Ile Pro Ser Asn
805 810 815
Asn Phe Ser Pro Thr Cys His Leu Asp Arg Ser Leu Gly Leu Ile Cys
820 825 830
Asp Gly Cys Pro Val Gly Tyr Thr Gly Pro Arg Cys Glu Arg Cys Ala
835 840 845
Glu Gly Tyr Phe Gly Gln Pro Ser Val Pro Gly Gly Ser Cys Gln Pro
850 855 860
Cys Gln Cys Asn Asp Asn Leu Asp Phe Ser Ile Pro Gly Ser Cys Asp
865 870 875 880
Ser Leu Ser Gly Ser Cys Leu Ile Cys Lys Pro Gly Thr Thr Gly Arg
885 890 895
Tyr Cys Glu Leu Cys Ala Asp Gly Tyr Phe Gly Asp Ala Val Asp Ala
900 905 910
Lys Asn Cys Gln Pro Cys Arg Cys Asn Ala Gly Gly Ser Phe Ser Glu
915 920 925
Val Cys His Ser Gln Thr Gly Gln Cys Glu Cys Arg Ala Asn Val Gln
930 935 940
Gly Gln Arg Cys Asp Lys Cys Lys Ala Gly Thr Phe Gly Leu Gln Ser
945 950 955 960
Ala Arg Gly Cys Val Pro Cys Asn Cys Asn Ser Phe Gly Ser Lys Ser
965 970 975
Phe Asp Cys Glu Glu Ser Gly Gln Cys Trp Cys Gln Pro Gly Val Thr
980 985 990
Gly Lys Lys Cys Asp Arg Cys Ala His Gly Tyr Phe Asn Phe Gln Glu
995 1000 1005
Gly Gly Cys Thr Ala Cys Glu Cys Ser His Leu Gly Asn Asn Cys Asp
1010 1015 1020
Pro Lys Thr Gly Arg Cys Ile Cys Pro Pro Asn Thr Ile Gly Glu Lys
1025 1030 1035 1040
Cys Ser Lys Cys Ala Pro Asn Thr Trp Gly His Ser Ile Thr Thr Gly
1045 1050 1055
Cys Lys Ala Cys Asn Cys Ser Thr Val Gly Ser Leu Asp Phe Gln Cys
1060 1065 1070
Asn Val Asn Thr Gly Gln Cys Asn Cys His Pro Lys Phe Ser Gly Ala
1075 1080 1085
Lys Cys Thr Glu Cys Ser Arg Gly His Trp Asn Tyr Pro Arg Cys Asn
1090 1095 1100
Leu Cys Asp Cys Phe Leu Pro Gly Thr Asp Ala Thr Thr Cys Asp Ser
1105 1110 1115 1120
Glu Thr Lys Lys Cys Ser Cys Ser Asp Gln Thr Gly Gln Cys Thr Cys
1125 1130 1135
Lys Val Asn Val Glu Gly Ile His Cys Asp Arg Cys Arg Pro Gly Lys
1140 1145 1150
Phe Gly Leu Asp Ala Lys Asn Pro Leu Gly Cys Ser Ser Cys Tyr Cys
1155 1160 1165
Phe Gly Thr Thr Thr Gln Cys Ser Glu Ala Lys Gly Leu Ile Arg Thr
1170 1175 1180
Trp Val Thr Leu Lys Ala Glu Gln Thr Ile Leu Pro Leu Val Asp Glu
1185 1190 1195 1200
Ala Leu Gln His Thr Thr Thr Lys Gly Ile Val Phe Gln His Pro Glu
1205 1210 1215
Ile Val Ala His Met Asp Leu Met Arg Glu Asp Leu His Leu Glu Pro
1220 1225 1230
Phe Tyr Trp Lys Leu Pro Glu Gln Phe Glu Gly Lys Lys Leu Met Ala
1235 1240 1245
Tyr Gly Gly Lys Leu Lys Tyr Ala Ile Tyr Phe Glu Ala Arg Glu Glu
1250 1255 1260
Thr Gly Phe Ser Thr Tyr Asn Pro Gln Val Ile Ile Arg Gly Gly Thr
1265 1270 1275 1280
Pro Thr His Ala Arg Ile Ile Val Arg His Met Ala Ala Pro Leu Ile
1285 1290 1295
Gly Gln Leu Thr Arg His Glu Ile Glu Met Thr Glu Lys Glu Trp Lys
1300 1305 1310
Tyr Tyr Gly Asp Asp Pro Arg Val His Arg Thr Val Thr Arg Glu Asp
1315 1320 1325
Phe Leu Asp Ile Leu Tyr Asp Ile His Tyr Ile Leu Ile Lys Ala Thr
1330 1335 1340
Tyr Gly Asn Phe Met Arg Gln Ser Arg Ile Ser Glu Ile Ser Met Glu
1345 1350 1355 1360
Val Ala Glu Gln Gly Arg Gly Thr Thr Met Thr Pro Pro Ala Asp Leu
1365 1370 1375
Ile Glu Lys Cys Asp Cys Pro Leu Gly Tyr Ser Gly Leu Ser Cys Glu
1380 1385 1390
Ala Cys Leu Pro Gly Phe Tyr Arg Leu Arg Ser Gln Pro Gly Gly Arg
1395 1400 1405
Thr Pro Gly Pro Thr Leu Gly Thr Cys Val Pro Cys Gln Cys Asn Gly
1410 1415 1420
His Ser Ser Leu Cys Asp Pro Glu Thr Ser Ile Cys Gln Asn Cys Gln
1425 1430 1435 1440
His His Thr Ala Gly Asp Phe Cys Glu Arg Cys Ala Leu Gly Tyr Tyr
1445 1450 1455
Gly Ile Val Lys Gly Leu Pro Asn Asp Cys Gln Gln Cys Ala Cys Pro
1460 1465 1470
Leu Ile Ser Ser Ser Asn Asn Phe Ser Pro Ser Cys Val Ala Glu Gly
1475 1480 1485
Leu Asp Asp Tyr Arg Cys Thr Ala Cys Pro Arg Gly Tyr Glu Gly Gln
1490 1495 1500
Tyr Cys Glu Arg Cys Ala Pro Gly Tyr Thr Gly Ser Pro Gly Asn Pro
1505 1510 1515 1520
Gly Gly Ser Cys Gln Glu Cys Glu Cys Asp Pro Tyr Gly Ser Leu Pro
1525 1530 1535
Val Pro Cys Asp Pro Val Thr Gly Phe Cys Thr Cys Arg Pro Gly Ala
1540 1545 1550
Thr Gly Arg Lys Cys Asp Gly Cys Lys His Trp His Ala Arg Glu Gly
1555 1560 1565
Trp Glu Cys Val Phe Cys Gly Asp Glu Cys Thr Gly Leu Leu Leu Gly
1570 1575 1580
Asp Leu Ala Arg Leu Glu Gln Met Val Met Ser Ile Asn Leu Thr Gly
1585 1590 1595 1600
Pro Leu Pro Ala Pro Tyr Lys Met Leu Tyr Gly Leu Glu Asn Met Thr
1605 1610 1615
Gln Glu Leu Lys His Leu Leu Ser Pro Gln Arg Ala Pro Glu Arg Leu
1620 1625 1630
Ile Gln Leu Ala Glu Gly Asn Leu Asn Thr Leu Val Thr Glu Met Asn
1635 1640 1645
Glu Leu Leu Thr Arg Ala Thr Lys Val Thr Ala Asp Gly Glu Gln Thr
1650 1655 1660
Gly Gln Asp Ala Glu Arg Thr Asn Thr Arg Ala Lys Ser Leu Gly Glu
1665 1670 1675 1680
Phe Ile Lys Glu Leu Ala Arg Asp Ala Glu Ala Val Asn Glu Lys Ala
1685 1690 1695
Ile Lys Leu Asn Glu Thr Leu Gly Thr Arg Asp Glu Ala Phe Glu Arg
1700 1705 1710
Asn Leu Glu Gly Leu Gln Lys Glu Ile Asp Gln Met Ile Lys Glu Leu
1715 1720 1725
Arg Arg Lys Asn Leu Glu Thr Gln Lys Glu Ile Ala Glu Asp Glu Leu
1730 1735 1740
Val Ala Ala Glu Ala Leu Leu Lys Lys Val Lys Lys Leu Phe Gly Glu
1745 1750 1755 1760
Ser Arg Gly Glu Asn Glu Glu Met Glu Lys Asp Leu Arg Glu Lys Leu
1765 1770 1775
Ala Asp Tyr Lys Asn Lys Val Asp Asp Ala Trp Asp Leu Leu Arg Glu
1780 1785 1790
Ala Thr Asp Lys Ile Arg Glu Ala Asn Arg Leu Phe Ala Val Asn Gln
1795 1800 1805
Lys Asn Met Thr Ala Leu Glu Lys Lys Lys Glu Ala Val Glu Ser Gly
1810 1815 1820
Lys Arg Gln Ile Glu Asn Thr Leu Lys Glu Gly Asn Asp Ile Leu Asp
1825 1830 1835 1840
Glu Ala Asn Arg Leu Ala Asp Glu Ile Asn Ser Ile Ile Asp Tyr Val
1845 1850 1855
Glu Asp Ile Gln Thr Lys Leu Pro Pro Met Ser Glu Glu Leu Asn Asp
1860 1865 1870
Lys Ile Asp Asp Leu Ser Gln Glu Ile Lys Asp Arg Lys Leu Ala Glu
1875 1880 1885
Lys Val Ser Gln Ala Glu Ser His Ala Ala Gln Leu Asn Asp Ser Ser
1890 1895 1900
