CN112831474A - 一株宽裂解谱沙门氏菌噬菌体rdp-nsa-19050及其应用 - Google Patents
一株宽裂解谱沙门氏菌噬菌体rdp-nsa-19050及其应用 Download PDFInfo
- Publication number
- CN112831474A CN112831474A CN202110053070.3A CN202110053070A CN112831474A CN 112831474 A CN112831474 A CN 112831474A CN 202110053070 A CN202110053070 A CN 202110053070A CN 112831474 A CN112831474 A CN 112831474A
- Authority
- CN
- China
- Prior art keywords
- salmonella
- phage
- nsa
- rdp
- bacteriophage
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/40—Viruses, e.g. bacteriophages
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/195—Antibiotics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/76—Viruses; Subviral particles; Bacteriophages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/16—Organic compounds
- C11D3/38—Products with no well-defined composition, e.g. natural products
- C11D3/381—Microorganisms
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11D—DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C11D3/00—Other compounding ingredients of detergent compositions covered in group C11D1/00
- C11D3/48—Medical, disinfecting agents, disinfecting, antibacterial, germicidal or antimicrobial compositions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10111—Myoviridae
- C12N2795/10121—Viruses as such, e.g. new isolates, mutants or their genomic sequences
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10111—Myoviridae
- C12N2795/10131—Uses of virus other than therapeutic or vaccine, e.g. disinfectant
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2795/00—Bacteriophages
- C12N2795/00011—Details
- C12N2795/10011—Details dsDNA Bacteriophages
- C12N2795/10111—Myoviridae
- C12N2795/10132—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Polymers & Plastics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Animal Husbandry (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Biochemistry (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- General Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Environmental Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Mycology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
Abstract
本发明公开了一株宽裂解谱沙门氏菌噬菌体RDP‑NSA‑19050,保藏编号为CGMCCNo.21415,基因长度87591bp,对全基因进行测序后发现,不具有毒力基因和溶原性基因,本发明提供的沙门氏菌噬菌体RDP‑NSA‑19050可以裂解包括斯坦利沙门氏菌、鸡伤寒沙门氏菌、鼠伤寒沙门氏菌、甲型副伤寒沙门氏菌、乙型副伤寒沙门氏菌、伤寒沙门氏菌、肠炎沙门氏菌、肯坦基沙门氏菌在内的多种致病菌,具有裂解谱广、裂解率高的特点,该噬菌体既可以单独使用,也可以和其他大肠杆菌、沙门氏菌的噬菌体和益生菌复配使用,可以有效降低感染沙门氏菌雏鸡的死亡率。
Description
技术领域
本发明涉及生物技术领域,特别是涉及一株宽裂解谱沙门氏菌噬菌体RDP-NSA-19050及其在禽类动物致病性沙门氏菌感染中的应用。
背景技术
沙门氏菌病是公共卫生学上具有重要意义的人畜共患病之一,是一种常见的食源性致病菌。鸡是沙门氏菌最重要的宿主,沙门氏菌可引起鸡急性或慢性疾病,感染沙门氏菌的鸡也可通过产出带菌的蛋导致沙门菌的垂直传播,从而形成恶性循环。当鸡群暴发沙门氏菌病时死亡率可达80%,对养禽业造成巨大的经济损失。人一旦摄入了含有大量沙门氏菌的动物性食品,可导致食源性中毒,据统计在世界各国的种类细菌性食物中毒中,沙门氏菌引起的食物中毒常列榜首。抗生素一直以来被作为治疗细菌感染的首选而得到广泛应用。但随着抗生素的广泛使用甚至滥用,目前细菌对抗生素的耐药性问题已十分严重,抗生素耐药性正在对全球健康构成威胁。
噬菌体(Bacteriophage或Phage)是自然界中最丰富的生物之一,在地球上大约蕴藏着1031的噬菌体颗粒,也就是平均每1g土壤或水中包含了多达108数量的噬菌体。长期以来,其一直被认为可作为治疗药物和杀菌剂使用。噬菌体对某些细菌菌株具有高度特异性,其取决于噬菌体吸附器官和受体菌表面受体的分子结构和互补性,不会影响生物体或内源性组织的正常运转。另外,即使随着细菌耐药性的发展,噬菌体仍会大量保持。在过去几年中,使用噬菌体来检测和控制食品和环境中致病菌及腐败菌的生长受到越来越多研究者的关注,并且该方法已有许多研究支持。Bmvko LYu等讨论了噬菌体检测致病菌的优缺点,指出利用噬菌体检测食品安全及加工制造过程等方面存在的致病菌具有极大的应用前景。例如在蜜瓜切片、鸡胸肉、白菜、牛奶、鸡皮、豆芽菜等食品中发现使用噬菌体制剂后沙门氏菌菌量显著减少。另外,已从多种食物如碎牛肉、猪肉等肉制品、奶制品、生菜等分离出沙门氏菌噬菌体,因此,噬菌体是食品中的天然微生物。本试验从现代化肉鸡养殖场采集粪便、污水和肝脏等样品,筛选能裂解多株病原菌的宽噬菌谱沙门氏菌烈性噬菌体,并对其生物学特性进行分析,以期为噬菌体防制沙门氏菌的研究提供理论依据。
发明内容
本发明的目的在于提供一株宽裂解谱沙门氏菌噬菌体RDP-NSA-19050,旨在解决养殖场禽类因感染沙门氏菌,发病急而治疗不及时,且因致病菌耐药性而导致用药无效等问题。
为实现上述目的,本发明采用如下技术方案:
一株宽裂解谱沙门氏菌噬菌体RDP-NSA-19050,透过电镜观察,该噬菌体头部为二十面体,长径约60nm,横径约70nm,尾长约100nm,直径约10nm,为有尾病毒目肌尾病毒科;基因全长87591bp,不具有毒力基因和溶原基因,该噬菌体于2020年12月17日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏地址为:北京市朝阳区北辰西路1号院3号,保藏编号CGMCC No.21415。
进一步地,该噬菌体在pH在5-9范围内作用1小时,其效价在1×1010pfu/mL之上,适宜生长在中性环境中;在pH=12以及pH=2时,仍有噬菌体存活,该噬菌体的酸碱耐受范围宽。
