CN108192969B - 一种辅助癫痫诊断的标志物及其检测试剂盒 - Google Patents
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Abstract
本发明公开了一种辅助癫痫诊断的标志物及其检测试剂盒,涉及癫痫诊断领域。本发明的研究通过对一个癫痫家系患者的成员的FASN基因的检测发现,该家系中患有癫痫的成员的FASN基因上具有突变c.G2998A,该基因突变导致其对应的FASN蛋白的第1000位氨基酸由天冬氨基酸(D)变异为天冬酰胺(N),为错义突变。含有c.G2998A突变的核酸分子,或者含有突变p.D1000N的蛋白均可以作为癫痫诊断的生物标志物,从分子水平上为癫痫的诊断提供一种快速、可靠的检测辅助标记物。
Description
技术领域
本发明涉及癫痫诊断领域,具体而言,涉及一种辅助癫痫诊断的标志物及其检测试剂盒。
背景技术
癫痫是慢性反复发作性短暂脑功能失调综合征。以脑部神经元高度同步化异常放电引起反复痫性发作为特征。癫痫是神经***常见疾病之一,患病率仅次于脑卒中。癫痫的发病率与年龄有关。一般认为1岁以内患病率最高,1~10岁以后逐渐降低。我国癫痫患者男女之比为(1.15-1.7):1。
癫痫病因极其复杂,可分三大类,并存在多种影响发病的因素:
1.特发性癫痫
可疑遗传倾向且无其他明显病因,常在特定年龄段起病,有特征性临床及脑电图表现,诊断较明确。
2.症状性癫痫
中枢神经***病变影响结构或功能等,如染色体异常、局灶性或弥漫性脑部疾病,以及某些***性疾病所致。
(1)局限性或弥漫性脑部疾病①先天性异常胚胎发育中各种病因导致脑穿通畸形、小头畸形、先天性脑积水胼胝体缺如及大脑皮质发育不全,围生期胎儿脑损伤等;②获得性脑损伤如脑外伤,颅脑手术后,脑卒中后,颅内感染后,急性酒精中毒;③产伤所致新生儿癫痫发生率约为1%,分娩时合并产伤多伴脑出血或脑缺氧损害,新生儿合并脑先天发育畸形或产伤,癫痫发病率高达25%;④感染包括中枢神经***细菌、病毒、真菌、寄生虫、螺旋体感染及AIDS神经***并发症等;⑤脑血管疾病如脑动静脉畸形、脑梗死和脑出血等;⑥颅内原发性肿瘤如神经胶质瘤、脑膜瘤等;⑦遗传代谢性疾病如结节性硬化、脑-面血管瘤病、苯丙酮酸尿症等;⑧神经***变性疾病如Alzheimer病、Pick病等约1/3的患者合并癫痫发作。
(2)***性疾病①缺氧性脑病如心搏骤停、CO中毒窒息、麻醉意外和呼吸衰竭等可引起肌阵挛性发作或全身性大发作;②代谢性脑病如低血糖症,其他代谢及内分泌障碍如高血糖症、低钙血症、低钠血症,以及***、肝性脑病和甲状腺毒血症等均可导致癫痫发作;③心血管疾病如心脏骤停、高血压脑病等;④热性惊厥热性发作;⑤子痫;⑥中毒如酒精、异烟肼、卡巴唑等药物及铅、铊等重金属中毒。
3.隐源性癫痫
较多见,临床表现提示症状性癫痫,但未找到明确病因,可在特殊年龄段起病,无特定临床和脑电图表现。
癫痫的临床表现有以下:
1.全面强直-阵挛发作(大发作)
系指全身肌肉抽动及意识丧失的发作。以产伤、脑外伤、脑瘤等较常见。强直-阵挛发作可发生在任何年龄,是各种癫痫中最常见的发作类型。其典型发作可分为强直期、阵挛期、恢复期三个临床阶段。发作期间脑电图为典型的爆发性多棘波和棘-慢波综合,每次棘-慢波综合可伴有肌肉跳动。
2.单纯部分发作
是指脑的局部皮质异常放电而引起的与该部位的功能相对应的症状,发作时意识始终存在,包括运动、感觉、自主神经、精神症状及体征。分为四组:①伴运动症状者;②伴躯体感觉或特殊感觉症状者;③伴自主神经症状和体征者;④伴精神症状者。
3.复杂部分发作
习惯上又称精神运动性发作,伴有意识障碍。先兆多在意识丧失前或即将丧失时发生,发作后不能或部分不能回忆发作细节。4.失神发作(小发作)
其典型表现为意识障碍,而不伴先兆或发作后症状。
5.癫痫持续状态
是指单次癫痫发作超过5分钟,或者短期内癫痫频繁发作,以致患者尚未从前一次发作中完全恢复而又有另一次发作。癫痫持续状态是一种需要抢救的急症。
癫痫的治疗可分为控制发作、病因治疗、外科治疗、一般卫生及预防五个方面。其中最重要的是控制发作,目前以药物治疗为主。
临床上根据癫痫发作类型选用抗癫痫药物,一旦找到可以完全控制发作的药物和剂量,就应不间断地应用。一般应于发作完全控制后,如无不良反应再继续服用3~5年,方可考虑逐渐停药。目前多主张用单药治疗,确认单药治疗失败后,方可加用第2种药物。如失神发作或肌阵挛发作无法用单药控制者,可合用乙琥胺和丙戊酸钠,或其一加用苯二氮类。对合并多种发作类型者可以根据发作类型合理联合用药,但以不超过3种药物为宜。
用药宜从小剂量开始,然后逐渐增量,以既能控制发作,又不产生毒性反应的最小有效剂量为宜。换药宜采取先加用新药及后递减旧药的原则。不能骤然停药。
有些器质性脑病的癫痫患者可能需要终身服药;有人主张发病年龄大于30岁者需谨慎停药,因其停药后复发率较高,需长期服药或终身服药。但仍有10%~15%患者难以控制发作,可以采用外科治疗。
在预防方面:
1.预防癫痫病的发生,应详细地进行家系调查,了解患者双亲同胞和近亲中是否有癫痫发作及其发作特点,对能引起智力低下和癫痫的一些严重遗传性疾病,应进行产前诊断或新生儿期过筛检查,以决定终止妊娠或早期进行治疗。防止分娩意外,新生儿产伤是癫痫发病的重要原因之一,避免产伤对预防癫痫有重要意义。
2.对癫痫患者要及时诊断,及早治疗,治疗越早脑损伤越小,复发越少,预后越好。去除或减轻引起癫痫的原发病如颅内占位性疾病、代谢异常、感染等,对反复发作的病例也有重要意义。
3.癫痫是一种慢性疾病,可迁延数年、甚至数十年之久,因而可对患者身体、精神、婚姻以及社会经济地位等,造成严重的不良影响。患者在家庭关系、学校教育和就业等方面的不幸和挫折、文体活动方面的限制等,可使患者产生耻辱感和悲观心理,从而严重影响患者的身心发育,这就要求社会各界对癫痫患者给予理解和支持。
早诊断早治疗是防治癫痫的有效方法,但目前缺乏癫痫诊断相关的标志物。
鉴于此,特提出本发明。
发明内容
本发明的目的在于提供一种用于辅助癫痫诊断的生物标志物,该生物标志物可以用于辅助癫痫的诊断,为癫痫的诊断提供一种新的思路。
本发明的另一目的在于提供上述生物标志物的应用。
本发明的另一目的在于提供一种辅助癫痫诊断的试剂盒。
本发明的另一目的在于提供一种用于辅助癫痫诊断的基因芯片。
本发明是这样实现的:
第一方面,本发明提供了一种辅助癫痫诊断的生物标志物,其为核酸分子,其相对于正常FASN的编码序列具有突变c.G2998A。
FASN基因编码的酶是一种多功能蛋白,该蛋白的主要功能是催化乙酰辅酶A和丙二酰辅酶A合成棕榈酸酯,在NADPH存在下,最终生成长链饱和脂肪酸。在一些癌细胞系中,该蛋白的N端能与***受体-α(ER-α)的C端结合。
进一步地,在本发明的一些实施方案中,所述标志物的碱基序列如SEQ ID NO.1所示。
本发明的研究通过对一个癫痫家系患者的成员的FASN的检测测序发现,该家系中患有癫痫的成员的FASN上具有突变c.G2998A,该基因突变导致其对应的FASN蛋白的第1000位氨基酸由天冬氨基酸(D)变异为天冬酰胺(N),为错义突变。该变异不属于多态性位点,在人群中发生频率极低。在HGMD专业版数据库中未见报道。经家系验证分析,受检人(图1中3号成员)之父(图1中2号成员)该位点无变异,受检人之母(图1中1号成员)该位点杂合变异。
因此,含有c.G2998A突变的核酸分子即相对于正常FASN的编码序列具有突变c.G2998A的核酸片段,或者含有突变p.D1000N的蛋白均可以作为癫痫诊断的生物标志物,从分子水平上为癫痫的诊断提供一种快速、可靠的检测辅助标记物。
第二方面,本发明提供了一种辅助癫痫诊断的生物标志物,其为蛋白或多肽,其相对于正常FASN蛋白具有突变p.D1000N。
进一步地,在本发明的一些实施方案中,所述标志物的氨基酸序列如SEQ ID NO.2所示。
另一方面,本发明提供了检测第一方面所述的标志物的引物或探针、或检测权第二方面所述的生物标志物的抗体在制备癫痫辅助诊断试剂盒、癫痫产前诊断试剂盒或新生儿癫痫筛查试剂盒中的应用。