Ala Val Leu Asp Gly Ile Leu Asp Glu Ala Lys Asn Ile Ser Phe Asn
1905 1910 1915 1920
Ala Thr Ala Ala Phe Lys Ala Tyr Ser Asn Ile Lys Asp Tyr Ile Asp
1925 1930 1935
Glu Ala Glu Lys Val Ala Lys Glu Ala Lys Asp Leu Ala His Glu Ala
1940 1945 1950
Thr Lys Leu Ala Thr Gly Pro Arg Gly Leu Leu Lys Glu Asp Ala Lys
1955 1960 1965
Gly Cys Leu Gln Lys Ser Phe Arg Ile Leu Asn Glu Ala Lys Lys Leu
1970 1975 1980
Ala Asn Asp Val Lys Glu Asn Glu Asp His Leu Asn Gly Leu Lys Thr
1985 1990 1995 2000
Arg Ile Glu Asn Ala Asp Ala Arg Asn Gly Asp Leu Leu Arg Thr Leu
2005 2010 2015
Asn Asp Thr Leu Gly Lys Leu Ser Ala Ile Pro Asn Asp Thr Ala Ala
2020 2025 2030
Lys Leu Gln Ala Val Lys Asp Lys Ala Arg Gln Ala Asn Asp Thr Ala
2035 2040 2045
Lys Asp Val Leu Ala Gln Ile Thr Glu Leu His Gln Asn Leu Asp Gly
2050 2055 2060
Leu Lys Lys Asn Tyr Asn Lys Leu Ala Asp Ser Val Ala Lys Thr Asn
2065 2070 2075 2080
Ala Val Val Lys Asp Pro Ser Lys Asn Lys Ile Ile Ala Asp Ala Asp
2085 2090 2095
Ala Thr Val Lys Asn Leu Glu Gln Glu Ala Asp Arg Leu Ile Asp Lys
2100 2105 2110
Leu Lys Pro Ile Lys Glu Leu Glu Asp Asn Leu Lys Lys Asn Ile Ser
2115 2120 2125
Glu Ile Lys Glu Leu Ile Asn Gln Ala Arg Lys Gln Ala Asn Ser Ile
2130 2135 2140
Lys Val Ser Val Ser Ser Gly Gly Asp Cys Ile Arg Thr Tyr Lys Pro
2145 2150 2155 2160
Glu Ile Lys Lys Gly Ser Tyr Asn Asn Ile Val Val Asn Val Lys Thr
2165 2170 2175
Ala Val Ala Asp Asn Leu Leu Phe Tyr Leu Gly Ser Ala Lys Phe Ile
2180 2185 2190
Asp Phe Leu Ala Ile Glu Met Arg Lys Gly Lys Val Ser Phe Leu Trp
2195 2200 2205
Asp Val Gly Ser Gly Val Gly Arg Val Glu Tyr Pro Asp Leu Thr Ile
2210 2215 2220
Asp Asp Ser Tyr Trp Tyr Arg Ile Val Ala Ser Arg Thr Gly Arg Asn
2225 2230 2235 2240
Gly Thr Ile Ser Val Arg Ala Leu Asp Gly Pro Lys Ala Ser Ile Val
2245 2250 2255
Pro Ser Thr His His Ser Thr Ser Pro Pro Gly Tyr Thr Ile Leu Asp
2260 2265 2270
Val Asp Ala Asn Ala Met Leu Phe Val Gly Gly Leu Thr Gly Lys Leu
2275 2280 2285
Lys Lys Ala Asp Ala Val Arg Val Ile Thr Phe Thr Gly Cys Met Gly
2290 2295 2300
Glu Thr Tyr Phe Asp Asn Lys Pro Ile Gly Leu Trp Asn Phe Arg Glu
2305 2310 2315 2320
Lys Glu Gly Asp Cys Lys Gly Cys Thr Val Ser Pro Gln Val Glu Asp
2325 2330 2335
Ser Glu Gly Thr Ile Gln Phe Asp Gly Glu Gly Tyr Ala Leu Val Ser
2340 2345 2350
Arg Pro Ile Arg Trp Tyr Pro Asn Ile Ser Thr Val Met Phe Lys Phe
2355 2360 2365
Arg Thr Phe Ser Ser Ser Ala Leu Leu Met Tyr Leu Ala Thr Arg Asp
2370 2375 2380
Leu Arg Asp Phe Met Ser Val Glu Leu Thr Asp Gly His Ile Lys Val
2385 2390 2395 2400
Ser Tyr Asp Leu Gly Ser Gly Met Ala Ser Val Val Ser Asn Gln Asn
2405 2410 2415
His Asn Asp Gly Lys Trp Lys Ser Phe Thr Leu Ser Arg Ile Gln Lys
2420 2425 2430
Gln Ala Asn Ile Ser Ile Val Asp Ile Asp Thr Asn Gln Glu Glu Asn
2435 2440 2445
Ile Ala Thr Ser Ser Ser Gly Asn Asn Phe Gly Leu Asp Leu Lys Ala
2450 2455 2460
Asp Asp Lys Ile Tyr Phe Gly Gly Leu Pro Thr Leu Arg Asn Leu Ser
2465 2470 2475 2480
Met Lys Ala Arg Pro Glu Val Asn Leu Lys Lys Tyr Ser Gly Cys Leu
2485 2490 2495
Lys Asp Ile Glu Ile Ser Arg Thr Pro Tyr Asn Ile Leu Ser Ser Pro
2500 2505 2510
Asp Tyr Val Gly Val Thr Lys Gly Cys Ser Leu Glu Asn Val Tyr Thr
2515 2520 2525
Val Ser Phe Pro Lys Pro Gly Phe Val Glu Leu Ser Pro Val Pro Ile
2530 2535 2540
Asp Val Gly Thr Glu Ile Asn Leu Ser Phe Ser Thr Lys Asn Glu Ser
2545 2550 2555 2560
Gly Ile Ile Leu Leu Gly Ser Gly Gly Thr Pro Ala Pro Pro Arg Arg
2565 2570 2575
Lys Arg Arg Gln Thr Gly Gln Ala Tyr Tyr Ala Ile Leu Leu Asn Arg
2580 2585 2590
Gly Arg Leu Glu Val His Leu Ser Thr Gly Ala Arg Thr Met Arg Lys
2595 2600 2605
Ile Val Ile Arg Pro Glu Pro Asn Leu Phe His Asp Gly Arg Glu His
2610 2615 2620
Ser Val His Val Glu Arg Thr Arg Gly Ile Phe Thr Val Gln Val Asp
2625 2630 2635 2640
Glu Asn Arg Arg Tyr Met Gln Asn Leu Thr Val Glu Gln Pro Ile Glu
2645 2650 2655
Val Lys Lys Leu Phe Val Gly Gly Ala Pro Pro Glu Phe Gln Pro Ser
2660 2665 2670
Pro Leu Arg Asn Ile Pro Pro Phe Glu Gly Cys Ile Trp Asn Leu Val
2675 2680 2685
Ile Asn Ser Val Pro Met Asp Phe Ala Arg Pro Val Ser Phe Lys Asn
2690 2695 2700
Ala Asp Ile Gly Arg Cys Ala His Gln Lys Leu Arg Glu Asp Glu Asp
2705 2710 2715 2720
Gly Ala Ala Pro Ala Glu Ile Val Ile Gln Pro Glu Pro Val Pro Thr
2725 2730 2735
Pro Ala Phe Pro Thr Pro Thr Pro Val Leu Thr His Gly Pro Cys Ala
2740 2745 2750
Ala Glu Ser Glu Pro Ala Leu Leu Ile Gly Ser Lys Gln Phe Gly Leu
2755 2760 2765
Ser Arg Asn Ser His Ile Ala Ile Ala Phe Asp Asp Thr Lys Val Lys
2770 2775 2780
Asn Arg Leu Thr Ile Glu Leu Glu Val Arg Thr Glu Ala Glu Ser Gly
2785 2790 2795 2800
Leu Leu Phe Tyr Met Ala Arg Ile Asn His Ala Asp Phe Ala Thr Val
2805 2810 2815
Gln Leu Arg Asn Gly Leu Pro Tyr Phe Ser Tyr Asp Leu Gly Ser Gly
2820 2825 2830
Asp Thr His Thr Met Ile Pro Thr Lys Ile Asn Asp Gly Gln Trp His
2835 2840 2845
Lys Ile Lys Ile Met Arg Ser Lys Gln Glu Gly Ile Leu Tyr Val Asp
2850 2855 2860
Gly Ala Ser Asn Arg Thr Ile Ser Pro Lys Lys Ala Asp Ile Leu Asp
2865 2870 2875 2880
Val Val Gly Met Leu Tyr Val Gly Gly Leu Pro Ile Asn Tyr Thr Thr
2885 2890 2895
Arg Arg Ile Gly Pro Val Thr Tyr Ser Ile Asp Gly Cys Val Arg Asn
2900 2905 2910
Leu His Met Ala Glu Ala Pro Ala Asp Leu Glu Gln Pro Thr Ser Ser
2915 2920 2925
Phe His Val Gly Thr Cys Phe Ala Asn Ala Gln Arg Gly Thr Tyr Phe
2930 2935 2940
Asp Gly Thr Gly Phe Ala Lys Ala Val Gly Gly Phe Lys Val Gly Leu
2945 2950 2955 2960
Asp Leu Leu Val Glu Phe Glu Phe Arg Thr Thr Thr Thr Thr Gly Val
2965 2970 2975
Leu Leu Gly Ile Ser Ser Gln Lys Met Asp Gly Met Gly Ile Glu Met
2980 2985 2990
