CN1495195A - 表达雄激素受体复合物相关蛋白的转基因动物 - Google Patents
表达雄激素受体复合物相关蛋白的转基因动物 Download PDFInfo
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Abstract
本发明涉及表达雄激素受体复合物相关蛋白基因的转基因动物。这些动物可以作为开发治疗癌症,例如肝癌的药物的动物模型。
Description
相关申请
本申请是2001年2月12日提交的美国专利申请09/781,693的专利申请和2001年1月17日提交的美国临时申请60/262,312的部分继续申请,因此该申请要求上述两项的优先权,它们的内容也一并纳入本文作参考。
背景技术
目前已鉴定出相对于健康细胞而言在肿瘤细胞中过表达的多种基因。人们期望这些基因的鉴定可为抗癌药物的开发和癌症的诊断提供药物标靶。在肝脏肿瘤细胞中,类固醇受体(例如雄激素受体)的数量显示高于它们邻近的健康的肝细胞。
类固醇激素通常通过与它们的特异性核受体结合形成复合物,并由该复合物作为转录因子而发挥生理作用。所述复合物结合类固醇应答基因启动子中的特定核苷酸序列(类固醇反应元件),从而促进这些基因的转录。
发明简述
本发明的是基于编码雄激素受体复合物相关蛋白(ARCAP)的小鼠基因的发现,该基因有85%与人ARCAP相同。已发现人ARCAP在肝细胞癌细胞中过表达(与邻近的正常细胞相比),并与雄激素受体结合,从而增强该受体反式激活(transactivate)雄激素应答基因的能力。作为人ARCAP基因的同系物,小鼠ARCAP基因能用来产生可用作开发癌症(例如肝癌)治疗药物的动物模型的转基因小鼠。
小鼠ARCAP的全长cDNA(称为SEQ ID NO:1)如下所示,其中的起始和终止密码子用下划线标示:
GAGAGTATGAGGCGAGCCGTGGCCCGGGTGCCTCTCCGCTCCCGGGGAGA
AGGCGCGGCCGTGGCTGCCGCCCTCTGAGTCGCGGCGCCGGCGAGGCCCC
GGGGCGCGCGCATGGTGCTGGTGCCGCTCGGGTGTTGATCGGCCTGTCCC
CTCCCTCTCTTCCCCTCCCCACCCCCCGCGGTGGTCTCCCCTTTCCCACC
CCAGCCCTTGCGGAGCC
ATGGCTCGGAGTGGCTCCTGCCCGCACCTGTTG
TGGGACGTGAGGAAAAGGTCCCTTGGGCTGGAGGACCCGTCCCGGCTGAG
GAGCCGCTACCTGGGAAGAAGAGAATTTATCCAAAGATTAAAACTTGAAG
CAACTTTAAATGTGCATGATGGCTGTGTTAATACAATCTGTTGGAATGAC
ACTGGAGAATATATTTTATCTGGCTCTGATGACACTAAACTTGTAATTAG
TAATCCATACAGCAGAAAGGTTTTGACAACCATCCGTTCAGGGCATCGAG
CAAATATATTTAGTGCAAAGTTTTTGCCGTGCACAGATGATAAGCAGATT
GTGTCTTGCTCTGGAGATGGAGTCATATTTTATACTAACATTGAGCAAGA
TGCAGAAACTAACAGACAGTGCCAATTTACGTGCCATTATGGAACTACTT
ATGAGATTATGACTGTACCAAACGACCCTTACACCTTTCTGTCCTGTGGT
GAAGATGGAACTGTTAGGTGGTTTGACACACGCATCAAAACCAGTTGCAC
AAAAGAAGACTGTAAAGATGATATTTTAATCAACTGTAGGCGTGCTGCCA
CATCTGTGGCTATTTGTCCCCCAGTACCATATTACCTTGCTGTGGGTTGT
TCTGACAGCTCAGTACGGATTTATGATCGGCGAATGCTGGGCACAAGAGC
TACAGGGAATTATGCAGGCCGAGGAACTACTGGAATGGTTGCTCGATTTA
TACCTTCTCATCTTAGTAACAAATCATGCAGAGTGACATCACTGTGTTAC
AGTGAAGATGGTCAAGAGATTCTTGTCAGTTATTCTTCAGACTACATCTA
TCTTTTTGACCCCAAAGATGATACTGCACGAGAACTTAAAACTCCTTCTG
CAGAGGAGAGGCGAGAAGAGTTACGACAGCCTCCAGTTAAGCGCTTGAGA
CTTCGTGGTGATTGGTCAGATACTGGTCCCAGAGCACGGCCAGAAAGTGA
ACGAGAACGAGATGGAGAGCAAAGTCCCAATGTGTCACTGATGCAGAGAA
TGTCTGATATGTTATCAAGGTGGTTTGAAGAAGCAAGTGAAGTTGCACAA
AGCAACAGAGGAAGAGGAAGACCTCGGCCCAGAGGTGGAACAAATCAGCC
AGATGTTTCAACTCTTCCTACGGTTCCATCAAGTCCTAATTTGGAAGTGT
GTGAAACTGCAATGGATGTAGACATGCCAGCTGCACTTCTTCAGCCTTCT
ACATCCTCTACAGATCCAGTTCAGGCTCAGGCAGCCACAGCCGCCATAGA
AAGCCCTCGTTCCAGCTCGTTGCTGTCTTGCCCAGACAGTGAACCGAGGC
AGTCTGTTGAGGCGTCTGGACACCATGCACATCATCAGTCAGATTCTCCT
TCTTCTGTGGTTAACAAACAGCTGGGATCCATGTCACTTGATGAGCAACA
GGATAACAGTAATGAGAGGCTGAGCCCCAAACCAGGGACAGGTGAACCCG
TTTTAAGTTTGCACTACAGCACAGAAGGAACAACTACAAGCACAATAAAA
CTGAACTTTACAGATGAATGGAGCAGTACAGCCTCAAGTTCCAGAGGAAA
TGGGAGCCATTGCAAATCTGAGGGTCAGGAAGAATGCTTGGTCCCTCCGA
GCTCTGTGCAGCCACCGGAAGGAGACAGTGAAACAAGAGCTCCTGAAGAA
CTATCAGAGAAAGGAACACTTCCAGAAAACCTCACTCAAAACCAGATAGA
TACAGCACAACTTGATAACTTCCCAGCTGAGCCATTGGATTCTAACTCAG
GAGAGAAGAATAACCCAAGTCAGGACAGCCCTTGTGGGCTTCCAGAAGAA
GGCACTTTGTCTGAAACAGACAGGGAGACTTGTGAGCAGGCCAGCACTGA
GAGTGCTACCAGGCATGCTAGCACCAAGCCTGAACTCCCATCCCAGACAG
AAGCCATTGAGCAGGCCAGCACTGAGAGTGCTACCAGGCATACCAGTGCC
AATCCTGAACTCCCATCCCAGACAGAAGCCATAGCACCTTTAGCTCATGA
AGACCCATCTGCCAGGGACTCTGCTCTCCAGGACACAGATGACAGCGATG
ATGATCCGGTCTTGATCCCTGGTGCAAGATACCGAACAGGACCTGGTGAT
AGACGCTCCGCTGTTGCCCGCATTCAGGAGTTCTTCAGGAGGAGAAAAGA
AAGGAAAGAAATGGAAGAGCTGGATACTTTGAACATTAGGAGGCCACTAG
TAAAGATGGTTTATAAGGGCCACCGCAACTCCCGGACAATGATAAAAGAA
GCCAATTTCTGGGGTGCTAACTTTGTAATGAGCGGTTCCGATTGTGGCCA
TATCTTCATCTGGGACCGGCACACTGCGGAGCATTTGATGCTTCTGGAAG
CTGATAATCATGTGGTCAACTGCCTGCAGCCCCATCCGTTTGACCCAATT
CTAGCCTCATCTGGCATAGATTATGACATAAAGATCTGGTCGCCACTAGA
AGAGTCAAGAATTTTTAATCGAAAACTTGCTGATGAAGTTATAACTCGGA
ATGAACTCACGCTGGAAGAGACTCGGAACACCATCACCGTCCCAGCCTCT
TTCATGTTGAGGATGTTGGCGTCACTGAATCATATCCGAGCTGACCGTCT
GGAGGGTGACAGATCAGAAGGTTCAGGTCAGGAGAATGAAAATGAGGATG
AAGAA
TAAAGAACTCCTTGGCAAGCACTTAGATGTTCTGAGATTTGTATA
CGACATTTATTATATTTTTTTTCTTTACAGAACTTTAGTGCAATTTAAGG
CTATGGGTTTTTTTTTTTCTTTTTTTTTGGAGTTCTTCCCTATTTTGGGG
ATAACCAAACATTGGTTTGGAATGAGTGTGTGCATGAGTTGGGAGAGTGT
GTAAAACAAAGTAAGCAAAATGTTTTTTGAAACCTTTTGCCGTGTATGGA
GTCCCAAAAAAAAAAAAAAAAAAAAAAAAAAGCAAAGTGCAATACTTCCT
GACCCTCCGCTGTGGGAGCTTGGATCAATGCTGAAGTCATTTTCATTGTA
GTGAAAACGTTGGTTCAAATAAATTTCTACACTTGCCATTTGCAAAAAAA