基于本发明的研究成果,本领域技术人员应当很容易将用于检测含有c.G2998A突变的FASN突变基因的引物或探针,以及用于检测含有p.D1000N突变的FASN突变蛋白的抗体用于制备癫痫辅助诊断试剂盒、癫痫产前诊断试剂盒以及新生儿癫痫筛查试剂盒等领域中。
或者将检测含有c.G2998A突变的FASN突变基因的探针用于制备辅助癫痫诊断的基因芯片中。
进一步,在本发明的一些实施方案中,所述探针相对于正常FASN的编码序列的第2998位的碱基为A。
另一方面,本发明提供了一种辅助癫痫诊断的试剂盒,其含有用于检测第一方面所述的生物标志物的引物或探针;
或者所述试剂盒含有检测第二方面所述的生物标志物的抗体。
另一方面,本发明提供了一种用于辅助癫痫诊断的基因芯片,该基因芯片上含有用于与目标片段特异性结合的探针,所述目标片段性相对于正常FASN的编码序列具有突变c.G2998A。
所述探针与目标片段特异性结合的区域涵盖如下位置:相对于正常FASN的第2998位。
进一步,在本发明的一些实施方案中,所述探针相对于正常FASN编码序列的第2998位的碱基为A。
进一步,在本发明的一些实施方案中,所述目标片段为来自待测主体的FASN的编码序列。
进一步,在本发明的一些实施方案中,所述主体为人。
附图说明
为了更清楚地说明本发明实施例的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,应当理解,以下附图仅示出了本发明的某些实施例,因此不应被看作是对范围的限定,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他相关的附图。
图1为本发明实施例1中的受检对象癫痫患者家系的遗传家系图谱;
图2-A为本发明实施例1中的遗传家系图谱中的3号成员的FASN测序结果图;
图2-B为本发明实施例1中的遗传家系图谱中的1号成员的FASN测序结果图;
图2-C为本发明实施例1中的遗传家系图谱中的2号成员的FASN测序结果图。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面将对本发明实施例中的技术方案进行清楚、完整地描述。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。
以下结合实施例对本发明的特征和性能作进一步的详细描述。
实施例1
1癫痫患者家系的FASN突变检测
1.1对象:来自重庆市江津区的黄氏癫痫家系先证者及其家系成员,该家系的遗传图谱如图1所示,检测对象包括2例癫痫患者(图中1 3号成员),1例正常家庭成员(图中2号成员);1号、3号:全面强直-阵挛发作;2号:未患病。对所有1-3号家庭成员进行详细体格检查,在签署知情同意书后每人采集血液样本。
1.2测序:
将采集的血液样本送北京迈基诺基因科技股份有限公司测序进行癫痫基因突变筛查及验证,分析临床表型、基因型的关系,结果如下:
将测得的序列与正常FASN标准序列比对(https://www.ncbi.nlm.nih.gov/nuccore/NM_004104.4),测序结果如图2-A所示,先证者即4号成员FASN有1个杂合突变:c.2998G>A(编码区第2998号核苷酸由鸟嘌呤突变为胸腺嘧啶),导致氨基酸改变p.D1000N(第1000位氨基酸由天冬氨基酸(D)变异为天冬酰胺(N)),为错义突变。该变异不属于多态性位点,在人群中发生频率极低。在HGMD专业版数据库中未见报道。经家系验证分析,在HGMD专业版数据库中未见报道。经家系验证分析,受检人(图1中3号成员)之父(图1中2号成员)该位点无变异,受检人之母(图1中1号成员)该位点杂合变异。图2的A-C中,箭头所示为FASN的2998位点的测序结果,1、3号成员在该位点的测序结果为均杂合突变(基因型为GT型),2号成员在该位点的测序结果为无突变(基因型为GG型)。
c.2998G>A变异信息如下:
DNA变异信息:(临床意义尚不明确,仅供参考)
综上分析可以看出,FASN基因的2998位点发生突G>A与癫痫发生相关,该位点可以作为辅助诊断癫痫疾病的靶点,且含有该突变c.G2998A的核酸片段或相应的具有突变p.D1000N的FASN蛋白均可以作为辅助诊断癫痫疾病的生物标志物。相应的,用于检测含有具有突变c.G2998A的核酸片段的探针、引物或其他试剂以及检测具有突变p.D1000N的FASN蛋白的抗体等均可以用于制备辅助诊断癫痫疾病的制剂盒。
实施例2
本实施例提供了检测FASN第2998位点突变的试剂盒,其包括:用于检测FASN2998位点是否发生突变G2998A的引物,正向引物:AAGCTGCATGCCTAGCTGTG,反向引物:GAACGGCAACCTGGTAGTGAG;以及PCR扩增试剂:10×Buffer(含15mM Mg2+)、dNTP(2.5Mm)、高保真DNA聚合酶pfu DNA聚合酶(5U/μl)以及ddH2O。
使用试剂盒的检测FASN第2998位点是否发生突变G>A的方法,主要包括如下步骤:
(1)提取样本DNA,以此为模板,利用上述PCR反应试剂盒进行PCR反应。
(2)多重PCR产物检测:用琼脂糖凝胶对PCR扩增产物进行电泳检测PCR是否成功扩增,扩增长度385bp。
(3)将PCR产物直接测序,将测序结果与正常FASN序列比对,可判断待测样本的FASN基因的第2998位点是否发生突变G>A。
以上所述仅为本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
SEQUENCE LISTING
<110> 重庆医科大学附属第一医院
<120> 一种辅助癫痫诊断的标志物及其检测试剂盒
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 7536
<212> DNA
<213> 人工序列
<400> 1
atggaggagg tggtgattgc cggcatgtcc gggaagctgc cagagtcgga gaacttgcag 60
gagttctggg acaacctcat cggcggtgtg gacatggtca cggacgatga ccgtcgctgg 120
aaggcggggc tctacggcct gccccggcgg tccggcaagc tgaaggacct gtctaggttt 180
gatgcctcct tcttcggagt ccaccccaag caggcacaca cgatggaccc tcagctgcgg 240
ctgctgctgg aagtcaccta tgaagccatc gtggacggag gcatcaaccc agattcactc 300
cgaggaacac acactggcgt ctgggtgggc gtgagcggct ctgagacctc ggaggccctg 360
agccgagacc ccgagacact cgtgggctac agcatggtgg gctgccagcg agcgatgatg 420
gccaaccggc tctccttctt cttcgacttc agagggccca gcatcgcact ggacacagcc 480
tgctcctcca gcctgatggc cctgcagaac gcctaccagg ccatccacag cgggcagtgc 540
cctgccgcca tcgtgggggg catcaatgtc ctgctgaagc ccaacacctc cgtgcagttc 600
ttgaggctgg ggatgctcag ccccgagggc acctgcaagg ccttcgacac agcggggaat 660
gggtactgcc gctcggaggg tgtggtggcc gtcctgctga ccaagaagtc cctggcccgg 720
cgggtgtacg ccaccatcct gaacgccggc accaatacag atggcttcaa ggagcaaggc 780