Ile Asp Glu Lys Leu Met Phe His Val Asp Asn Gly Ala Gly Arg Phe
2995 3000 3005
Thr Ala Val Tyr Asp Ala Gly Val Pro Gly His Leu Cys Asp Gly Gln
3010 3015 3020
Trp His Lys Val Thr Ala Asn Lys Ile Lys His Arg Ile Glu Leu Thr
3025 3030 3035 3040
Val Asp Gly Asn Gln Val Glu Ala Gln Ser Pro Asn Pro Ala Ser Thr
3045 3050 3055
Ser Ala Asp Thr Asn Asp Pro Val Phe Val Gly Gly Phe Pro Asp Asp
3060 3065 3070
Leu Lys Gln Phe Gly Leu Thr Thr Ser Ile Pro Phe Arg Gly Cys Ile
3075 3080 3085
Arg Ser Leu Lys Leu Thr Lys Gly Thr Gly Lys Pro Leu Glu Val Asn
3090 3095 3100
Phe Ala Lys Ala Leu Glu Leu Arg Gly Val Gln Pro Val Ser Cys Pro
3105 3110 3115 3120
Ala Asn
<210> 127
<211> 3695
<212> PRT
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元alpha-5的氨基酸序列(人)
<400> 127
Met Ala Lys Arg Leu Cys Ala Gly Ser Ala Leu Cys Val Arg Gly Pro
1 5 10 15
Arg Gly Pro Ala Pro Leu Leu Leu Val Gly Leu Ala Leu Leu Gly Ala
20 25 30
Ala Arg Ala Arg Glu Glu Ala Gly Gly Gly Phe Ser Leu His Pro Pro
35 40 45
Tyr Phe Asn Leu Ala Glu Gly Ala Arg Ile Ala Ala Ser Ala Thr Cys
50 55 60
Gly Glu Glu Ala Pro Ala Arg Gly Ser Pro Arg Pro Thr Glu Asp Leu
65 70 75 80
Tyr Cys Lys Leu Val Gly Gly Pro Val Ala Gly Gly Asp Pro Asn Gln
85 90 95
Thr Ile Arg Gly Gln Tyr Cys Asp Ile Cys Thr Ala Ala Asn Ser Asn
100 105 110
Lys Ala His Pro Ala Ser Asn Ala Ile Asp Gly Thr Glu Arg Trp Trp
115 120 125
Gln Ser Pro Pro Leu Ser Arg Gly Leu Glu Tyr Asn Glu Val Asn Val
130 135 140
Thr Leu Asp Leu Gly Gln Val Phe His Val Ala Tyr Val Leu Ile Lys
145 150 155 160
Phe Ala Asn Ser Pro Arg Pro Asp Leu Trp Val Leu Glu Arg Ser Met
165 170 175
Asp Phe Gly Arg Thr Tyr Gln Pro Trp Gln Phe Phe Ala Ser Ser Lys
180 185 190
Arg Asp Cys Leu Glu Arg Phe Gly Pro Gln Thr Leu Glu Arg Ile Thr
195 200 205
Arg Asp Asp Ala Ala Ile Cys Thr Thr Glu Tyr Ser Arg Ile Val Pro
210 215 220
Leu Glu Asn Gly Glu Ile Val Val Ser Leu Val Asn Gly Arg Pro Gly
225 230 235 240
Ala Met Asn Phe Ser Tyr Ser Pro Leu Leu Arg Glu Phe Thr Lys Ala
245 250 255
Thr Asn Val Arg Leu Arg Phe Leu Arg Thr Asn Thr Leu Leu Gly His
260 265 270
Leu Met Gly Lys Ala Leu Arg Asp Pro Thr Val Thr Arg Arg Tyr Tyr
275 280 285
Tyr Ser Ile Lys Asp Ile Ser Ile Gly Gly Arg Cys Val Cys His Gly
290 295 300
His Ala Asp Ala Cys Asp Ala Lys Asp Pro Thr Asp Pro Phe Arg Leu
305 310 315 320
Gln Cys Thr Cys Gln His Asn Thr Cys Gly Gly Thr Cys Asp Arg Cys
325 330 335
Cys Pro Gly Phe Asn Gln Gln Pro Trp Lys Pro Ala Thr Ala Asn Ser
340 345 350
Ala Asn Glu Cys Gln Ser Cys Asn Cys Tyr Gly His Ala Thr Asp Cys
355 360 365
Tyr Tyr Asp Pro Glu Val Asp Arg Arg Arg Ala Ser Gln Ser Leu Asp
370 375 380
Gly Thr Tyr Gln Gly Gly Gly Val Cys Ile Asp Cys Gln His His Thr
385 390 395 400
Thr Gly Val Asn Cys Glu Arg Cys Leu Pro Gly Phe Tyr Arg Ser Pro
405 410 415
Asn His Pro Leu Asp Ser Pro His Val Cys Arg Arg Cys Asn Cys Glu
420 425 430
Ser Asp Phe Thr Asp Gly Thr Cys Glu Asp Leu Thr Gly Arg Cys Tyr
435 440 445
Cys Arg Pro Asn Phe Ser Gly Glu Arg Cys Asp Val Cys Ala Glu Gly
450 455 460
Phe Thr Gly Phe Pro Ser Cys Tyr Pro Thr Pro Ser Ser Ser Asn Asp
465 470 475 480
Thr Arg Glu Gln Val Leu Pro Ala Gly Gln Ile Val Asn Cys Asp Cys
485 490 495
Ser Ala Ala Gly Thr Gln Gly Asn Ala Cys Arg Lys Asp Pro Arg Val
500 505 510
Gly Arg Cys Leu Cys Lys Pro Asn Phe Gln Gly Thr His Cys Glu Leu
515 520 525
Cys Ala Pro Gly Phe Tyr Gly Pro Gly Cys Gln Pro Cys Gln Cys Ser
530 535 540
Ser Pro Gly Val Ala Asp Asp Arg Cys Asp Pro Asp Thr Gly Gln Cys
545 550 555 560
Arg Cys Arg Val Gly Phe Glu Gly Ala Thr Cys Asp Arg Cys Ala Pro
565 570 575
Gly Tyr Phe His Phe Pro Leu Cys Gln Leu Cys Gly Cys Ser Pro Ala
580 585 590
Gly Thr Leu Pro Glu Gly Cys Asp Glu Ala Gly Arg Cys Leu Cys Gln
595 600 605
Pro Glu Phe Ala Gly Pro His Cys Asp Arg Cys Arg Pro Gly Tyr His
610 615 620
Gly Phe Pro Asn Cys Gln Ala Cys Thr Cys Asp Pro Arg Gly Ala Leu
625 630 635 640
Asp Gln Leu Cys Gly Ala Gly Gly Leu Cys Arg Cys Arg Pro Gly Tyr
645 650 655
Thr Gly Thr Ala Cys Gln Glu Cys Ser Pro Gly Phe His Gly Phe Pro
660 665 670
Ser Cys Val Pro Cys His Cys Ser Ala Glu Gly Ser Leu His Ala Ala
675 680 685
Cys Asp Pro Arg Ser Gly Gln Cys Ser Cys Arg Pro Arg Val Thr Gly
690 695 700
Leu Arg Cys Asp Thr Cys Val Pro Gly Ala Tyr Asn Phe Pro Tyr Cys
705 710 715 720
Glu Ala Gly Ser Cys His Pro Ala Gly Leu Ala Pro Val Asp Pro Ala
725 730 735
Leu Pro Glu Ala Gln Val Pro Cys Met Cys Arg Ala His Val Glu Gly
740 745 750
Pro Ser Cys Asp Arg Cys Lys Pro Gly Phe Trp Gly Leu Ser Pro Ser
755 760 765
Asn Pro Glu Gly Cys Thr Arg Cys Ser Cys Asp Leu Arg Gly Thr Leu
770 775 780
Gly Gly Val Ala Glu Cys Gln Pro Gly Thr Gly Gln Cys Phe Cys Lys
785 790 795 800
Pro His Val Cys Gly Gln Ala Cys Ala Ser Cys Lys Asp Gly Phe Phe
805 810 815
Gly Leu Asp Gln Ala Asp Tyr Phe Gly Cys Arg Ser Cys Arg Cys Asp
820 825 830
Ile Gly Gly Ala Leu Gly Gln Ser Cys Glu Pro Arg Thr Gly Val Cys
835 840 845
Arg Cys Arg Pro Asn Thr Gln Gly Pro Thr Cys Ser Glu Pro Ala Arg
850 855 860
Asp His Tyr Leu Pro Asp Leu His His Leu Arg Leu Glu Leu Glu Glu
865 870 875 880
Ala Ala Thr Pro Glu Gly His Ala Val Arg Phe Gly Phe Asn Pro Leu
885 890 895
Glu Phe Glu Asn Phe Ser Trp Arg Gly Tyr Ala Gln Met Ala Pro Val
900 905 910
Gln Pro Arg Ile Val Ala Arg Leu Asn Leu Thr Ser Pro Asp Leu Phe
915 920 925
Trp Leu Val Phe Arg Tyr Val Asn Arg Gly Ala Met Ser Val Ser Gly
930 935 940
Arg Val Ser Val Arg Glu Glu Gly Arg Ser Ala Thr Cys Ala Asn Cys
945 950 955 960
Thr Ala Gln Ser Gln Pro Val Ala Phe Pro Pro Ser Thr Glu Pro Ala
965 970 975
Phe Ile Thr Val Pro Gln Arg Gly Phe Gly Glu Pro Phe Val Leu Asn