AAAAAAAAAAAAAAAAAAAAAAGCGGCCGCTGAATTCTAG(SEQ ID NO:1)
编码小鼠ARCAP蛋白的核苷酸序列(即SEQ ID NO:1中从ATG起始密码子至终止密码子之前紧接的密码子)命名为SEQ ID NO:2。由上述cDNA编码的小鼠ARCAP蛋白(命名为SEQ ID NO:3)如下所示:
Met ala arg ser gly ser cys pro his leu leu trp asp val arg lys arg ser leu gly
leu glu asp pro ser arg leu arg ser arg tyr leu gly arg arg glu phe ile gln arg
leu lys leu glu ala thr leu asn val his asp gly cys val asn thr ile cys trp asn
asp thr gly glu tyr ile leu ser gly ser asp asp thr lys leu val ile ser asn pro
tyr ser arg lys val leu thr thr ile arg ser gly his arg ala asn ile phe ser ala
lys phe leu pro cys thr asp asp lys gln ile val ser cys ser gly asp gly val ile
phe tyr thr asn ile glu gln asp ala glu thr asn arg gln cys gln phe thr cys his
tyr gly thr thr tyr glu ile met thr val pro asn asp pro tyr thr phe leu ser cys
gly glu asp gly thr val arg trp phe asp thr arg ile lys thr ser cys thr lys glu
asp cys lys asp asp ile leu ile asn cys arg arg ala ala thr ser val ala ile cys
pro pro val pro tyr tyr leu ala val gly cys ser asp ser ser val arg ile tyr asp
arg arg met leu gly thr arg ala thr gly asn tyr ala gly arg gly thr thr gly met
val ala arg phe ile pro ser his leu ser asn lys ser cys arg val thr ser leu cys
tyr ser glu asp gly gln glu ile leu val ser tyr ser ser asp tyr ile tyr leu phe
asp pro lys asp asp thr ala arg glu leu lys thr pro ser ala glu glu arg arg glu
glu leu arg gln pro pro val lys arg leu arg leu arg gly asp trp ser asp thr gly
pro arg ala arg pro glu ser glu arg glu arg asp gly glu gln ser pro asn val ser
leu met gln arg met ser asp met leu ser arg trp phe glu glu ala ser glu val ala
gln ser asn arg gly arg gly arg pro arg pro arg gly gly thr asn gln pro asp val
ser thr leu pro thr val pro ser ser pro asn leu glu val cys glu thr ala met asp
val asp met pro ala ala leu leu gln pro ser thr ser ser thr asp pro val gln ala
gln ala ala thr ala ala ile glu ser pro arg ser ser ser leu leu ser cys pro asp
ser glu pro arg gln ser val glu ala ser gly his his ala his his gln ser asp ser
pro ser ser val val asn lys gln leu gly ser met ser leu asp glu gln gln asp asn
ser asn glu arg leu ser pro lys pro gly thr gly glu pro val leu ser leu his tyr
ser thr glu gly thr thr thr ser thr ile lys leu asn phe thr asp glu trp ser ser
thr ala ser ser ser arg gly asn gly ser his cys lys ser glu gly gln glu glu cys
leu val pro pro ser ser val gln pro pro glu gly asp ser glu thr arg ala pro glu
glu leu ser glu lys gly thr leu pro glu asn leu thr gln asn gln ile asp thr ala
gln leu asp asn phe pro ala glu pro leu asp ser asn ser gly glu lys asn asn pro
ser gln asp ser pro cys gly leu pro glu glu gly thr leu ser glu thr asp arg glu
thr cys glu gln ala ser thr glu ser ala thr arg his ala ser thr lys pro glu leu
pro ser gln thr glu ala ile glu gln ala ser thr glu ser ala thr arg his thr ser
ala asn pro glu leu pro ser gln thr glu ala ile ala pro leu ala his glu asp pro
ser ala arg asp ser ala leu gln asp thr asp asp ser asp asp asp pro val leu ile
pro gly ala arg tyr arg thr gly pro gly asp arg arg ser ala val ala arg ile gln
glu phe phe arg arg arg lys glu arg lys glu met glu glu leu asp thr leu asn ile
arg arg pro leu val lys met val tyr lys gly his arg asn ser arg thr met ile lys
glu ala asn phe trp gly ala asn phe val met ser gly ser asp cys gly his ile phe
ile trp asp arg his thr ala glu his leu met leu leu glu ala asp asn his val val
asn cys leu gln pro his pro phe asp pro ile leu ala ser ser gly ile asp tyr asp
ile lys ile trp ser pro leu glu glu ser arg ile phe asn arg lys leu ala asp glu
val ile thr arg asn glu leu thr leu glu glu thr arg asn thr ile thr val pro ala
ser phe met leu arg met leu ala ser leu asn his ile arg ala asp arg leu glu gly
asp arg ser glu gly ser gly gln glu asn glu asn glu asp glu glu(SEQID No:3)
因此,本发明的特征在于基本纯的多肽或蛋白,它们包括与SEQ IDNO:3至少70%(例如至少75,80,85,90,95,98或100%)相同的氨基酸序列。如果该多肽包括与SEQ ID NO:3 100%相同的序列,那么该多肽可包含多至30个保守氨基酸取代。本发明也包括基本纯的由在严谨条件下能与具有SEQ ID NO:2所示序列的探针杂交的核酸编码的多肽。所述多肽与雄激素受体结合,并增强雄激素受体反式激活雄激素应答基因的能力。这些多肽可用于制备ARCAP抗体(单克隆抗体或多克隆抗体)。这些抗体可用来检测组织或细胞区室中ARCAP的存在和分布。例如,这样的抗体可通过确定ARCAP蛋白在组织中是否表达或过表达来诊断癌性肝组织。它们也可如下所述用于治疗癌症(例如肝癌)