gtgaccttcc cctcagggga tatccaggag cagctcatcc gctcgttgta ccagtcggcc 840
ggagtggccc ctgagtcatt tgaatacatc gaagcccacg gcacaggcac caaggtgggc 900
gacccccagg agctgaatgg catcacccga gccctgtgcg ccacccgcca ggagccgctg 960
ctcatcggct ccaccaagtc caacatgggg cacccggagc cagcctcggg gctggcagcc 1020
ctggccaagg tgctgctgtc cctggagcac gggctctggg cccccaacct gcacttccat 1080
agccccaacc ctgagatccc agcgctgttg gatgggcggc tgcaggtggt ggaccagccc 1140
ctgcccgtcc gtggcggcaa cgtgggcatc aactcctttg gcttcggggg ctccaacgtg 1200
cacatcatcc tgaggcccaa cacgcagccg ccccccgcac ccgccccaca tgccaccctg 1260
ccccgtctgc tgcgggccag cggacgcacc cctgaggccg tgcagaagct gctggagcag 1320
ggcctccggc acagccagga cctggctttc ctgagcatgc tgaacgacat cgcggctgtc 1380
cccgccaccg ccatgccctt ccgtggctac gctgtgctgg gtggtgagcg cggtggccca 1440
gaggtgcagc aggtgcccgc tggcgagcgc ccgctctggt tcatctgctc tgggatgggc 1500
acacagtggc gcgggatggg gctgagcctc atgcgcctgg accgcttccg agattccatc 1560
ctacgctccg atgaggctgt gaagccattc ggcctgaagg tgtcacagct gctgctgagc 1620
acagacgaga gcacctttga tgacatcgtc cattcgtttg tgagcctgac tgccatccag 1680
ataggcctca tagacctgct gagctgcatg gggctgaggc cagatggcat cgtcggccac 1740
tccctggggg aggtggcctg tggctacgcc gacggctgcc tgtcccagga ggaggccgtc 1800
ctcgctgcct actggagggg acagtgcatc aaagaagccc atctcccgcc gggcgccatg 1860
gcagccgtgg gcttgtcctg ggaggagtgt aaacagcgct gccccccggg cgtggtgccc 1920
gcctgccaca actccaagga cacagtcacc atctcgggac ctcaggcccc ggtgtttgag 1980
ttcgtggagc agctgaggaa ggagggtgtg tttgccaagg aggtgcggac cggcggtatg 2040
gccttccact cctacttcat ggaggccatc gcacccccac tgctgcagga gctcaagaag 2100
gtgatccggg agccgaagcc acgttcagcc cgctggctca gcacctctat ccccgaggcc 2160
cagtggcaca gcagcctggc acgcacgtcc tccgccgagt acaatgtcaa caacctggtg 2220
agccctgtgc tgttccagga ggccctgtgg cacgtgcctg agcacgcggt ggtgctggag 2280
atcgcgcccc acgccctgct gcaggctgtc ctgaagcgtg gcctgaagcc gagctgcacc 2340
atcatccccc tgatgaagaa ggatcacagg gacaacctgg agttcttcct ggccggcatc 2400
ggcaggctgc acctctcagg catcgacgcc aaccccaatg ccttgttccc acctgtggag 2460
ttcccagctc cccgaggaac tcccctcatc tccccactca tcaagtggga ccacagcctg 2520
gcctgggacg tgccggccgc cgaggacttc cccaacggtt caggttcccc ctcagccgcc 2580
atctacaaca tcgacaccag ctccgagtct cctgaccact acctggtgga ccacaccctc 2640
gacggtcgcg tcctcttccc cgccactggc tacctgagca tagtgtggaa gacgctggcc 2700
cgcgccctgg gcctgggcgt cgagcagctg cctgtggtgt ttgaggatgt ggtgctgcac 2760
caggccacca tcctgcccaa gactgggaca gtgtccctgg aggtacggct cctggaggcc 2820
tcccgtgcct tcgaggtgtc agagaacggc aacctggtag tgagtgggaa ggtgtaccag 2880
tgggatgacc ctgaccccag gctcttcgac cacccggaaa gccccacccc caaccccacg 2940
gagcccctct tcctggccca ggctgaagtt tacaaggagc tgcgtctgcg tggctacaac 3000
tacggccctc atttccaggg catcctggag gccagcctgg aaggtgactc ggggaggctg 3060
ctgtggaagg ataactgggt gagcttcatg gacaccatgc tgcagatgtc catcctgggc 3120
tcggccaagc acggcctgta cctgcccacc cgtgtcaccg ccatccacat cgaccctgcc 3180
acccacaggc agaagctgta cacactgcag gacaaggccc aagtggctga cgtggtggtg 3240
agcaggtggc tgagggtcac agtggccgga ggcgtccaca tctccgggct ccacactgag 3300
tcggccccgc ggcggcagca ggagcagcag gtgcccatcc tggagaagtt ttgcttcact 3360
ccccacacgg aggaggggtg cctgtctgag cgcgctgccc tgcaggagga gctgcaactg 3420
tgcaaggggc tggtgcaggc actgcagacc aaggtgaccc agcaggggct gaagatggtg 3480
gtgcccggac tggatggggc ccagatcccc cgggacccct cacagcagga actgccccgg 3540
ctgttgtcgg ctgcctgcag gcttcagctc aacgggaacc tgcagctgga gctggcgcag 3600
gtgctggccc aggagaggcc caagctgcca gaggaccctc tgctcagcgg cctcctggac 3660
tccccggcac tcaaggcctg cctggacact gccgtggaga acatgcccag cctgaagatg 3720