980 985 990
Pro Gly Thr Trp Ala Leu Arg Val Glu Ala Glu Gly Val Leu Leu Asp
995 1000 1005
Tyr Val Val Leu Leu Pro Ser Ala Tyr Tyr Glu Ala Ala Leu Leu Gln
1010 1015 1020
Leu Arg Val Thr Glu Ala Cys Thr Tyr Arg Pro Ser Ala Gln Gln Ser
1025 1030 1035 1040
Gly Asp Asn Cys Leu Leu Tyr Thr His Leu Pro Leu Asp Gly Phe Pro
1045 1050 1055
Ser Ala Ala Gly Leu Glu Ala Leu Cys Arg Gln Asp Asn Ser Leu Pro
1060 1065 1070
Arg Pro Cys Pro Thr Glu Gln Leu Ser Pro Ser His Pro Pro Leu Ile
1075 1080 1085
Thr Cys Thr Gly Ser Asp Val Asp Val Gln Leu Gln Val Ala Val Pro
1090 1095 1100
Gln Pro Gly Arg Tyr Ala Leu Val Val Glu Tyr Ala Asn Glu Asp Ala
1105 1110 1115 1120
Arg Gln Glu Val Gly Val Ala Val His Thr Pro Gln Arg Ala Pro Gln
1125 1130 1135
Gln Gly Leu Leu Ser Leu His Pro Cys Leu Tyr Ser Thr Leu Cys Arg
1140 1145 1150
Gly Thr Ala Arg Asp Thr Gln Asp His Leu Ala Val Phe His Leu Asp
1155 1160 1165
Ser Glu Ala Ser Val Arg Leu Thr Ala Glu Gln Ala Arg Phe Phe Leu
1170 1175 1180
His Gly Val Thr Leu Val Pro Ile Glu Glu Phe Ser Pro Glu Phe Val
1185 1190 1195 1200
Glu Pro Arg Val Ser Cys Ile Ser Ser His Gly Ala Phe Gly Pro Asn
1205 1210 1215
Ser Ala Ala Cys Leu Pro Ser Arg Phe Pro Lys Pro Pro Gln Pro Ile
1220 1225 1230
Ile Leu Arg Asp Cys Gln Val Ile Pro Leu Pro Pro Gly Leu Pro Leu
1235 1240 1245
Thr His Ala Gln Asp Leu Thr Pro Ala Met Ser Pro Ala Gly Pro Arg
1250 1255 1260
Pro Arg Pro Pro Thr Ala Val Asp Pro Asp Ala Glu Pro Thr Leu Leu
1265 1270 1275 1280
Arg Glu Pro Gln Ala Thr Val Val Phe Thr Thr His Val Pro Thr Leu
1285 1290 1295
Gly Arg Tyr Ala Phe Leu Leu His Gly Tyr Gln Pro Ala His Pro Thr
1300 1305 1310
Phe Pro Val Glu Val Leu Ile Asn Ala Gly Arg Val Trp Gln Gly His
1315 1320 1325
Ala Asn Ala Ser Phe Cys Pro His Gly Tyr Gly Cys Arg Thr Leu Val
1330 1335 1340
Val Cys Glu Gly Gln Ala Leu Leu Asp Val Thr His Ser Glu Leu Thr
1345 1350 1355 1360
Val Thr Val Arg Val Pro Lys Gly Arg Trp Leu Trp Leu Asp Tyr Val
1365 1370 1375
Leu Val Val Pro Glu Asn Val Tyr Ser Phe Gly Tyr Leu Arg Glu Glu
1380 1385 1390
Pro Leu Asp Lys Ser Tyr Asp Phe Ile Ser His Cys Ala Ala Gln Gly
1395 1400 1405
Tyr His Ile Ser Pro Ser Ser Ser Ser Leu Phe Cys Arg Asn Ala Ala
1410 1415 1420
Ala Ser Leu Ser Leu Phe Tyr Asn Asn Gly Ala Arg Pro Cys Gly Cys
1425 1430 1435 1440
His Glu Val Gly Ala Thr Gly Pro Thr Cys Glu Pro Phe Gly Gly Gln
1445 1450 1455
Cys Pro Cys His Ala His Val Ile Gly Arg Asp Cys Ser Arg Cys Ala
1460 1465 1470
Thr Gly Tyr Trp Gly Phe Pro Asn Cys Arg Pro Cys Asp Cys Gly Ala
1475 1480 1485
Arg Leu Cys Asp Glu Leu Thr Gly Gln Cys Ile Cys Pro Pro Arg Thr
1490 1495 1500
Ile Pro Pro Asp Cys Leu Leu Cys Gln Pro Gln Thr Phe Gly Cys His
1505 1510 1515 1520
Pro Leu Val Gly Cys Glu Glu Cys Asn Cys Ser Gly Pro Gly Ile Gln
1525 1530 1535
Glu Leu Thr Asp Pro Thr Cys Asp Thr Asp Ser Gly Gln Cys Lys Cys
1540 1545 1550
Arg Pro Asn Val Thr Gly Arg Arg Cys Asp Thr Cys Ser Pro Gly Phe
1555 1560 1565
His Gly Tyr Pro Arg Cys Arg Pro Cys Asp Cys His Glu Ala Gly Thr
1570 1575 1580
Ala Pro Gly Val Cys Asp Pro Leu Thr Gly Gln Cys Tyr Cys Lys Glu
1585 1590 1595 1600
Asn Val Gln Gly Pro Lys Cys Asp Gln Cys Ser Leu Gly Thr Phe Ser
1605 1610 1615
Leu Asp Ala Ala Asn Pro Lys Gly Cys Thr Arg Cys Phe Cys Phe Gly
1620 1625 1630
Ala Thr Glu Arg Cys Arg Ser Ser Ser Tyr Thr Arg Gln Glu Phe Val
1635 1640 1645
Asp Met Glu Gly Trp Val Leu Leu Ser Thr Asp Arg Gln Val Val Pro
1650 1655 1660
His Glu Arg Gln Pro Gly Thr Glu Met Leu Arg Ala Asp Leu Arg His
1665 1670 1675 1680
Val Pro Glu Ala Val Pro Glu Ala Phe Pro Glu Leu Tyr Trp Gln Ala
1685 1690 1695
Pro Pro Ser Tyr Leu Gly Asp Arg Val Ser Ser Tyr Gly Gly Thr Leu
1700 1705 1710
Arg Tyr Glu Leu His Ser Glu Thr Gln Arg Gly Asp Val Phe Val Pro
1715 1720 1725
Met Glu Ser Arg Pro Asp Val Val Leu Gln Gly Asn Gln Met Ser Ile
1730 1735 1740
Thr Phe Leu Glu Pro Ala Tyr Pro Thr Pro Gly His Val His Arg Gly
1745 1750 1755 1760
Gln Leu Gln Leu Val Glu Gly Asn Phe Arg His Thr Glu Thr Arg Asn
1765 1770 1775
Thr Val Ser Arg Glu Glu Leu Met Met Val Leu Ala Ser Leu Glu Gln
1780 1785 1790
Leu Gln Ile Arg Ala Leu Phe Ser Gln Ile Ser Ser Ala Val Phe Leu
1795 1800 1805
Arg Arg Val Ala Leu Glu Val Ala Ser Pro Ala Gly Gln Gly Ala Leu
1810 1815 1820
Ala Ser Asn Val Glu Leu Cys Leu Cys Pro Ala Ser Tyr Arg Gly Asp
1825 1830 1835 1840
Ser Cys Gln Glu Cys Ala Pro Gly Phe Tyr Arg Asp Val Lys Gly Leu
1845 1850 1855
Phe Leu Gly Arg Cys Val Pro Cys Gln Cys His Gly His Ser Asp Arg
1860 1865 1870
Cys Leu Pro Gly Ser Gly Val Cys Val Asp Cys Gln His Asn Thr Glu
1875 1880 1885
Gly Ala His Cys Glu Arg Cys Gln Ala Gly Phe Val Ser Ser Arg Asp
1890 1895 1900
Asp Pro Ser Ala Pro Cys Val Ser Cys Pro Cys Pro Leu Ser Val Pro
1905 1910 1915 1920
Ser Asn Asn Phe Ala Glu Gly Cys Val Leu Arg Gly Gly Arg Thr Gln
1925 1930 1935
Cys Leu Cys Lys Pro Gly Tyr Ala Gly Ala Ser Cys Glu Arg Cys Ala
1940 1945 1950
Pro Gly Phe Phe Gly Asn Pro Leu Val Leu Gly Ser Ser Cys Gln Pro
1955 1960 1965
Cys Asp Cys Ser Gly Asn Gly Asp Pro Asn Leu Leu Phe Ser Asp Cys
1970 1975 1980
Asp Pro Leu Thr Gly Ala Cys Arg Gly Cys Leu Arg His Thr Thr Gly
1985 1990 1995 2000
Pro Arg Cys Glu Ile Cys Ala Pro Gly Phe Tyr Gly Asn Ala Leu Leu
2005 2010 2015
Pro Gly Asn Cys Thr Arg Cys Asp Cys Thr Pro Cys Gly Thr Glu Ala
2020 2025 2030
Cys Asp Pro His Ser Gly His Cys Leu Cys Lys Ala Gly Val Thr Gly
2035 2040 2045
Arg Arg Cys Asp Arg Cys Gln Glu Gly His Phe Gly Phe Asp Gly Cys
2050 2055 2060
Gly Gly Cys Arg Pro Cys Ala Cys Gly Pro Ala Ala Glu Gly Ser Glu