本发明的特征还在于分离的编码本发明多肽的核酸,包含本发明核酸的载体以及包含本发明核酸的细胞(在动物模型中或在培养物中)。本发明范围内的核酸的实例包括下述的分离的核酸,该核酸具有在严谨条件下与序列为SEQ ID NO:2或SEQ ID NO:2的互补体的单链探针杂交的链,该核酸的长度至少是15(例如至少是30,50,100,200,500或1000)个核苷酸。上述这些核酸,载体和细胞可用于制备动物模型,诊断癌症(例如肝癌),或制备本发明的多肽。例如,本发明的核酸可通过确定组织或细胞中ARCAPmRNA是否表达或过表达来诊断肝癌。该核酸还可用作基于PCR的检测方法中的探针,或在核酸印迹(例如Northern印迹)中用作带标记的探针。
本发明的特征还在于转基因动物(例如,啮齿类动物,如小鼠),其基因组包括含有本发明核酸的转基因,这种动物与野生型动物相比对雄激素的应答增强。该转基因动物可这样产生,即将本发明的核酸导入细胞,使得从该核酸产生转录物,并将该转录物翻译成蛋白。这些转基因动物可用作开发癌症(例如肝癌)治疗药物的模型。
此外,本发明的特征还在于制备本发明多肽的方法,即培养上述细胞,在该细胞中表达所述多肽,并从培养物中分离出这种多肽。
本发明的特征也在于检测动物样品(例如血样品)中是否包含癌细胞的方法。该方法涉及提供来自动物(例如,啮齿类动物,如小鼠)的样品,检测该样品中ARCAP基因的表达水平。如果该样品中ARCAP基因的表达水平高于正常样品中的水平,便表明该动物样品中含有癌细胞(例如肝癌细胞)。这种方法可用于诊断癌症或监控动物模型中癌症的治疗。
治疗动物(例如,啮齿类动物,如小鼠)的癌症(例如肝癌)的方法也包括在本发明的范围之内。所述方法涉及鉴定具有表达ARCAP基因的癌细胞的动物,并用可阻断ARCAP与雄激素受体的结合或降低雄激素受体反式激活雄激素应答基因的能力的组合物治疗所述动物。该组合物可包含本发明的抗体或本发明的反义核酸。这种方法也可用于筛选药物(包括用于基因治疗的药物)和测试它们在动物模型中的效力。
本文针对给定多肽所用的术语“基本纯的”是指该多肽基本不含有其他生物大分子。基本纯的多肽是干重为至少75%(例如至少80,85,95或99%)纯。纯度可通过任何适当的标准方法来检测,例如柱层析,聚丙烯酰胺凝胶电泳或HPLC分析。
“保守氨基酸取代”是指一种氨基酸残基被另一种具有化学相似侧链的氨基酸残基所取代。本领域已确定了具有类似侧链的氨基酸残基的家族。这些家族包括具有碱性侧链的氨基酸(例如,赖氨酸,精氨酸,组氨酸),具有酸性侧链的氨基酸(例如天冬氨酸,谷氨酸),具有不带电荷的极性侧链的氨基酸(例如甘氨酸,天冬酰胺,谷氨酰胺,丝氨酸,苏氨酸,酪氨酸,半胱氨酸),具有非极性侧链的氨基酸(例如丙氨酸,缬氨酸,亮氨酸,异亮氨酸,脯氨酸,苯丙氨酸,蛋氨酸,色氨酸),具有β分支侧链的氨基酸(例如苏氨酸,缬氨酸,异亮氨酸)以及具有芳香族侧链的氨基酸(例如酪氨酸,苯丙氨酸,色氨酸,组氨酸)。
在“严谨条件下”杂交是指在65℃,0.5×SSC杂交,然后在45℃,0.1×SSC洗涤。
两个氨基酸序列或核苷酸序列的“同一性百分比”利用Karlin和Altschul的算法(Proc.Natl.Acad.Sci.USA 87:2264-2268,1990),经Karlin和Altschul修改后的算法(Proc.Natl.Acad.Sci.USA 90:5873-5877,1993)确定。这种算法被纳入到Altschul等的NBLAST和XBLAST程序中(J.Mol.Biol.215:403-410,1990)。BLAST核苷酸检索用NBLAST程序进行,分值(score)=100,字段长(wordlength)=12。用XBLAST程序进行BLAST蛋白的检索,分值=50,字段长=3。两序列间存在缺口时,使用Altschul等所述的GappedBLAST(Nucleic Acids Res.25:3389-3402,1997)。当使用BLAST和GappedBLAST程序时,采用各自程序(例如XBLAST和NBLAST)的默认参数,参见
www.ncbi.nlm.nih.gov。
“分离的核酸”是这样一种核酸,其结构与任何天然核酸的结构不同,或与跨越3个以上被分隔的基因(separate gene)的天然基因组核酸的任何片段不同。因此,该术语包括,例如,(a)一种核酸,其具有天然基因组DNA分子的部分序列,但其侧翼并非天然生物基因组中该分子所述部分的侧翼编码序列;(b)掺入到载体或原核或真核生物基因组DNA中的核酸,其掺入方式致使所得分子与任何天然载体或基因组DNA都不同;(c)分离的分子如cDNA,基因组片段,由聚合酶链反应(PCR)制备的片段,或限制性片段;以及(d)重组核苷酸序列,其是杂合基因(即编码融合蛋白的基因)的一部分。特别排除在此定义之外的是存在于不同的(i)DNA分子,(ii)经转染的细胞或(iii)细胞克隆组成的混合物中的核酸,例如,DNA文库,如cDNA或基因组DNA文库中的核酸。
本发明的其他特征或优点可从以下详细说明以及权利要求中明显看出。
发明详述
本发明涉及表达ARCAP基因的非人转基因动物,以及利用所述动物开发治疗或预防肝癌的药物的方法。
本文所用的“非人转基因动物”包括起始的(founder)非人转基因动物,其后代以及来源于所述动物的细胞和组织。非人转基因动物可以是农场动物(farm animals),如猪,山羊,绵羊,牛,马和兔,啮齿类动物如大鼠,豚鼠和小鼠,以及非人灵长类动物如狒狒(baboon),猴子(monkey)和黑猩猩(chimpanzee)。转基因的猪和小鼠尤其有用。
本发明的非人转基因动物包含整合到其基因组中的编码ARCAP蛋白的核酸。本文所用的“ARCAP蛋白”是指野生型ARCAP蛋白或功能等价的变体,例如全长蛋白的片段。
本发明认为鉴定基因组中的内含子序列(如果有),并把它们包括在编码ARCAP蛋白的核酸中是有利的。
在本发明的非人转基因动物中,所述核酸可操作连接于促进ARCAP基因在,例如,肝脏中表达的调控元件。本文所用术语“可操作连接”是指调控元件和核酸所处的位置关系,它能使所述核酸被转录。所述调控元件可以是肝特异的启动子,如PEPCK和清蛋白启动子。
本发明的特征还在于适于产生本发明非人转基因动物的表达载体。所述表达载体包括能介导肝脏表达并与上述编码ARCAP蛋白的核酸可操作连接的启动子。
可用本领域已知的各种技术将表达载体导入到非人动物中,以产生非人转基因动物的起始动物。所述技术包括但不限于,原核显微注射(美国专利4,873,191),反转录病毒介导的基因向生殖系(germ line)中转移(Van derPutten et al.,Proc.Natl.Acad.Sci.USA,82:6148,1985),基因靶向进入胚胎干细胞(Thompson et al.,Cell,56:313,1989),胚胎电穿孔(Lo,Mol.Cell.Biol.,3:1803,1983),和体外体细胞转化及后续的核移植(Wilmut et al.,Nature,385(6619):810-813,1997;and Wakayama et al.,Nature,394:369-374,1998)。在一个实施例中,表达载体被显微注射到非人动物的卵或胚胎中,或显微注射到非人动物的胚胎干细胞中。
一旦制备出非人转基因动物,(例如依照Current Protocol(Wiley,USA)和Manuplate the Mouse Embryo(Hogan Beddington and Costantini Lacy,CSHL Press)所述制备出转基因小鼠),就可采用标准技术评估ARCAP基因的表达。用Southern印迹分析或PCR技术进行最初的筛选以确定转基因是否已发生了整合。参见,例如,Sambrook等1989年所著的"Molecular Cloning,A Laboratory Manual"第二版,Cold Spring Harbor Press Plainview;NY,9.37-9.52章节中对Southern分析的描述。编码ARCAP蛋白的核酸在非人转基因动物组织中的表达可利用多种技术来评估,所述技术包括但不限于,对获自所述动物的组织样品进行Northern印迹分析,原位杂交分析和逆转录酶的PCR(RT-PCR)。
本发明的非人转基因动物可用作肝癌模型。这些动物尤其可用于鉴定对肝癌的治疗或预防有效的化合物或组合物。所述化合物或组合物可如下鉴定,即对本发明的非人转基因动物给药待测化合物或组合物,或使待测化合物或组合物与来源于所述非人转基因动物的器官,组织(例如肝脏)或细胞(例如肝细胞)接触。然后评价待测化合物或组合物对非人转基因动物,器官,组织或细胞的肝癌的影响。例如,可评估非人转基因动物中通过临床病理学鉴定的肿瘤的大小。能缓解癌症症状的待测化合物或组合物可有效治疗或预防肝癌。
可将待测化合物配制成药物组合物,方法是将所述化合物与药学上可接受的无毒性赋形剂或载体混合,然后通过任何给药途径将其给药于本发明的非人转基因动物。例如,可采用肠胃外给药途径,如皮下给药,肌肉内给药,血管内给药,皮内给药,鼻内给药,吸入给药,鞘内给药或腹膜内给药,以及肠道途径给药如舌下,口腔或直肠给药。
即使不作进一步的详述,相信本领域技术人员也能够在本文描述的基础上最完整地利用本发明。本文中所引用的出版物在此都以其全部内容纳入本文作参考。
其它实施方案
该说明书中公开的所有特征都可以以任何形式组合。本说明书所公开的每一特征都可以由达到相同,等价或类似目的的另一特征代替。因此,除非另有说明,本文所公开的每一特征仅仅是这类等价或类似特征中的一个实例。
从以上说明中,本领域技术人员能非常容易地发现本发明的特征,且能在不脱离本发明精神和范围的情况下,对本发明进行各种变动和修改,以使其适应各种用途和条件。因此,其它的实施方案也包括在以下权利要求的范围内。
序列表
<110>行政院退除役官兵辅导委员会台北荣民总医院
创盛基因科技股份有限公司
<120>表达雄激素受体复合物相关蛋白的转基因动物
<130>14321-003001