aaggtggtgg aggtgctggc tggccacggt cacctgtatt cccgcatccc aggcctgctc 3780
agcccccatc ccctgctgca gctgagctac acggccaccg accgccaccc ccaggccctg 3840
gaggctgccc aggccgagct gcagcagcac gacgttgccc agggccagtg ggatcccgca 3900
gaccctgccc ccagcgccct gggcagcgcc gacctcctgg tgtgcaactg tgctgtggct 3960
gccctcgggg acccggcctc agctctcagc aacatggtgg ctgccctgag agaagggggc 4020
tttctgctcc tgcacacact gctccggggg caccccctcg gggacatcgt ggccttcctc 4080
acctccactg agccgcagta tggccagggc atcctgagcc aggacgcgtg ggagagcctc 4140
ttctccaggg tgtcgctgcg cctggtgggc ctgaagaagt ccttctacgg ctccacgctc 4200
ttcctgtgcc gccggcccac cccgcaggac agccccatct tcctgccggt ggacgatacc 4260
agcttccgct gggtggagtc tctgaagggc atcctggctg acgaagactc ttcccggcct 4320
gtgtggctga aggccatcaa ctgtgccacc tcgggcgtgg tgggcttggt gaactgtctc 4380
cgccgagagc ccggcgggaa ccgcctccgg tgtgtgctgc tctccaacct cagcagcacc 4440
tcccacgtcc cggaggtgga cccgggctcc gcagaactgc agaaggtgtt gcagggagac 4500
ctggtgatga acgtctaccg cgacggggcc tggggggctt tccgccactt cctgctggag 4560
gaggacaagc ctgaggagcc gacggcacat gcctttgtga gcaccctcac ccggggggac 4620
ctgtcctcca tccgctgggt ctgctcctcg ctgcgccatg cccagcccac ctgccctggc 4680
gcccagctct gcacggtcta ctacgcctcc ctcaacttcc gcgacatcat gctggccact 4740
ggcaagctgt cccctgatgc catcccaggg aagtggacct cccaggacag cctgctaggt 4800
atggagttct cgggccgaga cgccagcggc aagcgtgtga tgggactggt gcctgccaag 4860
ggcctggcca cctctgtcct gctgtcaccg gacttcctct gggatgtgcc ttccaactgg 4920
acgctggagg aggcggcctc ggtgcctgtc gtctacagca cggcctacta cgcgctggtg 4980
gtgcgtgggc gggtgcgccc cggggagacg ctgctcatcc actcgggctc gggcggcgtg 5040
ggccaggccg ccatcgccat cgccctcagt ctgggctgcc gcgtcttcac caccgtgggg 5100
tcggctgaga agcgggcgta cctccaggcc aggttccccc agctcgacag caccagcttc 5160
gccaactccc gggacacatc cttcgagcag catgtgctgt ggcacacggg cgggaagggc 5220
gttgacctgg tcttgaactc cttggcggaa gagaagctgc aggccagcgt gaggtgcttg 5280
gctacgcacg gtcgcttcct ggaaattggc aaattcgacc tttctcagaa ccacccgctc 5340
ggcatggcta tcttcctgaa gaacgtgaca ttccacgggg tcctactgga tgcgttcttc 5400
aacgagagca gtgctgactg gcgggaggtg tgggcgcttg tgcaggccgg catccgggat 5460
ggggtggtac ggcccctcaa gtgcacggtg ttccatgggg cccaggtgga ggacgccttc 5520
cgctacatgg cccaagggaa gcacattggc aaagtcgtcg tgcaggtgct tgcggaggag 5580
ccggaggcag tgctgaaggg ggccaaaccc aagctgatgt cggccatctc caagaccttc 5640
tgcccggccc acaagagcta catcatcgct ggtggtctgg gtggcttcgg cctggagttg 5700
gcgcagtggc tgatacagcg tggggtgcag aagctcgtgt tgacttctcg ctccgggatc 5760
cggacaggct accaggccaa gcaggtccgc cggtggaggc gccagggcgt acaggtgcag 5820
gtgtccacca gcaacatcag ctcactggag ggggcccggg gcctcattgc cgaggcggcg 5880
cagcttgggc ccgtgggcgg cgtcttcaac ctggccgtgg tcttgagaga tggcttgctg 5940
gagaaccaga ccccagagtt cttccaggac gtctgcaagc ccaagtacag cggcaccctg 6000
aacctggaca gggtgacccg agaggcgtgc cctgagctgg actactttgt ggtcttctcc 6060
tctgtgagct gcgggcgtgg caatgcggga cagagcaact acggctttgc caattccgcc 6120
atggagcgta tctgtgagaa acgccggcac gaaggcctcc caggcctggc cgtgcagtgg 6180
ggcgccatcg gcgacgtggg cattttggtg gagacgatga gcaccaacga cacgatcgtc 6240
agtggcacgc tgccccagcg catggcgtcc tgcctggagg tgctggacct cttcctgaac 6300
cagccccaca tggtcctgag cagctttgtg ctggctgaga aggctgcggc ctatagggac 6360
agggacagcc agcgggacct ggtggaggcc gtggcacaca tcctgggcat ccgcgacttg 6420
gctgctgtca acctggacag ctcactggcg gacctgggcc tggactcgct catgagcgtg 6480
gaggtgcgcc agacgctgga gcgtgagctc aacctggtgc tgtccgtgcg cgaggtgcgg 6540
caactcacgc tccggaaact gcaggagctg tcctcaaagg cggatgaggc cagcgagctg 6600
gcatgcccca cgcccaagga ggatggtctg gcccagcagc agactcagct gaacctgcgc 6660