2065 2070 2075 2080
Cys His Pro Gln Ser Gly Gln Cys His Cys Arg Pro Gly Thr Met Gly
2085 2090 2095
Pro Gln Cys Arg Glu Cys Ala Pro Gly Tyr Trp Gly Leu Pro Glu Gln
2100 2105 2110
Gly Cys Arg Arg Cys Gln Cys Pro Gly Gly Arg Cys Asp Pro His Thr
2115 2120 2125
Gly Arg Cys Asn Cys Pro Pro Gly Leu Ser Gly Glu Arg Cys Asp Thr
2130 2135 2140
Cys Ser Gln Gln His Gln Val Pro Val Pro Gly Gly Pro Val Gly His
2145 2150 2155 2160
Ser Ile His Cys Glu Val Cys Asp His Cys Val Val Leu Leu Leu Asp
2165 2170 2175
Asp Leu Glu Arg Ala Gly Ala Leu Leu Pro Ala Ile His Glu Gln Leu
2180 2185 2190
Arg Gly Ile Asn Ala Ser Ser Met Ala Trp Ala Arg Leu His Arg Leu
2195 2200 2205
Asn Ala Ser Ile Ala Asp Leu Gln Ser Gln Leu Arg Ser Pro Leu Gly
2210 2215 2220
Pro Arg His Glu Thr Ala Gln Gln Leu Glu Val Leu Glu Gln Gln Ser
2225 2230 2235 2240
Thr Ser Leu Gly Gln Asp Ala Arg Arg Leu Gly Gly Gln Ala Val Gly
2245 2250 2255
Thr Arg Asp Gln Ala Ser Gln Leu Leu Ala Gly Thr Glu Ala Thr Leu
2260 2265 2270
Gly His Ala Lys Thr Leu Leu Ala Ala Ile Arg Ala Val Asp Arg Thr
2275 2280 2285
Leu Ser Glu Leu Met Ser Gln Thr Gly His Leu Gly Leu Ala Asn Ala
2290 2295 2300
Ser Ala Pro Ser Gly Glu Gln Leu Leu Arg Thr Leu Ala Glu Val Glu
2305 2310 2315 2320
Arg Leu Leu Trp Glu Met Arg Ala Arg Asp Leu Gly Ala Pro Gln Ala
2325 2330 2335
Ala Ala Glu Ala Glu Leu Ala Ala Ala Gln Arg Leu Leu Ala Arg Val
2340 2345 2350
Gln Glu Gln Leu Ser Ser Leu Trp Glu Glu Asn Gln Ala Leu Ala Thr
2355 2360 2365
Gln Thr Arg Asp Arg Leu Ala Gln His Glu Ala Gly Leu Met Asp Leu
2370 2375 2380
Arg Glu Ala Leu Asn Arg Ala Val Asp Ala Thr Arg Glu Ala Gln Glu
2385 2390 2395 2400
Leu Asn Ser Arg Asn Gln Glu Arg Leu Glu Glu Ala Leu Gln Arg Lys
2405 2410 2415
Gln Glu Leu Ser Arg Asp Asn Ala Thr Leu Gln Ala Thr Leu His Ala
2420 2425 2430
Ala Arg Asp Thr Leu Ala Ser Val Phe Arg Leu Leu His Ser Leu Asp
2435 2440 2445
Gln Ala Lys Glu Glu Leu Glu Arg Leu Ala Ala Ser Leu Asp Gly Ala
2450 2455 2460
Arg Thr Pro Leu Leu Gln Arg Met Gln Thr Phe Ser Pro Ala Gly Ser
2465 2470 2475 2480
Lys Leu Arg Leu Val Glu Ala Ala Glu Ala His Ala Gln Gln Leu Gly
2485 2490 2495
Gln Leu Ala Leu Asn Leu Ser Ser Ile Ile Leu Asp Val Asn Gln Asp
2500 2505 2510
Arg Leu Thr Gln Arg Ala Ile Glu Ala Ser Asn Ala Tyr Ser Arg Ile
2515 2520 2525
Leu Gln Ala Val Gln Ala Ala Glu Asp Ala Ala Gly Gln Ala Leu Gln
2530 2535 2540
Gln Ala Asp His Thr Trp Ala Thr Val Val Arg Gln Gly Leu Val Asp
2545 2550 2555 2560
Arg Ala Gln Gln Leu Leu Ala Asn Ser Thr Ala Leu Glu Glu Ala Met
2565 2570 2575
Leu Gln Glu Gln Gln Arg Leu Gly Leu Val Trp Ala Ala Leu Gln Gly
2580 2585 2590
Ala Arg Thr Gln Leu Arg Asp Val Arg Ala Lys Lys Asp Gln Leu Glu
2595 2600 2605
Ala His Ile Gln Ala Ala Gln Ala Met Leu Ala Met Asp Thr Asp Glu
2610 2615 2620
Thr Ser Lys Lys Ile Ala His Ala Lys Ala Val Ala Ala Glu Ala Gln
2625 2630 2635 2640
Asp Thr Ala Thr Arg Val Gln Ser Gln Leu Gln Ala Met Gln Glu Asn
2645 2650 2655
Val Glu Arg Trp Gln Gly Gln Tyr Glu Gly Leu Arg Gly Gln Asp Leu
2660 2665 2670
Gly Gln Ala Val Leu Asp Ala Gly His Ser Val Ser Thr Leu Glu Lys
2675 2680 2685
Thr Leu Pro Gln Leu Leu Ala Lys Leu Ser Ile Leu Glu Asn Arg Gly
2690 2695 2700
Val His Asn Ala Ser Leu Ala Leu Ser Ala Ser Ile Gly Arg Val Arg
2705 2710 2715 2720
Glu Leu Ile Ala Gln Ala Arg Gly Ala Ala Ser Lys Val Lys Val Pro
2725 2730 2735
Met Lys Phe Asn Gly Arg Ser Gly Val Gln Leu Arg Thr Pro Arg Asp
2740 2745 2750
Leu Ala Asp Leu Ala Ala Tyr Thr Ala Leu Lys Phe Tyr Leu Gln Gly
2755 2760 2765
Pro Glu Pro Glu Pro Gly Gln Gly Thr Glu Asp Arg Phe Val Met Tyr
2770 2775 2780
Met Gly Ser Arg Gln Ala Thr Gly Asp Tyr Met Gly Val Ser Leu Arg
2785 2790 2795 2800
Asp Lys Lys Val His Trp Val Tyr Gln Leu Gly Glu Ala Gly Pro Ala
2805 2810 2815
Val Leu Ser Ile Asp Glu Asp Ile Gly Glu Gln Phe Ala Ala Val Ser
2820 2825 2830
Leu Asp Arg Thr Leu Gln Phe Gly His Met Ser Val Thr Val Glu Arg
2835 2840 2845
Gln Met Ile Gln Glu Thr Lys Gly Asp Thr Val Ala Pro Gly Ala Glu
2850 2855 2860
Gly Leu Leu Asn Leu Arg Pro Asp Asp Phe Val Phe Tyr Val Gly Gly
2865 2870 2875 2880
Tyr Pro Ser Thr Phe Thr Pro Pro Pro Leu Leu Arg Phe Pro Gly Tyr
2885 2890 2895
Arg Gly Cys Ile Glu Met Asp Thr Leu Asn Glu Glu Val Val Ser Leu
2900 2905 2910
Tyr Asn Phe Glu Arg Thr Phe Gln Leu Asp Thr Ala Val Asp Arg Pro
2915 2920 2925
Cys Ala Arg Ser Lys Ser Thr Gly Asp Pro Trp Leu Thr Asp Gly Ser
2930 2935 2940
Tyr Leu Asp Gly Thr Gly Phe Ala Arg Ile Ser Phe Asp Ser Gln Ile
2945 2950 2955 2960
Ser Thr Thr Lys Arg Phe Glu Gln Glu Leu Arg Leu Val Ser Tyr Ser
2965 2970 2975
Gly Val Leu Phe Phe Leu Lys Gln Gln Ser Gln Phe Leu Cys Leu Ala
2980 2985 2990
Val Gln Glu Gly Ser Leu Val Leu Leu Tyr Asp Phe Gly Ala Gly Leu
2995 3000 3005
Lys Lys Ala Val Pro Leu Gln Pro Pro Pro Pro Leu Thr Ser Ala Ser
3010 3015 3020
Lys Ala Ile Gln Val Phe Leu Leu Gly Gly Ser Arg Lys Arg Val Leu
3025 3030 3035 3040
Val Arg Val Glu Arg Ala Thr Val Tyr Ser Val Glu Gln Asp Asn Asp
3045 3050 3055
Leu Glu Leu Ala Asp Ala Tyr Tyr Leu Gly Gly Val Pro Pro Asp Gln
3060 3065 3070
Leu Pro Pro Ser Leu Arg Arg Leu Phe Pro Thr Gly Gly Ser Val Arg
3075 3080 3085
Gly Cys Val Lys Gly Ile Lys Ala Leu Gly Lys Tyr Val Asp Leu Lys
3090 3095 3100
Arg Leu Asn Thr Thr Gly Val Ser Ala Gly Cys Thr Ala Asp Leu Leu
3105 3110 3115 3120
Val Gly Arg Ala Met Thr Phe His Gly His Gly Phe Leu Arg Leu Ala
3125 3130 3135
Leu Ser Asn Val Ala Pro Leu Thr Gly Asn Val Tyr Ser Gly Phe Gly
3140 3145 3150
Phe His Ser Ala Gln Asp Ser Ala Leu Leu Tyr Tyr Arg Ala Ser Pro