进一步地,该噬菌体在35℃-60℃内活性保持稳定,效价在1010.5pfu/mL以上,在90℃高温条件下作用30min,依然具有生物活性,对温度有较好的耐受性。
进一步地,该噬菌体RDP-NSA-19050高效感染变形杆菌,其最佳感染复数为1:10。
本发明还提供了沙门氏菌噬菌体RDP-NSA-19050在制备预防和或治疗沙门氏菌引起的疾病的药物中的用途。
本发明还提供了沙门氏菌噬菌体RDP-NSA-19050在制备预防和或治疗沙门氏菌引起的疾病的饲料中的用途。
本发明还提供了一种用于防治沙门氏菌的杀菌组合物,包括有效量的如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050。
进一步地,还包括用于抗沙门氏菌的抗生素。
本发明还提供了一种预防和或治疗沙门氏菌引起的疾病的添加剂,其有效成分包括沙门氏菌噬菌体RDP-NSA-19050。
一种清洁剂或消毒剂,包括本发明沙门氏菌噬菌体RDP-NSA-19050。
本发明的有益效果体现在:
(1)本发明从青岛即墨田横肉鸡养殖场一发病白羽肉鸡肝脏内分离得沙门氏菌NKS-50,并通过实验确定了NKS-50的致病性和耐药性,以NKS-50为宿主,从环境中分离得到一株宽裂解谱沙门氏菌噬菌体,该噬菌体RDP-NSA-19050对禽类养殖环境中致病性沙门氏菌NKS-50具有强裂解效用,为工业化生产噬菌体用于禽类养殖环境中致病性沙门氏菌NKS-50的防治提供了噬菌体来源;
(2)本发明所提供的沙门氏菌噬菌体RDP-NSA-19050的裂解谱广,能裂解包括斯坦利沙门氏菌、鸡伤寒沙门氏菌、鼠伤寒沙门氏菌、甲型副伤寒沙门氏菌、乙型副伤寒沙门氏菌、伤寒沙门氏菌、肠炎沙门氏菌、肯坦基沙门氏菌在内的致病菌,并具有裂解率高的特点,能裂解具有耐药性的沙门氏菌;在治疗沙门氏菌感染疾病具有很好的优势,效价较高,防治效果好。
(3)本发明所提供的沙门氏菌噬菌体RDP-NSA-19050在防控禽类致病性沙门氏菌中具有良好的应用前景。
附图说明
本发明上述的以及其他的特征、性质和优势将通过下面结合附图和实施例的描述而变的更加明显:
图1为本发明噬菌体RDP-NSA-19050的电镜图;
图2为本发明噬菌体RDP-NSA-19050一步生产曲线的示意图;
图3为本发明噬菌体RDP-NSA-19050酸碱稳定性实验的示意图;
图4为本发明噬菌体RDP-NSA-19050对宿主菌NKS-50的裂解试验示意图;
图5为本发明噬菌体RDP-NSA-19050热稳定性实验的示意图;
图6为本发明噬菌体RDP-NSA-19050遗传稳定性试验的示意图;
图7为本发明噬菌体RDP-NSA-19050入血试验的示意图;
具体实施方式
下面结合附图和实施例对本发明作进一步说明。以下实施例将有助于本领域的技术人员进一步理解本发明,但不以任何形式限制本发明。应当指出的是,对本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进。这些都属于本发明的保护范围。
实施例1病原菌及噬菌体的分离及鉴定
(1)致病性沙门氏菌NKS-50的分离及其鉴定
从青岛即墨田横肉鸡养殖场一发病白羽肉鸡中采样,无菌操作取病禽肝脏,在选择性培养基(SS琼脂)上划线,37℃培养18-24h后,在培养基上形成圆形、扁平、边缘整齐、表面光滑湿润、黑色中心边缘透明的菌落,挑取典型菌落继续划线纯化3次,然后挑取单菌落接种于5mL LB肉汤中,37℃200rpm振荡培养6h,得到均匀浑浊的细菌悬浮液。再通过16sRNA分子鉴定和血清型鉴定,确定为致病性沙门氏菌,将其命名为NKS-50,保存于-80℃冰箱。
(2)噬菌体分离及鉴定
粪便处理:称取5g养殖场采集到的粪便,加入10mL无菌水中浸泡过夜,10000rpm,5min离心浸出液,再用0.22μm滤器过滤上清液,收集滤液备用。
制备混合菌悬液:取1mL菌悬液和2mL滤出液加入27mL LB肉汤中,37℃,200rpm振荡培养过夜,然后10000rpm离心5min,取上清液过0.22μm滤器,收集滤液备用。
噬菌体分离:采用双平板法进行噬菌体的分离,取混合菌悬液滤出液0.1mL和沙门氏菌菌悬液0.1mL混合均匀后,37℃水浴10min,然后铺双平板,放于37℃培养箱培养6-8h后,观察结果,如果有噬菌体,那么平板上会有透明斑。挑取透明斑于1mL生理盐水中37℃水浴30min后,取0.1mL浸出液和0.1mL菌悬液37℃水浴10min,铺双平板,37℃培养箱培养4-6h纯化,照此步骤纯化2-3次,至噬菌斑大小均匀为止。得到一株噬菌体,并将其命名为RDP-NSA-19050。RDP-NSA-19050在双平板形成的噬菌斑直径在4mm-5mm。
(3)噬菌体的保藏
将噬菌体增殖液与60%的甘油按照1:1的比例混合后,于液氮中保存。
实施例2噬菌体的电镜观察
取20μL含噬菌体粗制颗粒的液体滴于铜网上,自然沉淀15min,并用滤纸从侧面吸去多余液体,加一滴2%的磷钨酸(PTA)于铜网上对噬菌体染色10min,然后用滤纸从侧面吸去染色液,待样品干燥后用电子显微镜观察噬菌体形态。
由图1可知,该噬菌体头部为二十面体,长径约60nm,横径约70nm,尾长约100nm,直径约10nm;根据国际病毒分类学组织病毒分类第九次报告,将该噬菌体归类为有尾病毒目肌尾病毒科。
实施例3最佳感染复数的测定
按照感染复数为10:1、1:1、1:10、1:100、1:1000、1:10000的比例将噬菌体增殖液和宿主加入LB肉汤中,并且确保培养体系的总体积相同。于37℃200rpm振荡培养8h后,常温下12000r/min离心5min,取上清液铺双平板测定其效价。结果如表1所示。
表1
由表1数据可知,当感染复数为1:10时,培养8h后,增殖液较为清澈,效价最高,说明RDP-NSA-19050的最佳感染复数为1:10。
实施例4一步生长曲线的测定
取对数生长期的沙门氏菌悬液,按照最佳感染复数的比例接种沙门氏菌NKS-50和噬菌体RDP-NSA-19050,在37℃的条件下,水浴10min,然后常温下12000r/min离心5min,弃上清液,除去未吸附在宿主上的游离噬菌体,如此用LB肉汤培养基洗涤两次。再将沉淀重悬于37℃的LB肉汤培养基中,迅速置于37℃200r/min的摇床中振荡培养并计时。前30min每隔5min取样计数,30-60min每隔20min计数,60-300min每隔30min计数,以时间为横坐标,噬菌体效价对数值为纵坐标,绘制生长曲线,得出噬菌体的潜伏期、裂解期,并计算出平均裂解量。其中,平均裂解量=暴发末期噬菌体滴度/感染初期宿主菌浓度。
由图2可知,噬菌体感染宿主菌后的40min内效价没有明显变化,表明其潜伏期约40min,感染后40-90min内噬菌体的滴度明显上升,之后趋于稳定,表明噬菌体裂解期约为50min。通过计算,噬菌体裂解量约为256PFU/感染细胞,表明噬菌体具有极强的裂解和复制能力。
实施例5噬菌体pH稳定性的测定
用稀盐酸和稀NaOH溶液调节生理盐水的酸碱度,配成pH值为2、3、4、5、6、7、8、9、10、11、12的缓冲液,再用配置好的缓冲溶液将噬菌体稀释成1*1010pfu/mL,将稀释液在37℃下,水浴1h,用生理盐水10倍比稀释后铺双平板。37℃倒置培养4-6h后计数
由图3可知,噬菌体RDP-NSA-19050在pH5.0-9.0范围能维持较好的活性,噬菌体滴度没有明显变化,而且在pH为7.0时,噬菌体效价最高,表明该噬菌体最适生长pH为中性环境。随着pH值的升高或降低,噬菌体的滴度下降明显。表明噬菌体RDP-NSA-19050能耐受弱酸和弱碱条件,具有良好的酸碱耐受性。
实施例6噬菌体对宿主裂解曲线的测定
将培养6h的宿主菌悬液以1:100的比例接种到100mL LB培养基中,37℃200rpm振荡培养2h(OD约0.45,约6.0*108cfu/mL)将菌悬液分装在4个灭菌的锥形瓶中,以最佳感染复数,向其中3个加入噬菌体增殖液0.2mL(约4.0*1011pfu/mL,经0.22μm膜过滤),同时以不加噬菌体的作为对照,取其平均值。
由图4可知,沙门氏菌噬菌体RDP-NSA-19050加入到宿主菌NKS-50菌悬液中,作用1h后出现一定的裂解效果,约2h,OD600值开始下降,持续作用至12h,12h后混合菌液开始变得浑浊,OD值逐渐升高。由于前30分钟,噬菌体处于对宿主菌的吸附、侵染阶段,只有少部分噬菌体开始裂解宿主,因此,其裂解宿主的量远远小于宿主的增长量,因此,OD600在在前1h内,依旧保持一定的上升趋势,伴随着噬菌体数量的指数增加,在1h后,其裂解宿主数量大于宿主的增殖速度,所以OD600值开始逐渐降低,混合菌液达最澄清。持续作用12小时后,混合菌液出现返浊,原敏感菌被噬菌体裂解,被突变的耐受菌所取代的一种菌群交替。
实施例7:噬菌体热稳定性的测定