<140>US 10/206,566
<141>2002-07-2 5
<150>US 09/781,693
<151>2001-02-12
<150>US 60/262,312
<151>2001-01-17
<160>3
<170>FastSEQ for Windows Version 4.0
<210>1
<211>3340
<212>DNA
<213>小鼠(Mus musculus)
<220>
<221>CDS
<222>(218)...(2905)
<400>1
gagagtatga ggcgagccgt ggcccgggtg cctctccgct cccggggaga aggcgcggcc 60
gtggctgccg ccctctgagt cgcggcgccg gcgaggcccc ggggcgcgcg catggtgctg 120
gtgccgctcg ggtgttgatc ggcctgtccc ctccctctct tcccctcccc accccccgcg 180
gtggtctccc ctttcccacc ccagcccttg cggagcc atg gct cgg agt ggc tcc 235
Met Ala Arg Ser Gly Ser
1 5
tgc ccg cac ctg ttg tgg gac gtg agg aaa agg tcc ctt ggg ctg gag 283
Cys Pro His Leu Leu Trp Asp Val Arg Lys Arg Ser Leu Gly Leu Glu
10 15 20
gac ccg tcc cgg ctg agg agc cgc tac ctg gga aga aga gaa ttt atc 331
Asp Pro Ser Arg Leu Arg Ser Arg Tyr Leu Gly Arg Arg Glu Phe Ile
25 30 35
caa aga tta aaa ctt gaa gca act tta aat gtg cat gat ggc tgt gtt 379
Gln Arg Leu Lys Leu Glu Ala Thr Leu Asn Val His Asp Gly Cys Val
40 45 50
aat aca atc tgt tgg aat gac act gga gaa tat att tta tct ggc tct 427
Asn Thr Ile Cys Trp Asn Asp Thr Gly Glu Tyr Ile Leu Ser Gly Ser
55 60 65 70
gat gac act aaa ctt gta att agt aat cca tac agc aga aag gtt ttg 475
Asp Asp Thr Lys Leu Val Ile Ser Asn Pro Tyr Ser Arg Lys Val Leu
75 80 85
aca acc atc cgt tca ggg cat cga gca aat ata ttt agt gca aag ttt 523
Thr Thr Ile Arg Ser Gly His Arg Ala Asn Ile Phe Ser Ala Lys Phe
90 95 100
ttg ccg tgc aca gat gat aag cag att gtg tct tgc tct gga gat gga 571
Leu Pro Cys Thr Asp Asp Lys Gln Ile Val Ser Cys Ser Gly Asp Gly
105 110 115
gtc ata ttt tat act aac att gag caa gat gca gaa act aac aga cag 619
Val Ile Phe Tyr Thr Asn Ile Glu Gln Asp Ala Glu Thr Asn Arg Gln
120 125 130
tgc caa ttt acg tgc cat tat gga act act tat gag att atg act gta 667
Cys Gln Phe Thr Cys His Tyr Gly Thr Thr Tyr Glu Ile Met Thr Val
135 140 145 150
cca aac gac cct tac acc ttt ctg tcc tgt ggt gaa gat gga act gtt 715
Pro Asn Asp Pro Tyr Thr Phe Leu Ser Cys Gly Glu Asp Gly Thr Val
155 160 165
agg tgg ttt gac aca cgc atc aaa acc agt tgc aca aaa gaa gac tgt 763
Arg Trp Phe Asp Thr Arg Ile Lys Thr Ser Cys Thr Lys Glu Asp Cys
170 175 180
aaa gat gat att tta atc aac tgt agg cgt gct gcc aca tct gtg gct 811
Lys Asp Asp Ile Leu Ile Asn Cys Arg Arg Ala Ala Thr Ser Val Ala
185 190 195
att tgt ccc cca gta cca tat tac ctt gct gtg ggt tgt tct gac agc 859
Ile Cys Pro Pro Val Pro Tyr Tyr Leu Ala Val Gly Cys Ser Asp Ser
200 205 210
tca gta cgg att tat gat cgg cga atg ctg ggc aca aga gct aca ggg 907
Ser Val Arg Ile Tyr Asp Arg Arg Met Leu Gly Thr Arg Ala Thr Gly
215 220 225 230
aat tat gca ggc cga gga act act gga atg gtt gct cga ttt ata cct 955
Asn Tyr Ala Gly Arg Gly Thr Thr Gly Met Val Ala Arg Phe Ile Pro
235 240 245
tct cat ctt agt aac aaa tca tgc aga gtg aca tca ctg tgt tac agt 1003
Ser His Leu Ser Asn Lys Ser Cys Arg Val Thr Ser Leu Cys Tyr Ser
250 255 260
gaa gat ggt caa gag att ctt gtc agt tat tct tca gac tac atc tat 1051
Glu Asp Gly Gln Glu Ile Leu Val Ser Tyr Ser Ser Asp Tyr Ile Tyr
265 270 275
ctt ttt gac ccc aaa gat gat act gca cga gaa ctt aaa act cct tct 1099
Leu Phe Asp Pro Lys Asp Asp Thr Ala Arg Glu Leu Lys Thr Pro Ser
280 285 290
gca gag gag agg cga gaa gag tta cga cag cct cca gtt aag cgc ttg 1147
Ala Glu Glu Arg Arg Glu Glu Leu Arg Gln Pro Pro Val Lys Arg Leu
295 300 305 310
aga ctt cgt ggt gat tgg tca gat act ggt ccc aga gca cgg cca gaa 1195
Arg Leu Arg Gly Asp Trp Ser Asp Thr Gly Pro Arg Ala Arg Pro Glu
315 320 325
agt gaa cga gaa cga gat gga gag caa agt ccc aat gtg tca ctg atg 1243
Ser Glu Arg Glu Arg Asp Gly Glu Gln Ser Pro Asn Val Ser Leu Met
330 335 340
cag aga atg tct gat atg tta tca agg tgg ttt gaa gaa gca agt gaa 1291
Gln Arg Met Ser Asp Met Leu Ser Arg Trp Phe Glu Glu Ala Ser Glu
345 350 355
gtt gca caa agc aac aga gga aga gga aga cct cgg ccc aga ggt gga 1339
Val Ala Gln Ser Asn Arg Gly Arg Gly Arg Pro Arg Pro Arg Gly Gly
360 365 370
aca aat cag cca gat gtt tca act ctt cct acg gtt cca tca agt cct 1387
Thr Asn Gln Pro Asp Val Ser Thr Leu Pro Thr Val Pro Ser Ser Pro
375 380 385 390
aat ttg gaa gtg tgt gaa act gca atg gat gta gac atg cca gct gca 1435
Asn Leu Glu Val Cys Glu Thr Ala Met Asp Val Asp Met Pro Ala Ala
395 400 405