3155 3160 3165
Asp Gly Leu Cys Gln Val Ser Leu Gln Gln Gly Arg Val Ser Leu Gln
3170 3175 3180
Leu Leu Arg Thr Glu Val Lys Thr Gln Ala Gly Phe Ala Asp Gly Ala
3185 3190 3195 3200
Pro His Tyr Val Ala Phe Tyr Ser Asn Ala Thr Gly Val Trp Leu Tyr
3205 3210 3215
Val Asp Asp Gln Leu Gln Gln Met Lys Pro His Arg Gly Pro Pro Pro
3220 3225 3230
Glu Leu Gln Pro Gln Pro Glu Gly Pro Pro Arg Leu Leu Leu Gly Gly
3235 3240 3245
Leu Pro Glu Ser Gly Thr Ile Tyr Asn Phe Ser Gly Cys Ile Ser Asn
3250 3255 3260
Val Phe Val Gln Arg Leu Leu Gly Pro Gln Arg Val Phe Asp Leu Gln
3265 3270 3275 3280
Gln Asn Leu Gly Ser Val Asn Val Ser Thr Gly Cys Ala Pro Ala Leu
3285 3290 3295
Gln Ala Gln Thr Pro Gly Leu Gly Pro Arg Gly Leu Gln Ala Thr Ala
3300 3305 3310
Arg Lys Ala Ser Arg Arg Ser Arg Gln Pro Ala Arg His Pro Ala Cys
3315 3320 3325
Met Leu Pro Pro His Leu Arg Thr Thr Arg Asp Ser Tyr Gln Phe Gly
3330 3335 3340
Gly Ser Leu Ser Ser His Leu Glu Phe Val Gly Ile Leu Ala Arg His
3345 3350 3355 3360
Arg Asn Trp Pro Ser Leu Ser Met His Val Leu Pro Arg Ser Ser Arg
3365 3370 3375
Gly Leu Leu Leu Phe Thr Ala Arg Leu Arg Pro Gly Ser Pro Ser Leu
3380 3385 3390
Ala Leu Phe Leu Ser Asn Gly His Phe Val Ala Gln Met Glu Gly Leu
3395 3400 3405
Gly Thr Arg Leu Arg Ala Gln Ser Arg Gln Arg Ser Arg Pro Gly Arg
3410 3415 3420
Trp His Lys Val Ser Val Arg Trp Glu Lys Asn Arg Ile Leu Leu Val
3425 3430 3435 3440
Thr Asp Gly Ala Arg Ala Trp Ser Gln Glu Gly Pro His Arg Gln His
3445 3450 3455
Gln Gly Ala Glu His Pro Gln Pro His Thr Leu Phe Val Gly Gly Leu
3460 3465 3470
Pro Ala Ser Ser His Ser Ser Lys Leu Pro Val Thr Val Gly Phe Ser
3475 3480 3485
Gly Cys Val Lys Arg Leu Arg Leu His Gly Arg Pro Leu Gly Ala Pro
3490 3495 3500
Thr Arg Met Ala Gly Val Thr Pro Cys Ile Leu Gly Pro Leu Glu Ala
3505 3510 3515 3520
Gly Leu Phe Phe Pro Gly Ser Gly Gly Val Ile Thr Leu Asp Leu Pro
3525 3530 3535
Gly Ala Thr Leu Pro Asp Val Gly Leu Glu Leu Glu Val Arg Pro Leu
3540 3545 3550
Ala Val Thr Gly Leu Ile Phe His Leu Gly Gln Ala Arg Thr Pro Pro
3555 3560 3565
Tyr Leu Gln Leu Gln Val Thr Glu Lys Gln Val Leu Leu Arg Ala Asp
3570 3575 3580
Asp Gly Ala Gly Glu Phe Ser Thr Ser Val Thr Arg Pro Ser Val Leu
3585 3590 3595 3600
Cys Asp Gly Gln Trp His Arg Leu Ala Val Met Lys Ser Gly Asn Val
3605 3610 3615
Leu Arg Leu Glu Val Asp Ala Gln Ser Asn His Thr Val Gly Pro Leu
3620 3625 3630
Leu Ala Ala Ala Ala Gly Ala Pro Ala Pro Leu Tyr Leu Gly Gly Leu
3635 3640 3645
Pro Glu Pro Met Ala Val Gln Pro Trp Pro Pro Ala Tyr Cys Gly Cys
3650 3655 3660
Met Arg Arg Leu Ala Val Asn Arg Ser Pro Val Ala Met Thr Arg Ser
3665 3670 3675 3680
Val Glu Val His Gly Ala Val Gly Ala Ser Gly Cys Pro Ala Ala
3685 3690 3695
<210> 128
<211> 1786
<212> PRT
<213> 人工序列
<220>
<223> 层粘连蛋白亚单元beta-1的氨基酸序列(人)
<400> 128
Met Gly Leu Leu Gln Leu Leu Ala Phe Ser Phe Leu Ala Leu Cys Arg
1 5 10 15
Ala Arg Val Arg Ala Gln Glu Pro Glu Phe Ser Tyr Gly Cys Ala Glu
20 25 30
Gly Ser Cys Tyr Pro Ala Thr Gly Asp Leu Leu Ile Gly Arg Ala Gln
35 40 45
Lys Leu Ser Val Thr Ser Thr Cys Gly Leu His Lys Pro Glu Pro Tyr
50 55 60
Cys Ile Val Ser His Leu Gln Glu Asp Lys Lys Cys Phe Ile Cys Asn
65 70 75 80
Ser Gln Asp Pro Tyr His Glu Thr Leu Asn Pro Asp Ser His Leu Ile
85 90 95
Glu Asn Val Val Thr Thr Phe Ala Pro Asn Arg Leu Lys Ile Trp Trp
100 105 110
Gln Ser Glu Asn Gly Val Glu Asn Val Thr Ile Gln Leu Asp Leu Glu
115 120 125
Ala Glu Phe His Phe Thr His Leu Ile Met Thr Phe Lys Thr Phe Arg
130 135 140
Pro Ala Ala Met Leu Ile Glu Arg Ser Ser Asp Phe Gly Lys Thr Trp
145 150 155 160
Gly Val Tyr Arg Tyr Phe Ala Tyr Asp Cys Glu Ala Ser Phe Pro Gly
165 170 175
Ile Ser Thr Gly Pro Met Lys Lys Val Asp Asp Ile Ile Cys Asp Ser
180 185 190
Arg Tyr Ser Asp Ile Glu Pro Ser Thr Glu Gly Glu Val Ile Phe Arg
195 200 205
Ala Leu Asp Pro Ala Phe Lys Ile Glu Asp Pro Tyr Ser Pro Arg Ile
210 215 220
Gln Asn Leu Leu Lys Ile Thr Asn Leu Arg Ile Lys Phe Val Lys Leu
225 230 235 240
His Thr Leu Gly Asp Asn Leu Leu Asp Ser Arg Met Glu Ile Arg Glu
245 250 255
Lys Tyr Tyr Tyr Ala Val Tyr Asp Met Val Val Arg Gly Asn Cys Phe
260 265 270
Cys Tyr Gly His Ala Ser Glu Cys Ala Pro Val Asp Gly Phe Asn Glu
275 280 285
Glu Val Glu Gly Met Val His Gly His Cys Met Cys Arg His Asn Thr
290 295 300
Lys Gly Leu Asn Cys Glu Leu Cys Met Asp Phe Tyr His Asp Leu Pro
305 310 315 320
Trp Arg Pro Ala Glu Gly Arg Asn Ser Asn Ala Cys Lys Lys Cys Asn
325 330 335
Cys Asn Glu His Ser Ile Ser Cys His Phe Asp Met Ala Val Tyr Leu
340 345 350
Ala Thr Gly Asn Val Ser Gly Gly Val Cys Asp Asp Cys Gln His Asn
355 360 365
Thr Met Gly Arg Asn Cys Glu Gln Cys Lys Pro Phe Tyr Tyr Gln His
370 375 380
Pro Glu Arg Asp Ile Arg Asp Pro Asn Phe Cys Glu Arg Cys Thr Cys
385 390 395 400
Asp Pro Ala Gly Ser Gln Asn Glu Gly Ile Cys Asp Ser Tyr Thr Asp
405 410 415
Phe Ser Thr Gly Leu Ile Ala Gly Gln Cys Arg Cys Lys Leu Asn Val
420 425 430
Glu Gly Glu His Cys Asp Val Cys Lys Glu Gly Phe Tyr Asp Leu Ser
435 440 445
Ser Glu Asp Pro Phe Gly Cys Lys Ser Cys Ala Cys Asn Pro Leu Gly
450 455 460
Thr Ile Pro Gly Gly Asn Pro Cys Asp Ser Glu Thr Gly His Cys Tyr
465 470 475 480
Cys Lys Arg Leu Val Thr Gly Gln His Cys Asp Gln Cys Leu Pro Glu
485 490 495
His Trp Gly Leu Ser Asn Asp Leu Asp Gly Cys Arg Pro Cys Asp Cys
500 505 510
Asp Leu Gly Gly Ala Leu Asn Asn Ser Cys Phe Ala Glu Ser Gly Gln
515 520 525
Cys Ser Cys Arg Pro His Met Ile Gly Arg Gln Cys Asn Glu Val Glu
530 535 540
Pro Gly Tyr Tyr Phe Ala Thr Leu Asp His Tyr Leu Tyr Glu Ala Glu
545 550 555 560
Glu Ala Asn Leu Gly Pro Gly Val Ser Ile Val Glu Arg Gln Tyr Ile