将噬菌体原液分装到EP管中,分别在35℃,40℃、45℃、50℃、55℃、60℃、65℃、70℃、80℃和90℃的条件下孵育30min和60min,然后用生理盐水10倍比稀释后,铺双平板测定其效价。
由图5可知,噬菌体在35℃~60℃内活性保持稳定,在90℃作用30min,依然具有生物活性,表明该噬菌体对高温耐受性较好,适合在较高的温度下生存。
实施例8:稳定性遗传实验
将RDP-NSA-19050连续传代29次,每代培养基中加入等量的RDP-NSA-19050和NKS-50,每代培养时间6h,然后铺双平板,测定其效价。
由图6可知,将RDP-NSA-19050连续传29代后,其效价稳定在1012pfu/mL以上,说明该噬菌体具有较好的遗传稳定性,适合工业化生产。
实施例9:裂解率实验
无菌条件下,分别取1mL样品和1mL宿主菌菌液(1×106CFU/mL),37℃孵育15分钟,混匀后用生理盐水稀释至10-1-10-3个梯度,每个梯度取100μL涂布于LB琼脂平板上,置于37℃,培养24小时,每个梯度重复两次。同时取1mL生理盐水和1mL宿主菌菌液(1×106CFU/mL)作为空白对照,重复以上步骤。选取30-300个菌落数的平板进行计数。该试验重复3次,取其平均值。噬菌体裂解率=(1-处理组菌落数/对照组菌落数)×100%。
由实验结果可知,RDP-NSA-19050的裂解率达98.8%,对宿主有较好的裂解效果,适合在养殖过程中使用。
实施例10:RTD实验
在平板上划分区域,在每一区域中心处滴加一滴宿主菌(约10μL),晾干,将不同稀释度的一滴噬菌体液滴加到宿主菌的斑点上,晾干,37℃培养16-24h。根据宿主菌生长状况,凡是不能连片生长的,称为CL浓度(CL浓度是可以完全性裂解环境中宿主菌的浓度,可以用于指导生产实践的稀释方案)。由实验可知,RDP-NSA-19050在稀释107倍后对宿主菌仍有较好的裂解效果。
实施例11:噬菌体入血实验
在口服6h后,用PCR的方法进行噬菌体的检测,结果由图7可知,噬菌体在心、肝、肺、肾、胸腺和血清均能检测到,说明该噬菌体可以通过口服进入血液,通过血液循环到达心、肝、肺、肾和胸腺。
实施例12噬菌体动物实验
选择高致病性沙门氏菌NKS-50,稀释100倍后,注射0.1mL,建立动物模型,初始60只鸡,造模成功后,剩余40只鸡,随机分成4组。选择噬菌体作为实验对象;设定阴性对照与阳性对照,分别记录鸡群生长状况,具体分组如表2所示。
表2
实验开始时,肉鸡为15日龄。给药10天之后,对不同组的肉鸡进行解剖。阴性对照组包心包肝现象明显,气囊有许多干酪样物;阳性对照组2(强力霉素)经饲喂后气囊清亮,没有浑浊,也没有包心包肝现象,噬菌体组(RDP-NSA-19050)、阳性对照组1(硫酸链霉素)包心包肝现象不明显,气囊症状明显减轻,气囊基本明亮,基本无浑浊。通过以上数据可以得出:本发明噬菌体RDP-NSA-19050可以用于沙门氏菌NKS-50引起的气囊炎的治疗。
实施例13:裂解谱实验
选择实验室已鉴定好的31株沙门氏菌(5株鼠伤寒沙门氏菌、4株鸡白痢沙门氏菌、4株斯坦利沙门氏菌、1株甲型副伤寒沙门氏菌、1株乙型副伤寒沙门氏菌、1株伤寒沙门氏菌、7株肠炎沙门氏菌、1株汤卜逊沙门氏菌、2株山夫登堡沙门氏菌、4株肯坦基沙门氏菌)和沙门氏菌NKS-50,进行裂解谱实验。具体结果如表3所示。
表3
由表3可知,本发明噬菌体RDP-NSA-19050在5株鼠伤寒沙门氏菌、4株鸡白痢沙门氏菌、4株斯坦利沙门氏菌、1株甲型副伤寒沙门氏菌、1株乙型副伤寒沙门氏菌、1株伤寒沙门氏菌、7株肠炎沙门氏菌、4株肯坦基沙门氏菌和沙门氏菌NKS-50平板上均能形成噬菌斑,说明噬菌体RDP-NSA-19050宿主谱较为广泛,对肉鸡养殖中常见的沙门氏菌均能裂解,具有非常广泛的应用前景。
实施例14噬菌体与抗生素复配使用
噬菌体和头孢曲松钠都采用口服的方式,实验以7日龄发病白羽肉鸡为试验对象,每棚3万只鸡,实验分为4组,具体分组如下表4所示。
表4
分组 | 给药 | 备注 |
一组 | 噬菌体 | |
二组 | 噬菌体+头孢噻肟钠(减半) | |
三组 | 噬菌体+头孢噻肟钠 | |
四组 | 头孢噻肟钠 |
注:噬菌体效价1.6*1012PFU/ml,用法用量:每升对应20000只鸡,集中饮水,2-3h。
用药后,连续7天观察鸡群状况,并统计每天的死亡率,结果如表5所示。
表5
分组 | 鸡群状态 | 7日平均死亡率(%) |
一组 | 正常 | 0.076 |
二组 | 正常 | 0.061 |
三组 | 正常 | 0.053 |
四组 | 正常 | 0.079 |
通过以上表格可以看出,4个实验组鸡群精神状态都正常,死亡率相差不大,说明噬菌体在治疗方面有着和头孢噻肟钠一样的效果。噬菌体和头孢噻肟钠混合使用的两组,死亡率较低,且在头孢噻肟钠用量减半的情况下,效果也优于单独使用头孢噻肟钠的实验组,说明噬菌体和头孢噻肟钠复合使用可以增强治疗效果。
综上,噬菌体和头孢噻肟钠复配使用,在保证治疗效果的前提下,可以有效减少头孢噻肟钠的使用量。噬菌体的合理使用,可以降低抗生素在畜牧养殖中的使用量,减少耐药菌的出现,保证食品健康安全,同时提高养殖效益。
应当指出的是,具体实施方式只是本发明比较由代表性的例子,显然本发明的技术方案不限于上述实施例,还可以有很多变形。本领域的普通技术人员,以本发明所明确公开的或根据文件的书面描述毫无异议的得到的,均应认为是本专利所要保护的范围。
序列表
<110> 瑞科盟(青岛)生物工程有限公司
<120> 一株宽裂解谱沙门氏菌噬菌体RDP-NSA-19050及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 87591
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
atgtcacaat atattcctgt tcctgataca gagtggtcta cacaaactgt aactctggat 60
ggtactgttt ttataattga gttaaagtat aaagagaggc ttgacagatg gttcttgacg 120
ctatctgatg ttgatggtaa tgtattatta tctgaaaaga aatgccttgc agaccagtca 180
atcacaggac gctatgtaat tccttcatta gctggagagc tttttgttga acgaatgtat 240
ggtactgatt tacaacctac cagaaataat ttcggaagag aaagggcatt tgagcttaat 300
tattacactc aagaagatat gagattaatg gagaatttat aatgtctgta aaagatagca 360
ctgctggggc ttcttttagg tgctatcaat tggctgtagg tagtgaaact acagccttta 420
atgataaacc tacaagccat gccaaagatt ctatgcaaat ggactatttc gacaacttac 480
aattcacttg taacgtgtct tatacgtctc agaaaaataa agtaacctcc gatgacacca 540
cttttgaaat ctacaaccta aataaagaga tgagagccaa atttaaaaca gttggtgcaa 600
cagttatgct cagagcaggt tacaccactg gctttaaaag agacgcaaat ggtgacctta 660
ttattgagta tgacaacctc ccattaattt atctaggaac tattgaatat gcttatacat 720
ataagcgtgg tgttgatatg gttacaaagg tcatctgttc caatgataaa atggaaagaa 780
ccacgattaa gacatcaatc tcttataaag caggtacaac acgtaagagt gtaattagag 840
atttagtcaa tagattaggc ttctcactta ttgatgaaga cctttcaagt attgatggtt 900
acacttacaa gaatggtttt agtgtttggg gaagtgttgc agaggcactt acaaaggttt 960
gtgaagaaag tagtctacgt tggtatacat ttaacaagca aatcagagta gtccctttta 1020
acgctaaggc tagacaactt tcttgggaaa tttacccgta taatgttatt gattctttgc 1080