ctt ctt cag cct tct aca tcc tct aca gat cca gtt cag gct cag gca 1483
Leu Leu Gln Pro Ser Thr Ser Ser Thr Asp Pro Val Gln Ala Gln Ala
410 415 420
gcc aca gcc gcc ata gaa agc cct cgt tcc agc tcg ttg ctg tct tgc 1531
Ala Thr Ala Ala Ile Glu Ser Pro Arg Ser Ser Ser Leu Leu Ser Cys
425 430 435
cca gac agt gaa ccg agg cag tct gtt gag gcg tct gga cac cat gca 1579
Pro Asp Ser Glu Pro Arg Gln Ser Val Glu Ala Ser Gly His His Ala
440 445 450
cat cat cag tca gat tct cct tct tct gtg gtt aac aaa cag ctg gga 1627
His His Gln Ser Asp Ser Pro Ser Ser Val Val Asn Lys Gln Leu Gly
455 460 465 470
tcc atg tca ctt gat gag caa cag gat aac agt aat gag agg ctg agc 1675
Ser Met Ser Leu Asp Glu Gln Gln Asp Asn Ser Asn Glu Arg Leu Ser
475 480 485
ccc aaa cca ggg aca ggt gaa ccc gtt tta agt ttg cac tac agc aca 1723
Pro Lys Pro Gly Thr Gly Glu Pro Val Leu Ser Leu His Tyr Ser Thr
490 495 500
gaa gga aca act aca agc aca ata aaa ctg aac ttt aca gat gaa tgg 1771
Glu Gly Thr Thr Thr Ser Thr Ile Lys Leu Asn Phe Thr Asp Glu Trp
505 510 515
agc agt aca gcc tca agt tcc aga gga aat ggg agc cat tgc aaa tct 1819
Ser Ser Thr Ala Ser Ser Ser Arg Gly Asn Gly Ser His Cys Lys Ser
520 525 530
gag ggt cag gaa gaa tgc ttg gtc cct ccg agc tct gtg cag cca ccg 1867
Glu Gly Gln Glu Glu Cys Leu Val Pro Pro Ser Ser Val Gln Pro Pro
535 540 545 550
gaa gga gac agt gaa aca aga gct cct gaa gaa cta tca gag aaa gga 1915
Glu Gly Asp Ser Glu Thr Arg Ala Pro Glu Glu Leu Ser Glu Lys Gly
555 560 565
aca ctt cca gaa aac ctc act caa aac cag ata gat aca gca caa ctt 1963
Thr Leu Pro Glu Asn Leu Thr Gln Asn Gln Ile Asp Thr Ala Gln Leu
570 575 580
gat aac ttc cca gct gag cca ttg gat tct aac tca gga gag aag aat 2011
Asp Asn Phe Pro Ala Glu Pro Leu Asp Ser Asn Ser Gly Glu Lys Asn
585 590 595
aac cca agt cag gac agc cct tgt ggg ctt cca gaa gaa ggc act ttg 2059
Asn Pro Ser Gln Asp Ser Pro Cys Gly Leu Pro Glu Glu Gly Thr Leu
600 605 610
tct gaa aca gac agg gag act tgt gag cag gcc agc act gag agt gct 2107
Ser Glu Thr Asp Arg Glu Thr Cys Glu Gln Ala Ser Thr Glu Ser Ala
615 620 625 630
acc agg cat gct agc acc aag cct gaa ctc cca tcc cag aca gaa gcc 2155
Thr Arg His Ala Ser Thr Lys Pro Glu Leu Pro Ser Gln Thr Glu Ala
635 640 645
att gag cag gcc agc act gag agt gct acc agg cat acc agt gcc aat 2203
Ile Glu Gln Ala Ser Thr Glu Ser Ala Thr Arg His Thr Ser Ala Asn
650 655 660
cct gaa ctc cca tcc cag aca gaa gcc ata gca cct tta gct cat gaa 2251
Pro Glu Leu Pro Ser Gln Thr Glu Ala Ile Ala Pro Leu Ala His Glu
665 670 675
gac cca tct gcc agg gac tct gct ctc cag gac aca gat gac agc gat 2299
Asp Pro Ser Ala Arg Asp Ser Ala Leu Gln Asp Thr Asp Asp Ser Asp
680 685 690
gat gat ccg gtc ttg atc cct ggt gca aga tac cga aca gga cct ggt 2347
Asp Asp Pro Val Leu Ile Pro Gly Ala Arg Tyr Arg Thr Gly Pro Gly
695 700 705 710
gat aga cgc tcc gct gtt gcc cgc att cag gag ttc ttc agg agg aga 2395
Asp Arg Arg Ser Ala Val Ala Arg Ile Gln Glu Phe Phe Arg Arg Arg
715 720 725
aaa gaa agg aaa gaa atg gaa gag ctg gat act ttg aac att agg agg 2443
Lys Glu Arg Lys Glu Met Glu Glu Leu Asp Thr Leu Asn Ile Arg Arg
730 735 740
cca cta gta aag atg gtt tat aag ggc cac cgc aac tcc cgg aca atg 2491
Pro Leu Val Lys Met Val Tyr Lys Gly His Arg Asn Ser Arg Thr Met
745 750 755
ata aaa gaa gcc aat ttc tgg ggt gct aac ttt gta atg agc ggt tcc 2539
Ile Lys Glu Ala Asn Phe Trp Gly Ala Asn Phe Val Met Ser Gly Ser
760 765 770
gat tgt ggc cat atc ttc atc tgg gac cgg cac act gcg gag cat ttg 2587
Asp Cys Gly His Ile Phe Ile Trp Asp Arg His Thr Ala Glu His Leu
775 780 785 790
atg ctt ctg gaa gct gat aat cat gtg gtc aac tgc ctg cag ccc cat 2635
Met Leu Leu Glu Ala Asp Asn His Val Val Asn Cys Leu Gln Pro His
795 800 805
ccg ttt gac cca att cta gcc tca tct ggc ata gat tat gac ata aag 2683
Pro Phe Asp Pro Ile Leu Ala Ser Ser Gly Ile Asp Tyr Asp Ile Lys
810 815 820
atc tgg tcg cca cta gaa gag tca aga att ttt aat cga aaa ctt gct 2731
Ile Trp Ser Pro Leu Glu Glu Ser Arg Ile Phe Asn Arg Lys Leu Ala
825 830 835
gat gaa gtt ata act cgg aat gaa ctc acg ctg gaa gag act cgg aac 2779
Asp Glu Val Ile Thr Arg Asn Glu Leu Thr Leu Glu Glu Thr Arg Asn
840 845 850
acc atc acc gtc cca gcc tct ttc atg ttg agg atg ttg gcg tca ctg 2827
Thr Ile Thr Val Pro Ala Ser Phe Met Leu Arg Met Leu Ala Ser Leu