565 570 575
Gln Asp Arg Ile Pro Ser Trp Thr Gly Ala Gly Phe Val Arg Val Pro
580 585 590
Glu Gly Ala Tyr Leu Glu Phe Phe Ile Asp Asn Ile Pro Tyr Ser Met
595 600 605
Glu Tyr Asp Ile Leu Ile Arg Tyr Glu Pro Gln Leu Pro Asp His Trp
610 615 620
Glu Lys Ala Val Ile Thr Val Gln Arg Pro Gly Arg Ile Pro Thr Ser
625 630 635 640
Ser Arg Cys Gly Asn Thr Ile Pro Asp Asp Asp Asn Gln Val Val Ser
645 650 655
Leu Ser Pro Gly Ser Arg Tyr Val Val Leu Pro Arg Pro Val Cys Phe
660 665 670
Glu Lys Gly Thr Asn Tyr Thr Val Arg Leu Glu Leu Pro Gln Tyr Thr
675 680 685
Ser Ser Asp Ser Asp Val Glu Ser Pro Tyr Thr Leu Ile Asp Ser Leu
690 695 700
Val Leu Met Pro Tyr Cys Lys Ser Leu Asp Ile Phe Thr Val Gly Gly
705 710 715 720
Ser Gly Asp Gly Val Val Thr Asn Ser Ala Trp Glu Thr Phe Gln Arg
725 730 735
Tyr Arg Cys Leu Glu Asn Ser Arg Ser Val Val Lys Thr Pro Met Thr
740 745 750
Asp Val Cys Arg Asn Ile Ile Phe Ser Ile Ser Ala Leu Leu His Gln
755 760 765
Thr Gly Leu Ala Cys Glu Cys Asp Pro Gln Gly Ser Leu Ser Ser Val
770 775 780
Cys Asp Pro Asn Gly Gly Gln Cys Gln Cys Arg Pro Asn Val Val Gly
785 790 795 800
Arg Thr Cys Asn Arg Cys Ala Pro Gly Thr Phe Gly Phe Gly Pro Ser
805 810 815
Gly Cys Lys Pro Cys Glu Cys His Leu Gln Gly Ser Val Asn Ala Phe
820 825 830
Cys Asn Pro Val Thr Gly Gln Cys His Cys Phe Gln Gly Val Tyr Ala
835 840 845
Arg Gln Cys Asp Arg Cys Leu Pro Gly His Trp Gly Phe Pro Ser Cys
850 855 860
Gln Pro Cys Gln Cys Asn Gly His Ala Asp Asp Cys Asp Pro Val Thr
865 870 875 880
Gly Glu Cys Leu Asn Cys Gln Asp Tyr Thr Met Gly His Asn Cys Glu
885 890 895
Arg Cys Leu Ala Gly Tyr Tyr Gly Asp Pro Ile Ile Gly Ser Gly Asp
900 905 910
His Cys Arg Pro Cys Pro Cys Pro Asp Gly Pro Asp Ser Gly Arg Gln
915 920 925
Phe Ala Arg Ser Cys Tyr Gln Asp Pro Val Thr Leu Gln Leu Ala Cys
930 935 940
Val Cys Asp Pro Gly Tyr Ile Gly Ser Arg Cys Asp Asp Cys Ala Ser
945 950 955 960
Gly Tyr Phe Gly Asn Pro Ser Glu Val Gly Gly Ser Cys Gln Pro Cys
965 970 975
Gln Cys His Asn Asn Ile Asp Thr Thr Asp Pro Glu Ala Cys Asp Lys
980 985 990
Glu Thr Gly Arg Cys Leu Lys Cys Leu Tyr His Thr Glu Gly Glu His
995 1000 1005
Cys Gln Phe Cys Arg Phe Gly Tyr Tyr Gly Asp Ala Leu Gln Gln Asp
1010 1015 1020
Cys Arg Lys Cys Val Cys Asn Tyr Leu Gly Thr Val Gln Glu His Cys
1025 1030 1035 1040
Asn Gly Ser Asp Cys Gln Cys Asp Lys Ala Thr Gly Gln Cys Leu Cys
1045 1050 1055
Leu Pro Asn Val Ile Gly Gln Asn Cys Asp Arg Cys Ala Pro Asn Thr
1060 1065 1070
Trp Gln Leu Ala Ser Gly Thr Gly Cys Asp Pro Cys Asn Cys Asn Ala
1075 1080 1085
Ala His Ser Phe Gly Pro Ser Cys Asn Glu Phe Thr Gly Gln Cys Gln
1090 1095 1100
Cys Met Pro Gly Phe Gly Gly Arg Thr Cys Ser Glu Cys Gln Glu Leu
1105 1110 1115 1120
Phe Trp Gly Asp Pro Asp Val Glu Cys Arg Ala Cys Asp Cys Asp Pro
1125 1130 1135
Arg Gly Ile Glu Thr Pro Gln Cys Asp Gln Ser Thr Gly Gln Cys Val
1140 1145 1150
Cys Val Glu Gly Val Glu Gly Pro Arg Cys Asp Lys Cys Thr Arg Gly
1155 1160 1165
Tyr Ser Gly Val Phe Pro Asp Cys Thr Pro Cys His Gln Cys Phe Ala
1170 1175 1180
Leu Trp Asp Val Ile Ile Ala Glu Leu Thr Asn Arg Thr His Arg Phe
1185 1190 1195 1200
Leu Glu Lys Ala Lys Ala Leu Lys Ile Ser Gly Val Ile Gly Pro Tyr
1205 1210 1215
Arg Glu Thr Val Asp Ser Val Glu Arg Lys Val Ser Glu Ile Lys Asp
1220 1225 1230
Ile Leu Ala Gln Ser Pro Ala Ala Glu Pro Leu Lys Asn Ile Gly Asn
1235 1240 1245
Leu Phe Glu Glu Ala Glu Lys Leu Ile Lys Asp Val Thr Glu Met Met
1250 1255 1260
Ala Gln Val Glu Val Lys Leu Ser Asp Thr Thr Ser Gln Ser Asn Ser
1265 1270 1275 1280
Thr Ala Lys Glu Leu Asp Ser Leu Gln Thr Glu Ala Glu Ser Leu Asp
1285 1290 1295
Asn Thr Val Lys Glu Leu Ala Glu Gln Leu Glu Phe Ile Lys Asn Ser
1300 1305 1310
Asp Ile Arg Gly Ala Leu Asp Ser Ile Thr Lys Tyr Phe Gln Met Ser
1315 1320 1325
Leu Glu Ala Glu Glu Arg Val Asn Ala Ser Thr Thr Glu Pro Asn Ser
1330 1335 1340
Thr Val Glu Gln Ser Ala Leu Met Arg Asp Arg Val Glu Asp Val Met
1345 1350 1355 1360
Met Glu Arg Glu Ser Gln Phe Lys Glu Lys Gln Glu Glu Gln Ala Arg
1365 1370 1375
Leu Leu Asp Glu Leu Ala Gly Lys Leu Gln Ser Leu Asp Leu Ser Ala
1380 1385 1390
Ala Ala Glu Met Thr Cys Gly Thr Pro Pro Gly Ala Ser Cys Ser Glu
1395 1400 1405
Thr Glu Cys Gly Gly Pro Asn Cys Arg Thr Asp Glu Gly Glu Arg Lys
1410 1415 1420
Cys Gly Gly Pro Gly Cys Gly Gly Leu Val Thr Val Ala His Asn Ala
1425 1430 1435 1440
Trp Gln Lys Ala Met Asp Leu Asp Gln Asp Val Leu Ser Ala Leu Ala
1445 1450 1455
Glu Val Glu Gln Leu Ser Lys Met Val Ser Glu Ala Lys Leu Arg Ala
1460 1465 1470
Asp Glu Ala Lys Gln Ser Ala Glu Asp Ile Leu Leu Lys Thr Asn Ala
1475 1480 1485
Thr Lys Glu Lys Met Asp Lys Ser Asn Glu Glu Leu Arg Asn Leu Ile
1490 1495 1500
Lys Gln Ile Arg Asn Phe Leu Thr Gln Asp Ser Ala Asp Leu Asp Ser
1505 1510 1515 1520
Ile Glu Ala Val Ala Asn Glu Val Leu Lys Met Glu Met Pro Ser Thr
1525 1530 1535
Pro Gln Gln Leu Gln Asn Leu Thr Glu Asp Ile Arg Glu Arg Val Glu
1540 1545 1550
Ser Leu Ser Gln Val Glu Val Ile Leu Gln His Ser Ala Ala Asp Ile
1555 1560 1565
Ala Arg Ala Glu Met Leu Leu Glu Glu Ala Lys Arg Ala Ser Lys Ser
1570 1575 1580
Ala Thr Asp Val Lys Val Thr Ala Asp Met Val Lys Glu Ala Leu Glu
1585 1590 1595 1600
Glu Ala Glu Lys Ala Gln Val Ala Ala Glu Lys Ala Ile Lys Gln Ala
1605 1610 1615
Asp Glu Asp Ile Gln Gly Thr Gln Asn Leu Leu Thr Ser Ile Glu Ser
1620 1625 1630
Glu Thr Ala Ala Ser Glu Glu Thr Leu Phe Asn Ala Ser Gln Arg Ile
1635 1640 1645
Ser Glu Leu Glu Arg Asn Val Glu Glu Leu Lys Arg Lys Ala Ala Gln
1650 1655 1660
Asn Ser Gly Glu Ala Glu Tyr Ile Glu Lys Val Val Tyr Thr Val Lys