aaggttacta cagaagaact agaaaggttc tgaaaaaaga gaataagaca gttattaaag 1140
ttaaaactgg agttcgttgc aaaatccatt tagacgggag aattaagatg ggtgatagtg 1200
tcacaattag agaaagtgaa gattttgaag gtcagtatcg agtaaaaggt ctttctcata 1260
atcttgactt tactggtggt gcttggacaa ctgaactaga tttagagaag gtggaataat 1320
gaagtcacca gttactagaa tgtctgggta tgtttcagaa tgtcttgatg aatttagaaa 1380
agaactgtat actggcttac cagctattat ccagtctttt gattcaaaga ctcaaacagc 1440
cactgttaag ccactttact ctattaacgg tttacctatg ccagagatta ctggtgttcc 1500
tgttcaattt ccaagtggtg gtggcgcatc tttaacattc cctgtaaaaa ctaatgacag 1560
atgctggcta gctttctcaa tgttaccttt agatgacttc gttgtcaatg acaagaatac 1620
tcagatggaa acaaatatga gaaggacaca cgatatctca gactgtgtag ctttcgtagg 1680
catctgcaca agaacacaga attttaaacc agacccaaca gcagtcagac ttcatttcgg 1740
tgactctgtg ttaagagtta cagatgatgg caatttttat tttgaaggtg atgtacacat 1800
ttctaaaaac ttgtacgtaa cagaagaagt acatggttca gattttatca gtgacacgac 1860
tggtgtgagc tttaatgaac acacgcacca ttattactgg acagaccctg ctggtgaggc 1920
tgatactaca gaggcacaat aatgaaaaca gattttgcat taaatctagg tggtgactat 1980
gttgccactt taggttcaga ttcagtgtat gtggctcatg gtgatttaaa gattactggt 2040
aaccaaatta gaattatccc agaagatgat aaagctactc aggttgctca aagacttcat 2100
atcagatgcc ttttaagggc tggtgaagtc ttctttaaca catctgctgg attcccatat 2160
ttacaacttg ccaaatttaa acagaaaact tctatctttg ataattatat gaaagcttac 2220
cttgttgaaa caagagatgt atccaatatc tataactact cgtcttcaat ggacaatgct 2280
caaagaaaag taactgttaa ttttgatgca actactacaa cagatatttt aacagacatt 2340
acgcaagagg ttaatatcta atggcaggat taactacaac aggattacaa actctaagat 2400
atcaggaaat ttttgataat attaaatcaa gacttcttag agatatttca ccaaaccttg 2460
acgtttctga agatagtcaa ttaggtctct ttctggcttc aattgcaagg tctttagcag 2520
acacccatga agtcctgtca gaaatttatg atggtgggac gattgataaa gcggaaggtt 2580
ttaacctcga tgatattaca gctttaaacg ccgtatacag atatgtggct caggctacaa 2640
gaggccaagt agagtttact ggaacaactg gtgcaacaat tccatctaca actagactaa 2700
gaagcactgc tggtaatatc ttctatccag tttcaaacat tacattaaca ccttcatatt 2760
gtgttgaagc tgttcttgaa gttaactctt tacgaactga tgcagattat gttattatta 2820
ttgataacgt tattttctct tatcagccaa aatcttcaga tacaatcaca gtactattaa 2880
ctgaacttgc cgatgcaatc aatggtggta tcgtagcaaa agccgaagtt atcaatgacg 2940
gttcagcatt acgagtttat aaagatgaag gtgacattat tgccagaacc aaccctatgg 3000
tggtgactgc tacgacgttc cttacattca caaaaattac aactatcaat gatgttgttg 3060
ctgaggaagt tggtgcaatc cctgcattag ctggaacact tatcgaaatt gaaacaactg 3120
tagatggtct tgacagtgta tacaacagat atgacctgac aacaggtaga aatgaagaaa 3180
ctgatacaga acttcgacag agatatttag aatctttggc agttactggt gtaggtactc 3240
ttgatgcaat cgtagctgct gttaaaagag ttcagggtgt atcagatgct tcaggtgtgg 3300
aaaatgatac tgaagagaca agttcagaag ggcttccacc aaagtctttc aagattgttg 3360
tagttggtgg tcaaaatgat aatgttgctc aagcaatctg ggacaccaaa cctgctggta 3420
tcagggctta tggagctatc tttggtacag cttatgactt aggtaactta gctcataatg 3480
tctattttag cagaccaaca cccaagtatg cttttgtcaa agtatcttat tctttatatg 3540
atgaagagag cttaacaatc ccagaagaag atatcagaga tagtattgtt caaggtatta 3600
acgcttacgg tagaactttg aaagttggta atgatgttat ccctaacaga atctatgggt 3660
atatttatga tgtcattaaa ggtgttgaga ttaatgaaat caaagtagct ctttcaaata 3720
accaatcagt acctcctagt gatgggcagt atactacggc aagaattact gttgatggtg 3780
accaatatac tgtatgggaa agtagtcagt acaccattgc taaggagtaa taatgtttca 3840
gaaaattgac gacgtttact ataagactct cgatgaaagg actgtaacac agtttaaaga 3900
taaattcatc tatacaagct tacttaaagc gattactgat gagttgcaga cattggaaga 3960
tgtttcatgg caaatgcaca ctgaaaggaa tatcagaaaa gctgttggac agcaacttga 4020
caacattggt tcattaatta aagtaccacg accacttggt gctgatgatg aaacatacag 4080
agcaatgctg tatatccaaa tttttctaag acgttctgac acaactccaa agtttttaca 4140
gaatgctatc atgactctgt ataacgcaac attttcacag atttttgagc atattacacc 4200
tgtgactgct ggtattgtag tcagggttaa tacaagaaat aacgtcattg atacagcgta 4260