tccctgctgg tgaacccgga gggccccacc ctgatgcggc tcaactccgt gcagagctcg 6720
gagcggcccc tgttcctggt gcacccaatc gagggctcca ccaccgtgtt ccacagcctg 6780
gcctcccggc tcagcatccc cacctatggc ctgcagtgca cccgagctgc gccccttgac 6840
agcatccaca gcctggctgc ctactacatc gactgcatca ggcaggtgca gcccgagggc 6900
ccctaccgcg tggccggcta ctcctacggg gcctgcgtgg cctttgaaat gtgctcccag 6960
ctgcaggccc agcagagccc agcccccacc cacaacagcc tcttcctgtt cgacggctcg 7020
cccacctacg tactggccta cacccagagc taccgggcaa agctgacccc aggctgtgag 7080
gctgaggctg agacggaggc catatgcttc ttcgtgcagc agttcacgga catggagcac 7140
aacagggtgc tggaggcgct gctgccgctg aagggcctag aggagcgtgt ggcagccgcc 7200
gtggacctga tcatcaagag ccaccagggc ctggaccgcc aggagctgag ctttgcggcc 7260
cggtccttct actacaagct gcgtgccgct gagcagtaca cacccaaggc caagtaccat 7320
ggcaacgtga tgctactgcg cgccaagacg ggtggcgcct acggcgagga cctgggcgcg 7380
gactacaacc tctcccaggt atgcgacggg aaagtatccg tccacgtcat cgagggtgac 7440
caccgcacgc tgctggaggg cagcggcctg gagtccatca tcagcatcat ccacagctcc 7500
ctggctgagc cacgcgtgag cgtgcgggag ggctag 7536
<210> 2
<211> 2511
<212> PRT
<213> 人工序列
<400> 2
Met Glu Glu Val Val Ile Ala Gly Met Ser Gly Lys Leu Pro Glu Ser
1 5 10 15
Glu Asn Leu Gln Glu Phe Trp Asp Asn Leu Ile Gly Gly Val Asp Met
20 25 30
Val Thr Asp Asp Asp Arg Arg Trp Lys Ala Gly Leu Tyr Gly Leu Pro
35 40 45
Arg Arg Ser Gly Lys Leu Lys Asp Leu Ser Arg Phe Asp Ala Ser Phe
50 55 60
Phe Gly Val His Pro Lys Gln Ala His Thr Met Asp Pro Gln Leu Arg
65 70 75 80
Leu Leu Leu Glu Val Thr Tyr Glu Ala Ile Val Asp Gly Gly Ile Asn
85 90 95
Pro Asp Ser Leu Arg Gly Thr His Thr Gly Val Trp Val Gly Val Ser
100 105 110
Gly Ser Glu Thr Ser Glu Ala Leu Ser Arg Asp Pro Glu Thr Leu Val
115 120 125
Gly Tyr Ser Met Val Gly Cys Gln Arg Ala Met Met Ala Asn Arg Leu
130 135 140
Ser Phe Phe Phe Asp Phe Arg Gly Pro Ser Ile Ala Leu Asp Thr Ala
145 150 155 160
Cys Ser Ser Ser Leu Met Ala Leu Gln Asn Ala Tyr Gln Ala Ile His
165 170 175
Ser Gly Gln Cys Pro Ala Ala Ile Val Gly Gly Ile Asn Val Leu Leu
180 185 190
Lys Pro Asn Thr Ser Val Gln Phe Leu Arg Leu Gly Met Leu Ser Pro
195 200 205
Glu Gly Thr Cys Lys Ala Phe Asp Thr Ala Gly Asn Gly Tyr Cys Arg
210 215 220
Ser Glu Gly Val Val Ala Val Leu Leu Thr Lys Lys Ser Leu Ala Arg
225 230 235 240
Arg Val Tyr Ala Thr Ile Leu Asn Ala Gly Thr Asn Thr Asp Gly Phe
245 250 255
Lys Glu Gln Gly Val Thr Phe Pro Ser Gly Asp Ile Gln Glu Gln Leu
260 265 270
Ile Arg Ser Leu Tyr Gln Ser Ala Gly Val Ala Pro Glu Ser Phe Glu
275 280 285
Tyr Ile Glu Ala His Gly Thr Gly Thr Lys Val Gly Asp Pro Gln Glu
290 295 300
Leu Asn Gly Ile Thr Arg Ala Leu Cys Ala Thr Arg Gln Glu Pro Leu
305 310 315 320
Leu Ile Gly Ser Thr Lys Ser Asn Met Gly His Pro Glu Pro Ala Ser
325 330 335
Gly Leu Ala Ala Leu Ala Lys Val Leu Leu Ser Leu Glu His Gly Leu
340 345 350
Trp Ala Pro Asn Leu His Phe His Ser Pro Asn Pro Glu Ile Pro Ala
355 360 365
Leu Leu Asp Gly Arg Leu Gln Val Val Asp Gln Pro Leu Pro Val Arg
370 375 380
Gly Gly Asn Val Gly Ile Asn Ser Phe Gly Phe Gly Gly Ser Asn Val
385 390 395 400
His Ile Ile Leu Arg Pro Asn Thr Gln Pro Pro Pro Ala Pro Ala Pro
405 410 415
His Ala Thr Leu Pro Arg Leu Leu Arg Ala Ser Gly Arg Thr Pro Glu
420 425 430
Ala Val Gln Lys Leu Leu Glu Gln Gly Leu Arg His Ser Gln Asp Leu
435 440 445
Ala Phe Leu Ser Met Leu Asn Asp Ile Ala Ala Val Pro Ala Thr Ala
450 455 460
Met Pro Phe Arg Gly Tyr Ala Val Leu Gly Gly Glu Arg Gly Gly Pro
465 470 475 480
Glu Val Gln Gln Val Pro Ala Gly Glu Arg Pro Leu Trp Phe Ile Cys