855 860 865 870
aat cat atc cga gct gac cgt ctg gag ggt gac aga tca gaa ggt tca 2875
Asn His Ile Arg Ala Asp Arg Leu Glu Gly Asp Arg Ser Glu Gly Ser
875 880 885
ggt cag gag aat gaa aat gag gat gaa gaa taaagaactc cttggcaagc 2925
Gly Gln Glu Asn Glu Asn Glu Asp Glu Glu
890 895
acttagatgt tctgagattt gtatacgaca tttattatat tttttttctt tacagaactt 2985
tagtgcaatt taaggctatg ggtttttttt tttctttttt tttggagttc ttccctattt 3045
tggggataac caaacattgg tttggaatga gtgtgtgcat gagttgggag agtgtgtaaa 3105
acaaagtaag caaaatgttt tttgaaacct tttgccgtgt atggagtccc aaaaaaaaaa 3165
aaaaaaaaaa aaaaaagcaa agtgcaatac ttcctgaccc tccgctgtgg gagcttggat 3225
caatgctgaa gtcattttca ttgtagtgaa aacgttggtt caaataaatt tctacacttg 3285
ccatttgcaa aaaaaaaaaa aaaaaaaaaa aaaaaaagcg gccgctgaat tctag 3340
<210>2
<211>2688
<212>DNA
<213>小鼠(Mus musculus)
<400>2
atggctcgga gtggctcctg cccgcacctg ttgtgggacg tgaggaaaag gtcccttggg 60
ctggaggacc cgtcccggct gaggagccgc tacctgggaa gaagagaatt tatccaaaga 120
ttaaaacttg aagcaacttt aaatgtgcat gatggctgtg ttaatacaat ctgttggaat 180
gacactggag aatatatttt atctggctct gatgacacta aacttgtaat tagtaatcca 240
tacagcagaa aggttttgac aaccatccgt tcagggcatc gagcaaatat atttagtgca 300
aagtttttgc cgtgcacaga tgataagcag attgtgtctt gctctggaga tggagtcata 360
ttttatacta acattgagca agatgcagaa actaacagac agtgccaatt tacgtgccat 420
tatggaacta cttatgagat tatgactgta ccaaacgacc cttacacctt tctgtcctgt 480
ggtgaagatg gaactgttag gtggtttgac acacgcatca aaaccagttg cacaaaagaa 540
gactgtaaag atgatatttt aatcaactgt aggcgtgctg ccacatctgt ggctatttgt 600
cccccagtac catattacct tgctgtgggt tgttctgaca gctcagtacg gatttatgat 660
cggcgaatgc tgggcacaag agctacaggg aattatgcag gccgaggaac tactggaatg 720
gttgctcgat ttataccttc tcatcttagt aacaaatcat gcagagtgac atcactgtgt 780
tacagtgaag atggtcaaga gattcttgtc agttattctt cagactacat ctatcttttt 840
gaccccaaag atgatactgc acgagaactt aaaactcctt ctgcagagga gaggcgagaa 900
gagttacgac agcctccagt taagcgcttg agacttcgtg gtgattggtc agatactggt 960
cccagagcac ggccagaaag tgaacgagaa cgagatggag agcaaagtcc caatgtgtca 1020
ctgatgcaga gaatgtctga tatgttatca aggtggtttg aagaagcaag tgaagttgca 1080
caaagcaaca gaggaagagg aagacctcgg cccagaggtg gaacaaatca gccagatgtt 1140
tcaactcttc ctacggttcc atcaagtcct aatttggaag tgtgtgaaac tgcaatggat 1200
gtagacatgc cagctgcact tcttcagcct tctacatcct ctacagatcc agttcaggct 1260
caggcagcca cagccgccat agaaagccct cgttccagct cgttgctgtc ttgcccagac 1320
agtgaaccga ggcagtctgt tgaggcgtct ggacaccatg cacatcatca gtcagattct 1380
ccttcttctg tggttaacaa acagctggga tccatgtcac ttgatgagca acaggataac 1440
agtaatgaga ggctgagccc caaaccaggg acaggtgaac ccgttttaag tttgcactac 1500
agcacagaag gaacaactac aagcacaata aaactgaact ttacagatga atggagcagt 1560
acagcctcaa gttccagagg aaatgggagc cattgcaaat ctgagggtca ggaagaatgc 1620
ttggtccctc cgagctctgt gcagccaccg gaaggagaca gtgaaacaag agctcctgaa 1680
gaactatcag agaaaggaac acttccagaa aacctcactc aaaaccagat agatacagca 1740
caacttgata acttcccagc tgagccattg gattctaact caggagagaa gaataaccca 1800
agtcaggaca gcccttgtgg gcttccagaa gaaggcactt tgtctgaaac agacagggag 1860
acttgtgagc aggccagcac tgagagtgct accaggcatg ctagcaccaa gcctgaactc 1920
ceatcccaga cagaagccat tgagcaggcc agcactgaga gtgctaccag gcataccagt 1980
gccaatcctg aactcccatc ccagacagaa gccatagcac ctttagctca tgaagaccea 2040
tctgccaggg actctgctct ccaggacaca gatgacagcg atgatgatcc ggtcttgatc 2100
cctggtgcaa gataccgaac aggacctggt gatagacgct ccgctgttgc ccgcattcag 2160
gagttcttca ggaggagaaa agaaaggaaa gaaatggaag agctggatac tttgaacatt 2220
aggaggccac tagtaaagat ggtttataag ggccaccgca actcccggac aatgataaaa 2280
gaagccaatt tctggggtgc taactttgta atgagcggtt ccgattgtgg ccatatcttc 2340
atctgggacc ggcacactgc ggagcatttg atgcttctgg aagctgataa tcatgtggtc 2400
aactgcctgc agccccatcc gtttgaccca attctagcct catctggcat agattatgac 2460
ataaagatct ggtcgccact agaagagtca agaattttta atcgaaaact tgctgatgaa 2520
gttataactc ggaatgaact cacgctggaa gagactcgga acaccatcac cgtcccagcc 2580
tctttcatgt tgaggatgtt ggcgtcactg aatcatatcc gagctgaccg tctggagggt 2640
gacagatcag aaggttcagg tcaggagaat gaaaatgagg atgaagaa 2688
<210>3
<211>896
<212>PRT
<213>小鼠(Mus musculus)
<400>3
Met Ala Arg Ser Gly Ser Cys Pro His Leu Leu Trp Asp Val Arg Lys
1 5 10 15
Arg Ser Leu Gly Leu Glu Asp Pro Ser Arg Leu Arg Ser Arg Tyr Leu