cacattagca aaaattgctg caacaaccat tggttcagca gttattctaa gagacgtaac 4320
tctgaatggt actgcttgga cacctgtaga ggtggctgac tcagccttag caattgttga 4380
tgataaagat aactggttcg ttacagatgc taacaagggt cttgttacta acaacacagg 4440
tggttcttta gataagaact tactaggtag cttagcagat gctggcgtaa gagatgctta 4500
tttcgaggtt gataggacgg ctaatagtgg ctcagtagat tacttaaaag ttaataaaaa 4560
ttataacgct acagataact acatcgttgg taaagaaact gtagcaggtg gtgattatgg 4620
tgttatggct gaagtagctc aaatcatcaa aggtagaaaa gataaatcac agcaagaagg 4680
aagttcttaa tggcattttt aaattggtct acagatgaag tagatgctga tggtaaccag 4740
ctaaaagtat taccaccacc agaaattcag gccactggtt tattaagggg tgagcctatg 4800
ggacgccaat ggtttaacta tatcttaaac tatctcctga agaaagcaaa cggaactgtt 4860
ggtgaagtaa ggtcttttgc cactgagcaa ccagatttag tagctaatgg ctggagtctt 4920
attaagaccg aaacaggtac tgcatcaaca agtacaaaaa acctttacac ttatgaattt 4980
gtaggagctt aataatggca gtaggtgaaa ttcaaattag tgccttgcct caagcagcct 5040
taccaattga ccttagtgat atcttccatc ttaagcaggg tattgaggat aagagatgta 5100
ctcttgagca attacttgct ccacactcaa gcctaagaaa taaccctcac ggtgttacta 5160
aaacacaaat tggtttagat aatgttatca atgctcttca gttagttgct gcaaataact 5220
tatcagatgt tgctaatgtc actgaggcaa gagcaaatct acagattatg tcttcagaag 5280
aggttaatag ccttgttcaa cagcatatta atgataaaag taacccacac aatacaacta 5340
aggcacaggt tggtttgagc aatgtccaga actgggtgac atctaacctt tataatgagg 5400
atgcagataa gtatgctaca gcaagagcag taaataactt gtacaaggct gttcaggctt 5460
cttatccagt aggtactatc catctctcta tgaatcctgc aaacccttct acatatttaa 5520
tttgtggagg tacttgggag ttagtttcaa gaggaagagc acttgtaggt tatgatagtg 5580
attctaggcc agttggtagt aactttggct caagtagtgt tagcttatct agtaacaacc 5640
taccatcaca tagccattca atctacctaa ctggtggtgg acacactcat ggtgctgcta 5700
ttactattga tggctttgat tacggcaata agagcacaaa cagtttcgat tatggaacaa 5760
agaccactaa tagcactggt gctcacaccc actcagtgag tggttcaaca aataccacag 5820
gtaatcacac acatactgta ggtggtcgtt atggtggtga ctctatcggt ggtaaacatc 5880
gtgttcaggt ctcagggact gaacaggttt ccagtgtagc tggtgaccac tcacacgcta 5940
ttagcggttc aacaaatacc acaggtaatc accaacacac tgttgcaatt ggtgctcata 6000
cccatactgt tgcaattggt gctcataccc ataaaggcac agtaactttg cagtcatctg 6060
agcatactca ctcaggtact acaggtacta caggtgctgg tcaagcattt agcattgaac 6120
aaccatcctt tgtggtttat gtatggcaaa gaaccgctta aaaattcttt tacagggggc 6180
tgtacagccc tctttctaga ggacaaaaat aatggcagat tacaaattga gtgaattaaa 6240
ctcaatcgat acaatccgtt cagacgacct tcttcatgtc agggttaaaa agagacctga 6300
aatgctgggt gatgaagacc gtcgaatgac ctatcaagac ttcttagcat cttttaagct 6360
tgaaagattt gttcagattg ctggtagtac tatgactggt gacttaggga ttgttaagtt 6420
actttatggt ggtaaggcag tctttgaccc tacaggctct tctgagatta ctatggggga 6480
tattttaaag acttttaaaa ttaacgcaaa tggtcttaaa ttaactattg cagatgcttc 6540
aaggtcagca actgtttatc atactctgaa taagccaagt cctaatgagc ttggtatgag 6600
aactaatgaa gagaatgacg caagatatgc aagacttgct gttacaaaca cattctctgg 6660
gactcagaac attcaaggtg atgttaactt acttcgcctt agaaaccaaa atgcaaataa 6720
tgcacaatat attgaaggtg tagacctaga tggctcggct agatggttgg ttggtattag 6780
caaaaatggt tctgatgcag tgcagttgta caataacaaa tatgactcag ctttgactat 6840
tgcaagcaat atctctgtga ataagtctct agcaatcact ggtcaagtac aaccttctga 6900
cttctctaac ttagatgcta gatactttac tcagacagca gctaatcaga gatttgcaca 6960
gttagctgtt aataacactt ttacaggtgc taatactttc actaaccttg ttgttaagaa 7020
gaatgctaat gctattactt tgcagaatac tgatgcaggt acagcacttt atattcttgg 7080
taagaagtct gatggaacaa ataaatggta tgttggtaca gattctgatg agacacgtct 7140
gaatatttat aactacatta caggttcaca ggtttcacta ggtactactg ttggtattaa 7200
taaaaccttg caaatcaatg gtcaagttca accatctgat tggactaaca ttgactctag 7260
atatattccg gcagcaacat taagtacaat tgcaagaact aatactctca atacattttc 7320
tgtacgacaa gttatcaact ctgatggtga agctcttgcg ttaaaagcaa aaactgaaac 7380
gacattgttc attagggggt gggctgctga tgggacagct aagtggtatg ttggtaatgg 7440
tgatgccgat aacaataata acttaaggct gtttaactac aaaacaggta aagtgcttac 7500