485 490 495
Ser Gly Met Gly Thr Gln Trp Arg Gly Met Gly Leu Ser Leu Met Arg
500 505 510
Leu Asp Arg Phe Arg Asp Ser Ile Leu Arg Ser Asp Glu Ala Val Lys
515 520 525
Pro Phe Gly Leu Lys Val Ser Gln Leu Leu Leu Ser Thr Asp Glu Ser
530 535 540
Thr Phe Asp Asp Ile Val His Ser Phe Val Ser Leu Thr Ala Ile Gln
545 550 555 560
Ile Gly Leu Ile Asp Leu Leu Ser Cys Met Gly Leu Arg Pro Asp Gly
565 570 575
Ile Val Gly His Ser Leu Gly Glu Val Ala Cys Gly Tyr Ala Asp Gly
580 585 590
Cys Leu Ser Gln Glu Glu Ala Val Leu Ala Ala Tyr Trp Arg Gly Gln
595 600 605
Cys Ile Lys Glu Ala His Leu Pro Pro Gly Ala Met Ala Ala Val Gly
610 615 620
Leu Ser Trp Glu Glu Cys Lys Gln Arg Cys Pro Pro Gly Val Val Pro
625 630 635 640
Ala Cys His Asn Ser Lys Asp Thr Val Thr Ile Ser Gly Pro Gln Ala
645 650 655
Pro Val Phe Glu Phe Val Glu Gln Leu Arg Lys Glu Gly Val Phe Ala
660 665 670
Lys Glu Val Arg Thr Gly Gly Met Ala Phe His Ser Tyr Phe Met Glu
675 680 685
Ala Ile Ala Pro Pro Leu Leu Gln Glu Leu Lys Lys Val Ile Arg Glu
690 695 700
Pro Lys Pro Arg Ser Ala Arg Trp Leu Ser Thr Ser Ile Pro Glu Ala
705 710 715 720
Gln Trp His Ser Ser Leu Ala Arg Thr Ser Ser Ala Glu Tyr Asn Val
725 730 735
Asn Asn Leu Val Ser Pro Val Leu Phe Gln Glu Ala Leu Trp His Val
740 745 750
Pro Glu His Ala Val Val Leu Glu Ile Ala Pro His Ala Leu Leu Gln
755 760 765
Ala Val Leu Lys Arg Gly Leu Lys Pro Ser Cys Thr Ile Ile Pro Leu
770 775 780
Met Lys Lys Asp His Arg Asp Asn Leu Glu Phe Phe Leu Ala Gly Ile
785 790 795 800
Gly Arg Leu His Leu Ser Gly Ile Asp Ala Asn Pro Asn Ala Leu Phe
805 810 815
Pro Pro Val Glu Phe Pro Ala Pro Arg Gly Thr Pro Leu Ile Ser Pro
820 825 830
Leu Ile Lys Trp Asp His Ser Leu Ala Trp Asp Val Pro Ala Ala Glu
835 840 845
Asp Phe Pro Asn Gly Ser Gly Ser Pro Ser Ala Ala Ile Tyr Asn Ile
850 855 860
Asp Thr Ser Ser Glu Ser Pro Asp His Tyr Leu Val Asp His Thr Leu
865 870 875 880
Asp Gly Arg Val Leu Phe Pro Ala Thr Gly Tyr Leu Ser Ile Val Trp
885 890 895
Lys Thr Leu Ala Arg Ala Leu Gly Leu Gly Val Glu Gln Leu Pro Val
900 905 910
Val Phe Glu Asp Val Val Leu His Gln Ala Thr Ile Leu Pro Lys Thr
915 920 925
Gly Thr Val Ser Leu Glu Val Arg Leu Leu Glu Ala Ser Arg Ala Phe
930 935 940
Glu Val Ser Glu Asn Gly Asn Leu Val Val Ser Gly Lys Val Tyr Gln
945 950 955 960
Trp Asp Asp Pro Asp Pro Arg Leu Phe Asp His Pro Glu Ser Pro Thr
965 970 975
Pro Asn Pro Thr Glu Pro Leu Phe Leu Ala Gln Ala Glu Val Tyr Lys
980 985 990
Glu Leu Arg Leu Arg Gly Tyr Asn Tyr Gly Pro His Phe Gln Gly Ile
995 1000 1005
Leu Glu Ala Ser Leu Glu Gly Asp Ser Gly Arg Leu Leu Trp Lys
1010 1015 1020
Asp Asn Trp Val Ser Phe Met Asp Thr Met Leu Gln Met Ser Ile
1025 1030 1035
Leu Gly Ser Ala Lys His Gly Leu Tyr Leu Pro Thr Arg Val Thr
1040 1045 1050
Ala Ile His Ile Asp Pro Ala Thr His Arg Gln Lys Leu Tyr Thr
1055 1060 1065
Leu Gln Asp Lys Ala Gln Val Ala Asp Val Val Val Ser Arg Trp
1070 1075 1080
Leu Arg Val Thr Val Ala Gly Gly Val His Ile Ser Gly Leu His
1085 1090 1095
Thr Glu Ser Ala Pro Arg Arg Gln Gln Glu Gln Gln Val Pro Ile
1100 1105 1110
Leu Glu Lys Phe Cys Phe Thr Pro His Thr Glu Glu Gly Cys Leu
1115 1120 1125
Ser Glu Arg Ala Ala Leu Gln Glu Glu Leu Gln Leu Cys Lys Gly
1130 1135 1140
Leu Val Gln Ala Leu Gln Thr Lys Val Thr Gln Gln Gly Leu Lys
1145 1150 1155
Met Val Val Pro Gly Leu Asp Gly Ala Gln Ile Pro Arg Asp Pro
1160 1165 1170
Ser Gln Gln Glu Leu Pro Arg Leu Leu Ser Ala Ala Cys Arg Leu
1175 1180 1185
Gln Leu Asn Gly Asn Leu Gln Leu Glu Leu Ala Gln Val Leu Ala