1665 1670 1675 1680
Gln Ser Ala Glu Asp Val Lys Lys Thr Leu Asp Gly Glu Leu Asp Glu
1685 1690 1695
Lys Tyr Lys Lys Val Glu Asn Leu Ile Ala Lys Lys Thr Glu Glu Ser
1700 1705 1710
Ala Asp Ala Arg Arg Lys Ala Glu Met Leu Gln Asn Glu Ala Lys Thr
1715 1720 1725
Leu Leu Ala Gln Ala Asn Ser Lys Leu Gln Leu Leu Lys Asp Leu Glu
1730 1735 1740
Arg Lys Tyr Glu Asp Asn Gln Arg Tyr Leu Glu Asp Lys Ala Gln Glu
1745 1750 1755 1760
Leu Ala Arg Leu Glu Gly Glu Val Arg Ser Leu Leu Lys Asp Ile Ser
1765 1770 1775
Gln Lys Val Ala Val Tyr Ser Thr Cys Leu
1780 1785
<210> 129
<211> 34
<212> DNA
<213> 人工序列
<220>
<223> scFv VH 正向引物
<400> 129
agagagtgta cactcccagg cggccgaggt gcag 34
<210> 130
<211> 42
<212> DNA
<213> 人工序列
<220>
<223> scFv VH 反向引物
<400> 130
cgccgctggg cccttggtgg aggctgagct cacggtgacc ag 42
<210> 131
<211> 89
<212> DNA
<213> 人工序列
<220>
<223> scFv VL 正向引物
<400> 131
aagcggccgc caccatggga tggagctgta tcatcctctt cttggtagca acagctacag 60
gtgtacactc ccagtctgtg ctgactcag 89
<210> 132
<211> 30
<212> DNA
<213> 人工序列
<220>
<223> scFv VL 反向引物
<400> 132
cgccgccgta cgtaggaccg tcagcttggt 30
<210> 133
<211> 29
<212> PRT
<213> 人工序列
<220>
<223> N3 表位 F1
<400> 133
Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Ser Glu Pro
1 5 10 15
Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala
20 25
<210> 134
<211> 30
<212> PRT
<213> 人工序列
<220>
<223> N3 表位 F2
<400> 134
Gln Ala Ala Glu Val Lys Val Asp Gly Ser Glu Pro Lys Leu Ser Lys
1 5 10 15
Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Val Ala
20 25 30
<210> 135
<211> 29
<212> PRT
<213> 人工序列
<220>
<223> N3 表位 F3
<400> 135
Lys Val Asp Gly Ser Glu Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg
1 5 10 15
Arg Leu Lys Ala Glu Lys Lys Val Ala Glu Lys Glu Ala
20 25
<210> 136
<211> 28
<212> PRT
<213> 人工序列
<220>
<223> N3 表位 F4
<400> 136
Glu Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu
1 5 10 15
Lys Lys Val Ala Glu Lys Glu Ala Lys Gln Lys Glu
20 25
<210> 137
<211> 26
<212> PRT
<213> 人工序列
<220>
<223> N3 表位 F5
<400> 137
Lys Arg Arg Leu Lys Ala Glu Lys Lys Val Ala Glu Lys Glu Ala Lys
1 5 10 15
Gln Lys Glu Leu Ser Glu Lys Gln Leu Ser
20 25
<210> 138
<211> 28
<212> PRT
<213> 人工序列
<220>
<223> 小鼠 N3 表位 F1(mF1)
<400> 138
Met Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly Glu Gln Lys
1 5 10 15
Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala
20 25
<210> 139
<211> 29
<212> PRT
<213> 人工序列
<220>
<223> 小鼠 N3 表位 F2(mF2)
<400> 139
Gln Glu Ser Glu Val Lys Val Asp Gly Glu Gln Lys Leu Ser Lys Asn
1 5 10 15
Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Leu Ala
20 25
<210> 140
<211> 28
<212> PRT
<213> 人工序列
<220>
<223> 小鼠 N3 表位 F3(mF3)
<400> 140
Lys Val Asp Gly Glu Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg
1 5 10 15
Leu Lys Ala Glu Lys Lys Leu Ala Glu Lys Glu Ala
20 25
<210> 141
<211> 27
<212> PRT
<213> 人工序列
<220>
<223> 小鼠 N3 表位 F4(mF4)
<400> 141
Gln Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys
1 5 10 15
Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu
20 25
<210> 142
<211> 25
<212> PRT
<213> 人工序列
<220>
<223> 小鼠 N3 表位 F5(mF5)
<400> 142
Arg Arg Leu Lys Ala Glu Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln
1 5 10 15
Lys Glu Leu Ser Glu Lys Gln Leu Asn
20 25
<210> 143
<211> 28
<212> PRT
<213> 人工序列
<220>
<223> 大鼠 N3 表位 F1(rF1)
<400> 143
Met Ala Thr Leu Arg Glu Gly Glu Val Lys Leu Asp Gly Glu Pro Lys
1 5 10 15
Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala
20 25
<210> 144
<211> 29
<212> PRT
<213> 人工序列
<220>
<223> 大鼠 N3 表位 F2(rF2)
<400> 144
Arg Glu Gly Glu Val Lys Leu Asp Gly Glu Pro Lys Leu Ser Lys Asn
1 5 10 15
Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Leu Ala
20 25
<210> 145
<211> 28
<212> PRT
<213> 人工序列
<220>
<223> 大鼠 N3 表位 F3(rF3)
<400> 145
Lys Leu Asp Gly Glu Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg
1 5 10 15
Leu Lys Ala Glu Lys Lys Leu Ala Glu Lys Glu Ala
20 25
<210> 146
<211> 27
<212> PRT
<213> 人工序列
<220>
<223> 大鼠 N3 表位 F4(rF4)
<400> 146
Pro Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys
1 5 10 15
Lys Leu Ala Glu Lys Glu Ala Lys Gln Lys Glu
20 25
<210> 147
<211> 25
<212> PRT
<213> 人工序列
<220>
<223> 大鼠 N3 表位 F5(rF5)
<400> 147
Arg Arg Leu Lys Ala Glu Lys Lys Leu Ala Glu Lys Glu Ala Lys Gln
1 5 10 15
Lys Glu Leu Ser Glu Lys Gln Leu Asn
20 25
<210> 148
<211> 52
<212> PRT
<213> 人工序列
<220>
<223> 暴露于膜的赖氨酰-tRNA合成酶N-末端(人)
<400> 148
Met Ala Ala Val Gln Ala Ala Glu Val Lys Val Asp Gly Ser Glu Pro
1 5 10 15
Lys Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys
20 25 30
Val Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu
35 40 45
Ser Gln Ala Thr
50
<210> 149
<211> 48
<212> PRT
<213> 人工序列
<220>
<223> 赖氨酰-tRNA合成酶N-末端(小鼠)
<400> 149
Met Ala Thr Leu Gln Glu Ser Glu Val Lys Val Asp Gly Glu Gln Lys
1 5 10 15
Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Leu
20 25 30
Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu Asn
35 40 45
<210> 150
<211> 48
<212> PRT
<213> 人工序列
<220>
<223> 赖氨酰-tRNA合成酶N-末端(大鼠)
<400> 150
Met Ala Thr Leu Arg Glu Gly Glu Val Lys Leu Asp Gly Glu Pro Lys
1 5 10 15
Leu Ser Lys Asn Glu Leu Lys Arg Arg Leu Lys Ala Glu Lys Lys Leu
20 25 30
Ala Glu Lys Glu Ala Lys Gln Lys Glu Leu Ser Glu Lys Gln Leu Asn
35 40 45
<210> 151
<211> 17
<212> PRT
<213> 人工序列
<220>
<223> CDR H2 突变体 5 氨基酸
<400> 151
Val Ile Ser Ser Asp Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys
1 5 10 15
Gly
<210> 152
<211> 51
<212> DNA
<213> 人工序列
<220>
<223> CDR H2 突变体 5 DNA
<400> 152
gtgatctctt ctgatggtgg taatacatat tacgctgatt ctgtaaaagg t 51