cattggctca acattcgaaa tgaatgcaac tttagcaatc actgggcagg ttcaaccttc 7560
agattggtct aacattgatg cgagatacta caacaaatca tctgctgata acagatatct 7620
gagagtaaga agcacagatt tcaatactag tggtgctggt aaatgggcta agattgcaac 7680
tgtgaatatg ccacaaggcg catcaacagc agtgattgaa ctgtttggtg gggctggatt 7740
taactttggc ctaattgacc aagcctgtaa gacagaaatt gttcttagga caggggatgg 7800
taacccaaaa ggcttaaatg ttgttgcatg gaaaaactca ccgaataatg atgtaagtag 7860
aattgggtat gttaatacct ctggtgataa ctatgacatt tattgcgcag ttggtaacta 7920
tctagtcgct acaacagcta gggttcagtc aacatctaat gcaacagtac aactgtttga 7980
ggctccagag acatctgaca acgctccatc tggctatgtt gatgggacaa ttgctcacta 8040
ttacactagt ctcttgaagc caacaccttc agatattggt gcatatacta aatcagaaac 8100
tgaccagaag attgcagaag cgattagtga ctctacagac ctgaataaaa tctatccagt 8160
aggtattgtg acatggttta acagtaatgt taaccctaat acagcactcc ctgggttaac 8220
ttggacgtat ctgaacaatg gtgttggtag aactattaga attgcagcag caaacggttc 8280
agatgttgct acaactggtg gttcagattc tgtaacgtta tctgttggta acttaccttc 8340
acacactcat agcttctctg cgactacttc atcatttgac tatggtacga aaaccactaa 8400
cactactggt gctcacaccc actcagtgag tggttctact aacaacactg gtgctcatac 8460
acatacagtt ggtggtcatt atggtggtga ctctatcggt gataaaccgc gtgttcaggt 8520
atttggtgca gtacaggttt ccagtgtagc tggtgaccac tcacacactg tatctggtac 8580
tgctgcctct gacggaaacc atgctcacac agttggtatt ggtgctcaca gtcatacagt 8640
tagtggtaac actggtggta caggttctgg ttcagcattt agtgtaacta accagttcta 8700
taagctgatg gcttgggtaa gaactgctta atcccttatt gactgattgt taagatggtg 8760
ttaatattct ttatgggtat tctcaccatc ttggctgatg aggtgattaa atgcctacaa 8820
tcctagcaat tctcctaaaa aatctaggta gcttcttctg gaaactcatt ttatcccttc 8880
taagtgaata catgattgag aaagtgttct ttaagcttgc aagatacctt gcgggtaaga 8940
cagacacacc tattgatgat gagttcgtag ataacttaga aaaagctttt aagggggaga 9000
aataaatgaa gtggctagag gaagctttta aaaataatat tggtgcaatt gttgttggta 9060
tttttagtgt tattgggatg tataccacga tgcaagtttc aagtggtaag caagaagtgt 9120
ctgtcacaac aaagttacag cagctagata attattccaa aagtaactat tcagctattc 9180
gtgatttaca gtctgacatg agactgattc agctagggat ggagaaccag aaagttcagt 9240
tagagaatgt taagggtgaa aacgcaaaac ttaccaaaac ccttgataaa ttctctgaca 9300
gtgtgaataa tctggctcaa tcagtatcag ccttgcaagc tattactgaa aagaacacaa 9360
agaatactga aaagtaaaat ttaaggctcc tttttacggg agcctttttt gttttttatt 9420
tacctactga caaaggtgct tcaatctttc ctgcatgttg gtagttgtta atgccaccaa 9480
taaaatcact tgcagtaagg tgttttaggt cacttagtgt gttaagctgc acaccaatct 9540
caaaagtagg tgcatgaaac tctttattgt tcatcagttc ataaacctgc ttcatatggt 9600
tctgataaat ctgagtatcc ccaaagacac ctatcaagta tcgtggagtg tatccagtca 9660
tcttgcaaag aacttccaga ataaaaccat aggatgcaat gttaaacgga agtcctaaaa 9720
aggtgtctac agaacgctga taccattgca aatccacttc accctcgtta gtgatgtaaa 9780
tctgaaacag aacatgacaa ggtgctaaag ccattgagtt tgctgcaata tctgctgcat 9840
tccaagcatt aacaagcatg tagcggtctg tgatatcacc cttcatcttc gttactaagg 9900
tctctaactg gtctacaaca caaccattat ggccttcaaa gtttctccac tgaaccccgt 9960
agattctccc acctgcatct tctaaccaat cttgttcaga ggaatagtta gagcttaacc 10020
agcgtttaaa atcgtctgac cagatagtcc agcgttctcc atcattttca ccccaagtac 10080
ggtaacggag ttcaccaagt ttattctcac cattcaggaa ccataaagct tccccaataa 10140
cttgccgtgt aaacacctgt ttagatgtta agagtgggaa accagtacgc atatcaaagc 10200
gaaactgtgg tggagcaaat gcagagataa catctccagt tcgtgtagta cgcagttcac 10260
caacggataa aacatggttc aggatatttt tgtaacttga atcagcttgt gacatatcaa 10320
aagtgtcctt ttacttgtgg aacatagatt tcaaaggttg ctttaccatc atctgatggt 10380
cttgactgta cctttgcaaa tacacggcta tcatacagtt tatcgaagaa gttttcaaaa 10440
ggtaaataca cagtggcctc ttcagtgact ttatggaaaa ctgtatggaa aactacacca 10500
gcatacggta aagcattgac aagaacactt gcaccaccaa ttacaaagac atcttcatca 10560
atagagctgt caagatactt caaaaatgct ccaaaggatt ctttactggc cttagcatac 10620