1190 1195 1200
Gln Glu Arg Pro Lys Leu Pro Glu Asp Pro Leu Leu Ser Gly Leu
1205 1210 1215
Leu Asp Ser Pro Ala Leu Lys Ala Cys Leu Asp Thr Ala Val Glu
1220 1225 1230
Asn Met Pro Ser Leu Lys Met Lys Val Val Glu Val Leu Ala Gly
1235 1240 1245
His Gly His Leu Tyr Ser Arg Ile Pro Gly Leu Leu Ser Pro His
1250 1255 1260
Pro Leu Leu Gln Leu Ser Tyr Thr Ala Thr Asp Arg His Pro Gln
1265 1270 1275
Ala Leu Glu Ala Ala Gln Ala Glu Leu Gln Gln His Asp Val Ala
1280 1285 1290
Gln Gly Gln Trp Asp Pro Ala Asp Pro Ala Pro Ser Ala Leu Gly
1295 1300 1305
Ser Ala Asp Leu Leu Val Cys Asn Cys Ala Val Ala Ala Leu Gly
1310 1315 1320
Asp Pro Ala Ser Ala Leu Ser Asn Met Val Ala Ala Leu Arg Glu
1325 1330 1335
Gly Gly Phe Leu Leu Leu His Thr Leu Leu Arg Gly His Pro Leu
1340 1345 1350
Gly Asp Ile Val Ala Phe Leu Thr Ser Thr Glu Pro Gln Tyr Gly
1355 1360 1365
Gln Gly Ile Leu Ser Gln Asp Ala Trp Glu Ser Leu Phe Ser Arg
1370 1375 1380
Val Ser Leu Arg Leu Val Gly Leu Lys Lys Ser Phe Tyr Gly Ser
1385 1390 1395
Thr Leu Phe Leu Cys Arg Arg Pro Thr Pro Gln Asp Ser Pro Ile
1400 1405 1410
Phe Leu Pro Val Asp Asp Thr Ser Phe Arg Trp Val Glu Ser Leu
1415 1420 1425
Lys Gly Ile Leu Ala Asp Glu Asp Ser Ser Arg Pro Val Trp Leu
1430 1435 1440
Lys Ala Ile Asn Cys Ala Thr Ser Gly Val Val Gly Leu Val Asn
1445 1450 1455
Cys Leu Arg Arg Glu Pro Gly Gly Asn Arg Leu Arg Cys Val Leu
1460 1465 1470
Leu Ser Asn Leu Ser Ser Thr Ser His Val Pro Glu Val Asp Pro
1475 1480 1485
Gly Ser Ala Glu Leu Gln Lys Val Leu Gln Gly Asp Leu Val Met
1490 1495 1500
Asn Val Tyr Arg Asp Gly Ala Trp Gly Ala Phe Arg His Phe Leu
1505 1510 1515
Leu Glu Glu Asp Lys Pro Glu Glu Pro Thr Ala His Ala Phe Val
1520 1525 1530
Ser Thr Leu Thr Arg Gly Asp Leu Ser Ser Ile Arg Trp Val Cys
1535 1540 1545
Ser Ser Leu Arg His Ala Gln Pro Thr Cys Pro Gly Ala Gln Leu
1550 1555 1560
Cys Thr Val Tyr Tyr Ala Ser Leu Asn Phe Arg Asp Ile Met Leu
1565 1570 1575
Ala Thr Gly Lys Leu Ser Pro Asp Ala Ile Pro Gly Lys Trp Thr
1580 1585 1590
Ser Gln Asp Ser Leu Leu Gly Met Glu Phe Ser Gly Arg Asp Ala
1595 1600 1605
Ser Gly Lys Arg Val Met Gly Leu Val Pro Ala Lys Gly Leu Ala
1610 1615 1620
Thr Ser Val Leu Leu Ser Pro Asp Phe Leu Trp Asp Val Pro Ser
1625 1630 1635
Asn Trp Thr Leu Glu Glu Ala Ala Ser Val Pro Val Val Tyr Ser
1640 1645 1650
Thr Ala Tyr Tyr Ala Leu Val Val Arg Gly Arg Val Arg Pro Gly
1655 1660 1665
Glu Thr Leu Leu Ile His Ser Gly Ser Gly Gly Val Gly Gln Ala
1670 1675 1680
Ala Ile Ala Ile Ala Leu Ser Leu Gly Cys Arg Val Phe Thr Thr
1685 1690 1695
Val Gly Ser Ala Glu Lys Arg Ala Tyr Leu Gln Ala Arg Phe Pro
1700 1705 1710
Gln Leu Asp Ser Thr Ser Phe Ala Asn Ser Arg Asp Thr Ser Phe
1715 1720 1725
Glu Gln His Val Leu Trp His Thr Gly Gly Lys Gly Val Asp Leu
1730 1735 1740
Val Leu Asn Ser Leu Ala Glu Glu Lys Leu Gln Ala Ser Val Arg
1745 1750 1755
Cys Leu Ala Thr His Gly Arg Phe Leu Glu Ile Gly Lys Phe Asp
1760 1765 1770
Leu Ser Gln Asn His Pro Leu Gly Met Ala Ile Phe Leu Lys Asn
1775 1780 1785
Val Thr Phe His Gly Val Leu Leu Asp Ala Phe Phe Asn Glu Ser
1790 1795 1800
Ser Ala Asp Trp Arg Glu Val Trp Ala Leu Val Gln Ala Gly Ile
1805 1810 1815
Arg Asp Gly Val Val Arg Pro Leu Lys Cys Thr Val Phe His Gly
1820 1825 1830
Ala Gln Val Glu Asp Ala Phe Arg Tyr Met Ala Gln Gly Lys His
1835 1840 1845
Ile Gly Lys Val Val Val Gln Val Leu Ala Glu Glu Pro Glu Ala
1850 1855 1860
Val Leu Lys Gly Ala Lys Pro Lys Leu Met Ser Ala Ile Ser Lys
1865 1870 1875
Thr Phe Cys Pro Ala His Lys Ser Tyr Ile Ile Ala Gly Gly Leu
1880 1885 1890