20 25 30
Gly Arg Arg Glu Phe Ile Gln Arg Leu Lys Leu Glu Ala Thr Leu Asn
35 40 45
Val His Asp Gly Cys Val Asn Thr Ile Cys Trp Asn Asp Thr Gly Glu
50 55 60
Tyr Ile Leu Ser Gly Ser Asp Asp Thr Lys Leu Val Ile Ser Asn Pro
65 70 75 80
Tyr Ser Arg Lys Val Leu Thr Thr Ile Arg Ser Gly His Arg Ala Asn
85 90 95
Ile Phe Ser Ala Lys Phe Leu Pro Cys Thr Asp Asp Lys Gln Ile Val
100 105 110
Ser Cys Ser Gly Asp Gly Val Ile Phe Tyr Thr Asn Ile Glu Gln Asp
115 120 125
Ala Glu Thr Asn Arg Gln Cys Gln Phe Thr Cys His Tyr Gly Thr Thr
130 135 140
Tyr Glu Ile Met Thr Val Pro Asn Asp Pro Tyr Thr Phe Leu Ser Cys
145 150 155 160
Gly Glu Asp Gly Thr Val Arg Trp Phe Asp Thr Arg Ile Lys Thr Ser
165 170 175
Cys Thr Lys Glu Asp Cys Lys Asp Asp Ile Leu Ile Asn Cys Arg Arg
180 185 190
Ala Ala Thr Ser Val Ala Ile Cys Pro Pro Val Pro Tyr Tyr Leu Ala
195 200 205
Val Gly Cys Ser Asp Ser Ser Val Arg Ile Tyr Asp Arg Arg Met Leu
210 215 220
Gly Thr Arg Ala Thr Gly Asn Tyr Ala Gly Arg Gly Thr Thr Gly Met
225 230 235 240
Val Ala Arg Phe Ile Pro Ser His Leu Ser Asn Lys Ser Cys Arg Val
245 250 255
Thr Ser Leu Cys Tyr Ser Glu Asp Gly Gln Glu Ile Leu Val Ser Tyr
260 265 270
Ser Ser Asp Tyr Ile Tyr Leu Phe Asp Pro Lys Asp Asp Thr Ala Arg
275 280 285
Glu Leu Lys Thr Pro Ser Ala Glu Glu Arg Arg Glu Glu Leu Arg Gln
290 295 300
Pro Pro Val Lys Arg Leu Arg Leu Arg Gly Asp Trp Ser Asp Thr Gly
305 310 315 320
Pro Arg Ala Arg Pro Glu Ser Glu Arg Glu Arg Asp Gly Glu Gln Ser
325 330 335
Pro Asn Val Ser Leu Met Gln Arg Met Ser Asp Met Leu Ser Arg Trp
340 345 350
Phe Glu Glu Ala Ser Glu Val Ala Gln Ser Asn Arg Gly Arg Gly Arg
355 360 365
Pro Arg Pro Arg Gly Gly Thr Asn Gln Pro Asp Val Ser Thr Leu Pro
370 375 380
Thr Val Pro Ser Ser Pro Asn Leu Glu Val Cys Glu Thr Ala Met Asp
385 390 395 400
Val Asp Met Pro Ala Ala Leu Leu Gln Pro Ser Thr Ser Ser Thr Asp
405 410 415
Pro Val Gln Ala Gln Ala Ala Thr Ala Ala Ile Glu Ser Pro Arg Ser
420 425 430
Ser Ser Leu Leu Ser Cys Pro Asp Ser Glu Pro Arg Gln Ser Val Glu
435 440 445
Ala Ser Gly His His Ala His His Gln Ser Asp Ser Pro Ser Ser Val
450 455 460
Val Asn Lys Gln Leu Gly Ser Met Ser Leu Asp Glu Gln Gln Asp Asn
465 470 475 480
Ser Asn Glu Arg Leu Ser Pro Lys Pro Gly Thr Gly Glu Pro Val Leu
485 490 495
Ser Leu His Tyr Ser Thr Glu Gly Thr Thr Thr Ser Thr Ile Lys Leu
500 505 510
Asn Phe Thr Asp Glu Trp Ser Ser Thr Ala Ser Ser Ser Arg Gly Asn
515 520 525
Gly Ser His Cys Lys Ser Glu Gly Gln Glu Glu Cys Leu Val Pro Pro
530 535 540
Ser Ser Val Gln Pro Pro Glu Gly Asp Ser Glu Thr Arg Ala Pro Glu
545 550 555 560
Glu Leu Ser Glu Lys Gly Thr Leu Pro Glu Asn Leu Thr Gln Asn Gln
565 570 575
Ile Asp Thr Ala Gln Leu Asp Asn Phe Pro Ala Glu Pro Leu Asp Ser
580 585 590
Asn Ser Gly Glu Lys Asn Asn Pro Ser Gln Asp Ser Pro Cys Gly Leu
595 600 605
Pro Glu Glu Gly Thr Leu Ser Glu Thr Asp Arg Glu Thr Cys Glu Gln
610 615 620
Ala Ser Thr Glu Ser Ala Thr Arg His Ala Ser Thr Lys Pro Glu Leu
625 630 635 640
Pro Ser Gln Thr Glu Ala Ile Glu Gln Ala Ser Thr Glu Ser Ala Thr
645 650 655
Arg His Thr Ser Ala Asn Pro Glu Leu Pro Ser GlnThr Glu Ala Ile
660 665 670
Ala Pro Leu Ala His Glu Asp Pro Ser Ala Arg Asp Ser Ala Leu Gln
675 680 685
Asp Thr Asp Asp Ser Asp Asp Asp Pro Val Leu Ile Pro Gly Ala Arg
690 695 700
Tyr Arg Thr Gly Pro Gly Asp Arg Arg Ser Ala Val Ala Arg Ile Gln
705 710 715 720
Glu Phe Phe Arg Arg Arg Lys Glu Arg Lys Glu Met Glu Glu Leu Asp
725 730 735
Thr Leu Asn Ile Arg Arg Pro Leu Val Lys Met Val Tyr Lys Gly His
740 745 750
Arg Asn Ser Arg Thr Met Ile Lys Glu Ala Asn Phe Trp Gly Ala Asn
755 760 765
Phe Val Met Ser Gly Ser Asp Cys Gly His Ile Phe Ile Trp Asp Arg
770 775 780
His Thr Ala Glu His Leu Met Leu Leu Glu Ala Asp Asn His Val Val
785 790 795 800
Asn Cys Leu Gln Pro His Pro Phe Asp Pro Ile Leu Ala Ser Ser Gly
805 810 815
Ile Asp Tyr Asp Ile Lys Ile Trp Ser Pro Leu Glu Glu Ser Arg Ile
820 825 830
Phe Asn Arg Lys Leu Ala Asp Glu Val Ile Thr Arg Asn Glu Leu Thr
835 840 845
Leu Glu Glu Thr Arg Asn Thr Ile Thr Val Pro Ala Ser Phe Met Leu
850 855 860
Arg Met Leu Ala Ser Leu Asn His Ile Arg Ala Asp Arg Leu Glu Gly
865 870 875 880
Asp Arg Ser Glu Gly Ser Gly Gln Glu Asn Glu Asn Glu Asp Glu Glu
885 890 895
Claims (50)