attacatcat ctttgtcaaa gtctataccc aaatatggaa cagagttcgt taagacgagg 10680
tttgtacgtt ctggtaatgg tttactaccc agagtcttaa aagtctcatt acccataact 10740
accaagttgt ctttagtgag tctcttaaac tcttgcatat cttgcttatg tcgaggccac 10800
ggcataccag ttggagtacc aaactcccca ttttcaccac ttgcaaagat taatttaatc 10860
attttgtaaa ccctttctta atgaaccagt tgatgacatt cttcacagct ttattgctat 10920
ggtatattag gtgtggtgta ggttggtatg aaccataagc ccagaatgat agctgctcac 10980
cacaatcaac acatgtcgta ctctcttcag ctatatgacc attaacaatc acatgaactt 11040
caccttgtaa atctgtacac ccacaatgtg gacatcggat aggtgttcca tcagactcat 11100
agcagtgttc aattttcatc tatttaacct caaaaactgg ctcaagaacc aatataaaat 11160
ctacacagtc atcttcttca tccagaatac catcaaagtt cttaggaacg aagtcgaaga 11220
ggctcataac aaaacagtca gaagtgaccc tgatagtacc tccaatgttc tctacagtca 11280
acactgtacg actttctttt acctccctca gaagattact cacaaagtca tctaagaagt 11340
acttcatcag tgacttaaaa cctttagggt aatctgtacc agagtatgtt tcaatctgag 11400
gcacaagctt aacatttata ctagtctttg ttaagaacgt gttagagaac atactatcag 11460
gtagcttaca aacagtcttt ccagtacctt catgttcaat ctcaacaaaa ccagcttcat 11520
cgccacatgt aactatttta aaagatgctt cattctcata ctgtgaagcc ataaactttt 11580
taagtatctc tttaaggaag tctgcatgag tatcttgtaa gtagatatca atcatagtta 11640
atcacctatt atgggaacag acacttgatg tcatcgtttt taaaaacaaa acttgtgtag 11700
ttatgaccat caacatccag ccaaacagcc tgaacatcta caccatcaag attctggtaa 11760
agctctttga tgaagtggtc ttttacaatc tgactgctag ttgtttccaa cttaatgttc 11820
gagattgctg aacctaccgc catatagccc atgacacgtt tcttctcagt atgcttaatc 11880
ttaccatctt tgtaagccat aatgagattg gaaagaattt ttacaccata atacttttca 11940
aggtgagtag gtacaccgta aaagcctaat gcgaagttct catagtggat aatgttttca 12000
cgtttcatat taatctgcca accttaaatt aaaattaaca ccaaattttt cacacacttc 12060
caacatatac tcaagagaga cattcccagt gaggtggatt atgttggata cacgagcctg 12120
agagatacca caaattttag caacttgctc ttgagaaaaa cctctggatt taatctctct 12180
cttaaaacga tgtgcaacga agtctctcat ttcgtctaca tccattggac aaatataact 12240
ctccatttca gcctctctat cagcctccca atcatcttgt ggggcataag ggtcaaaagc 12300
ttcattcatc tttcttcttc cgtttgcgtg aagtgtcaat caatgaatct atgtttgtag 12360
ccctaacctt ttcacgttca tcgttaagat aatcatcaag agccttttca atctgtgaat 12420
catcttgaca gataactgta aagactgtaa agccgtctgt atcgtcaatt atcttaccat 12480
aaccacttct gggatgtaac tttttaagag ccatatcaaa tacctgtgca gcttctgttt 12540
tctaagaaat attcaacacc atacatagct ttttgcatca aatcttcttt aaactggata 12600
cgatggttga gcatagcctg tacatcaata tcatcatggt gtttctcttt aatgtttgca 12660
gattgcagga aatactctgc aagaacacga tactggtagt cagtcacctt atcggtcatc 12720
tctgtcaaag actcttcaga gattggttta agtaactcac cattagctag agcaacacct 12780
tttacaatct ggcaagtgtc ttctggaaca cctgataagt ccacattggg aagttctgtt 12840
gcattgactg gtaaattcat tgccataacc attgcagcag ctaataaaac ctttttcata 12900
atgacctcaa agttatatta aattttgtat tacattgagt ataaaaataa gctataacac 12960
ctgacttgtc aacagatttt atagcttatt tggcagattt tttataaggc tactgccatc 13020
cttccatacc ttaccttacc acacctaacc atacctaacc aaactacacc atacaatacc 13080
ttgttaaaag acccttttaa agacctttta aaaaggtaag tctcatttta atgtagttga 13140
gactagctac ccatacttta ccttaccata ccagacctca ccccgcctta ccgcaccaga 13200
cagcacaatg cagtacagta ccttattaaa agaccctctt aaagaccttt taaaaaggag 13260
cctccgaaga ggctacctag tcaacttaga aaggttttct gcgttgcttg ttaatgttag 13320
ctttcatagc agctaagtgg ttttgctgtt caatcagtcg tgagcgttct tgatgactaa 13380
gtgccatagt gttcacattc tcgattgctt gggtggcttg ctttaaagca cgacctatct 13440
taccagacat tttgcgtttt gcaatagttg cttgatgctt aggttcaacc acacgataac 13500
caacacccat tgaagcaact aggtaaatct tctcttcttt aagaagcttt tcaataaact 13560
tctcaagtcg gtttagacgt gtaagcgcat aagccttcat ttcatcttcg tacttaacaa 13620
tgtcgccttg ataagttggc ttcacaatac ctaagaaggt gtccatctca gaatgtgata 13680
cagtgtcgcc gtaactaaac tttgctgcta atgcttttgt attcataatt aaacgttacc 13740
ttctgaaata acttctacag aaaaacgacc aaagcgtgga cgccaatcac ctacaccaca 13800