Gly Gly Phe Gly Leu Glu Leu Ala Gln Trp Leu Ile Gln Arg Gly
1895 1900 1905
Val Gln Lys Leu Val Leu Thr Ser Arg Ser Gly Ile Arg Thr Gly
1910 1915 1920
Tyr Gln Ala Lys Gln Val Arg Arg Trp Arg Arg Gln Gly Val Gln
1925 1930 1935
Val Gln Val Ser Thr Ser Asn Ile Ser Ser Leu Glu Gly Ala Arg
1940 1945 1950
Gly Leu Ile Ala Glu Ala Ala Gln Leu Gly Pro Val Gly Gly Val
1955 1960 1965
Phe Asn Leu Ala Val Val Leu Arg Asp Gly Leu Leu Glu Asn Gln
1970 1975 1980
Thr Pro Glu Phe Phe Gln Asp Val Cys Lys Pro Lys Tyr Ser Gly
1985 1990 1995
Thr Leu Asn Leu Asp Arg Val Thr Arg Glu Ala Cys Pro Glu Leu
2000 2005 2010
Asp Tyr Phe Val Val Phe Ser Ser Val Ser Cys Gly Arg Gly Asn
2015 2020 2025
Ala Gly Gln Ser Asn Tyr Gly Phe Ala Asn Ser Ala Met Glu Arg
2030 2035 2040
Ile Cys Glu Lys Arg Arg His Glu Gly Leu Pro Gly Leu Ala Val
2045 2050 2055
Gln Trp Gly Ala Ile Gly Asp Val Gly Ile Leu Val Glu Thr Met
2060 2065 2070
Ser Thr Asn Asp Thr Ile Val Ser Gly Thr Leu Pro Gln Arg Met
2075 2080 2085
Ala Ser Cys Leu Glu Val Leu Asp Leu Phe Leu Asn Gln Pro His
2090 2095 2100
Met Val Leu Ser Ser Phe Val Leu Ala Glu Lys Ala Ala Ala Tyr
2105 2110 2115
Arg Asp Arg Asp Ser Gln Arg Asp Leu Val Glu Ala Val Ala His
2120 2125 2130
Ile Leu Gly Ile Arg Asp Leu Ala Ala Val Asn Leu Asp Ser Ser
2135 2140 2145
Leu Ala Asp Leu Gly Leu Asp Ser Leu Met Ser Val Glu Val Arg
2150 2155 2160
Gln Thr Leu Glu Arg Glu Leu Asn Leu Val Leu Ser Val Arg Glu
2165 2170 2175
Val Arg Gln Leu Thr Leu Arg Lys Leu Gln Glu Leu Ser Ser Lys
2180 2185 2190
Ala Asp Glu Ala Ser Glu Leu Ala Cys Pro Thr Pro Lys Glu Asp
2195 2200 2205
Gly Leu Ala Gln Gln Gln Thr Gln Leu Asn Leu Arg Ser Leu Leu
2210 2215 2220
Val Asn Pro Glu Gly Pro Thr Leu Met Arg Leu Asn Ser Val Gln
2225 2230 2235
Ser Ser Glu Arg Pro Leu Phe Leu Val His Pro Ile Glu Gly Ser
2240 2245 2250
Thr Thr Val Phe His Ser Leu Ala Ser Arg Leu Ser Ile Pro Thr
2255 2260 2265
Tyr Gly Leu Gln Cys Thr Arg Ala Ala Pro Leu Asp Ser Ile His
2270 2275 2280
Ser Leu Ala Ala Tyr Tyr Ile Asp Cys Ile Arg Gln Val Gln Pro
2285 2290 2295
Glu Gly Pro Tyr Arg Val Ala Gly Tyr Ser Tyr Gly Ala Cys Val
2300 2305 2310
Ala Phe Glu Met Cys Ser Gln Leu Gln Ala Gln Gln Ser Pro Ala
2315 2320 2325
Pro Thr His Asn Ser Leu Phe Leu Phe Asp Gly Ser Pro Thr Tyr
2330 2335 2340
Val Leu Ala Tyr Thr Gln Ser Tyr Arg Ala Lys Leu Thr Pro Gly
2345 2350 2355
Cys Glu Ala Glu Ala Glu Thr Glu Ala Ile Cys Phe Phe Val Gln
2360 2365 2370
Gln Phe Thr Asp Met Glu His Asn Arg Val Leu Glu Ala Leu Leu
2375 2380 2385
Pro Leu Lys Gly Leu Glu Glu Arg Val Ala Ala Ala Val Asp Leu
2390 2395 2400
Ile Ile Lys Ser His Gln Gly Leu Asp Arg Gln Glu Leu Ser Phe
2405 2410 2415
Ala Ala Arg Ser Phe Tyr Tyr Lys Leu Arg Ala Ala Glu Gln Tyr
2420 2425 2430
Thr Pro Lys Ala Lys Tyr His Gly Asn Val Met Leu Leu Arg Ala
2435 2440 2445
Lys Thr Gly Gly Ala Tyr Gly Glu Asp Leu Gly Ala Asp Tyr Asn
2450 2455 2460
Leu Ser Gln Val Cys Asp Gly Lys Val Ser Val His Val Ile Glu
2465 2470 2475
Gly Asp His Arg Thr Leu Leu Glu Gly Ser Gly Leu Glu Ser Ile
2480 2485 2490
Ile Ser Ile Ile His Ser Ser Leu Ala Glu Pro Arg Val Ser Val
2495 2500 2505
Arg Glu Gly
2510
Claims (2)
1.检测生物标志物的引物或探针在制备癫痫辅助诊断试剂盒、癫痫产前诊断试剂盒或新生儿癫痫筛查试剂盒中的应用,其特征在于,所述生物标志物为核酸分子,其相对于正常FASN的编码序列具有突变c.G2998A,所述正常FASN的转录本编号为NM_004104,所述标志物的碱基序列如SEQ ID NO.1所示;或者所述标志物为蛋白,其相对于正常FASN蛋白具有突变p.D1000N,所述标志物的氨基酸序列如SEQ ID NO.2所示,所述正常FASN蛋白由所述正常FASN编码。
2.根据权利要求1所述的应用,其特征在于,所述探针相对于所述正常FASN的编码序列的第2998位的碱基为A。
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