1.一种基本纯的多肽,其包含与SEQ ID NO:3至少70%相同的氨基酸序列,其中所述多肽与雄激素受体结合并增强雄激素受体反式激活雄激素应答基因的能力。
2.权利要求1的多肽,其中所述氨基酸序列与SEQ ID NO:3至少80%相同。
3.权利要求2的多肽,其中所述氨基酸序列与SEQ ID NO:3至少90%相同。
4.权利要求3的多肽,其中所述氨基酸序列与SEQ ID NO:3至少95%相同。
5.权利要求4的多肽,其中所述氨基酸序列是SEQ ID NO:3。
6.一种基本纯的多肽,其包含具有多至30个保守氨基酸取代的SEQ ID NO:3的氨基酸序列,该多肽能与雄激素受体结合,并增强雄激素受体反式激活雄激素应答基因的能力。
7.一种基本纯的由在严谨条件下与具有SEQ ID NO:2所示序列的探针杂交的核酸编码的多肽,其中所述多肽能与雄激素受体结合,并增强雄激素受体反式激活雄激素应答基因的能力。
8.一种分离的核酸,其编码权利要求1的多肽。
9.权利要求8的核酸,其中所述氨基酸序列是SEQ ID NO:3。
10.编码权利要求6的多肽的分离的核酸。
11.一种分离的核酸,其包含在严谨条件下能与单链探针杂交的链,该探针的序列为SEQ ID NO:2所示序列或SEQ ID NO:2序列的互补体。
12.权利要求11的核酸,其中所述核酸编码多肽,该多肽能与雄激素受体结合,并增强雄激素受体反式激活雄激素应答基因的能力。
13.权利要求12的核酸,其中所述多肽的氨基酸序列包括SEQ IDNO:3。
14.权利要求11的核酸,其中所述链的长度至少是15个核苷酸。
15.包含权利要求8的核酸的载体。
16.权利要求15的载体,其中所述氨基酸序列是SEQ ID NO:3。
17.包含权利要求10的核酸的载体。
18.包含权利要求11的核酸的载体。
19.权利要求18的载体,其中所述核酸编码多肽,该多肽能与雄激素受体结合,并增强雄激素受体反式激活雄激素应答基因的能力。
20.包含权利要求8的核酸的细胞。
21.权利要求20的细胞,其中所述氨基酸序列是SEQ ID NO:3。
22.包含权利要求10的核酸的细胞。
23.包含权利要求11的核酸的细胞。
24.权利要求23的细胞,其中所述核酸编码多肽,该多肽能与雄激素受体结合,并增强雄激素受体反式激活雄激素应答基因的能力。
25.一种在其基因组中包含转基因的转基因动物,所述转基因包含权利要求8的核酸,该转基因动物与野生型动物相比对雄激素的应答增强。
26.权利要求25的转基因动物,其中所述动物是啮齿类动物。
27.权利要求26的转基因动物,其中所述动物是小鼠。
28.权利要求25的转基因动物,其中所述氨基酸序列是SEQ IDNO:3。
29.权利要求28的转基因动物,其中所述动物是啮齿类动物。
30.权利要求29的转基因动物,其中所述动物是小鼠。
31.一种在其基因组中包含转基因的转基因动物,所述转基因包含权利要求10的核酸,该转基因动物与野生型动物相比对雄激素的应答增强。
32.权利要求31的转基因动物,其中所述动物是啮齿类动物。
33.权利要求32的转基因动物,其中所述动物是小鼠。
34.一种在其基因组中包含转基因的转基因动物,所述转基因包含权利要求11的核酸,该转基因动物与野生型动物相比对雄激素的应答增强。
35.权利要求34的转基因动物,其中所述动物是啮齿类动物。
36.权利要求35的转基因动物,其中所述动物是小鼠。
37.一种制备多肽的方法,其包括培养权利要求20的细胞,在该细胞中表达所述多肽,和从培养物中分离这种多肽。
38.一种制备转录物的方法,包括将权利要求11的核酸导入到细胞中,从该核酸制备转录物。
39.一种分离的抗体,其能与包含SEQ ID NO:3所示氨基酸序列的多肽结合。
40.一种确定动物样品中是否包含癌细胞的方法,该方法包括:
提供来自动物的样品,和
检测样品中雄激素受体复合物相关蛋白的基因的表达水平,样品中雄激素受体复合物相关蛋白基因的表达水平高于正常样品中的水平时,指示该动物样品中包含癌细胞。
41.权利要求40的方法,其中所述样品是血样品。
42.权利要求40的方法,其中所述癌细胞是肝癌细胞。
43.权利要求40的方法,其中所述动物是啮齿类动物。
44.权利要求43的方法,其中所述动物是小鼠。
45.一种治疗动物癌症的方法,其包括:
鉴定含有癌细胞的动物,所述癌细胞表达雄激素受体复合物相关蛋白基因,以及
用能阻断雄激素受体复合物相关蛋白与雄激素受体的结合的组合物或能减弱雄激素受体反式激活雄激素应答基因的能力的组合物治疗所述动物。
46.权利要求45的方法,其中所述组合物包含权利要求39的抗体。
47.权利要求45的方法,其中所述组合物包含权利要求11的核酸。
48.权利要求45的方法,其中所述癌症是肝癌。
49.权利要求45的方法,其中所述动物是啮齿类动物。
50.权利要求49的方法,其中所述动物是小鼠。
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US10/206,566 | 2002-07-25 | ||
US10/206,566 US20030082721A1 (en) | 2001-01-17 | 2002-07-25 | Transgenic animals expressing androgen receptor complex-associated protein |
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US (1) | US20030082721A1 (zh) |
EP (1) | EP1398323A1 (zh) |
KR (1) | KR20040010402A (zh) |
CN (1) | CN1495195A (zh) |
BR (1) | BR0302594A (zh) |
RU (1) | RU2333959C2 (zh) |
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Cited By (1)
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CN109247302A (zh) * | 2017-07-14 | 2019-01-22 | 张泰阶 | 人雄激素受体复合物相关蛋白转基因小鼠 |
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EP2818478B1 (en) * | 2011-10-28 | 2017-02-01 | Regeneron Pharmaceuticals, Inc. | Humanized IL-6 and IL-6 receptor |
WO2020140858A1 (en) * | 2019-01-03 | 2020-07-09 | Chen Show Li | Method for treating amyotrophic lateral sclerosis |
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US5789170A (en) * | 1996-05-23 | 1998-08-04 | Wisconsin Alumni Research Foundation | Specific co-activator for human androgen receptor |
US6525187B1 (en) * | 1998-07-31 | 2003-02-25 | Diagnostic Products Corporation | Polynucleotide encoding autoantigens associated with endometriosis |
EP1196577A2 (en) * | 1999-07-21 | 2002-04-17 | Incyte Genomics, Inc. | Cell cycle and proliferation proteins |
US20030099976A1 (en) * | 2001-01-17 | 2003-05-29 | Tai-Jay Chang | Androgen receptor complex-associated protein |
US6974683B2 (en) * | 2001-01-17 | 2005-12-13 | Veterans General Hospital | Nucleic acid encoding androgen receptor complex-associated protein |
US6783969B1 (en) * | 2001-03-05 | 2004-08-31 | Nuvelo, Inc. | Cathepsin V-like polypeptides |
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2002
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US20030082721A1 (en) | 2003-05-01 |
BR0302594A (pt) | 2004-08-24 |
KR20040010402A (ko) | 2004-01-31 |
TWI344988B (en) | 2011-07-11 |
RU2333959C2 (ru) | 2008-09-20 |
RU2003123598A (ru) | 2005-02-10 |
TW200407433A (en) | 2004-05-16 |
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