aaggttacca gcattttcca gaaccatcag aagttcctct cgtgtgattt gctcttcatc 13860
gaacatcaaa ccaaactcta cagaccagtc acggaagatt ggacggtagc tcataactcg 13920
tgctgtacca atcttaacag acttagcata gataaagtct ggattcttag caagttcttg 13980
tgggtttgct gggcagttct tgatagtcat tgggaagact acatctgtca gcataatggc 14040
tcggtcaatc accttaccaa gtttgttgag ctttgcacca gacttgatac aagcctcaat 14100
catctcaccg ttcataacaa aacctaactg ctcatcgtag taacaagaag ttaccagttg 14160
actttctgct aagagcgcat agtcttcgtc agtcttttta cgtttactgg acagagactt 14220
gtgatatttt gtcagagggt ttaatgggtc tgacaacgta tcgttatggc tcaagaaagg 14280
acgagtgcca gtgattttga tatttaacag tttcatagca tttagctcca cactttagtt 14340
gattgttaag ttgtttgtaa agaaccctca acagaagact ctttagaaac ttggtagatt 14400
tttaacgtgg ctactaccat ccacccatac ctcacctaac aacacctaac aacacaaaac 14460
tacacccaac accaccgcac cacaccaaac atcacttaaa aaccctctta gaagaccctt 14520
aagtgatgct ggtgagtacc cgttggttta tgctgctcag tatactcacc aaattttcta 14580
tgtcaacaat ttttattcat aatctcgtat aactaaccca acagggaact gtaaagaccc 14640
cttacgagtc atcttctgga actgaaccgt taaaggtttc ccgataaatt cttcaggatg 14700
ctcagcaaga tactgtttct tctcatgagt agttttccat gaaacatcta caaagacgtt 14760
aggaagagtc tctacaacga acttaccatg accacgctta tcagtcttta caccagtcac 14820
tttaaactct tcagtgtgca tcttcttgtg cttaatcaag aagtatgaac ggtgaccaca 14880
ctcatagaaa gattcttcag agattgaacg gtacattgca ccttcaaact tagcttcaac 14940
ccacttatca tgagcttcat caaactcttc ccaagaattt acacggatag tctgtacagg 15000
aacaactcta caggtatcac caagcccaac aatgggtgag ttatcaataa cttcacgacg 15060
ctcaggccaa gattttgaat tgtcacagat atcataccag tagaatttaa gaaggtgacg 15120
gtctggattg tcagcattct taatcataga tacaatatct tctaagtccc aaccatgagc 15180
ataaatctct ccatcaaagt cttcaacttg tggatgtaat ttaagcaaca aaagtaaatc 15240
tggaattagt tctgctggaa cgttgtaaac agtattctca cgagaataag ctgtgaaacc 15300
gactgaatca gcatctcttg aaattctaca acggacacca tcaagttttg gttgagcatc 15360
agcagggaac ttcagatact ttgcatggct aacctttgca gcatcatgag caagctgtac 15420
acctaccttc tcagtattct gtgcagactc ttttgtgtaa gcgtaacctt tacggtctac 15480
ttgtttcttg tacttagcag aaacttcaaa gagagcttgc tgttcagcgt tacgctcatt 15540
ctttttaccg atgttcttag gctctgctgt gtactcttca aacatcatct tgccattttc 15600
tttaccgtag gttgtgataa ctttgtcacc tacagcaacg catgaccaga cgttgaagga 15660
accatcttta ttttgtttgt ataggattgt catttttaac ctctgagtga taccaagtat 15720
tgcaagcatt acatgcgtaa agctctaagt catttttatc tggtttatgg aaacagtttg 15780
acccacaacg acaccggaat agcttaccat caacacgaag aatgaatttt tcaagctcac 15840
catcttctgt aagaactgcg ttcttaaatt tatcttctac agtcattttc tcttaccacc 15900
aatctctacg tcgataactt gaacatcacc agcaacatca aacttatgaa tgatgctttc 15960
cacagtgtaa gtccaaccat caatgcgaac cagctcacct tctcgtggga cgattggatt 16020
acgttgagct atggttgtag tggagtgaac aacatcataa cggtagatat agtttactag 16080
gacttccatc gtgaaaacct cactcaataa ccgtagtatg tgattgacca tttttgccgt 16140
agctataaga tgttgttgtt tgagagtttg gacatctgac aacaacaata tccctactta 16200
tagcgtcacc attgagtttg tagattttgc agtcttgcat ctctggaggg aggctataag 16260
aggcgtcttt gttttcataa gatgggttac agccagccag aaaaattact ccaagtgcaa 16320
ccattgcttt cacacaactc ttaaccttat tgttcacgat agtctacctc tctgtatgtt 16380
tttaaatagt gagtgatgag ttctggtaat tcactacgtt taacgtttga acgaagaagt 16440
gtccagtcaa aacctttacc aacccttccc cagagactgt acattttctt catagcaact 16500
ccagaatatc aatacgttca cctttgagta ctccagtacg gtctcttgca atcttaccat 16560
caacaataag aatgtgcgca gactttaaag agtcaccatt aaaacctttt acgaacttac 16620
taggcatcac tacaatggac atctcctgaa tcaactctgg agtatagcaa ttgtcgtgaa 16680
gttgctgaat aacgtctgca agtgtcattt ttgatgtctc caatactcaa tgtcaataag 16740
ttcttgtgca atgcttttta gattaccttt caagaaaacc ttctcagaac ctgaaagaga 16800
tgaattgtca attaacctgt tcattctctc aaacaacttc tgcttacggt tcttgatagt 16860
ttcctttgaa gcacctttca tcattgctgc atgggttctt ccacggtggc tcataagccc 16920
tcctatacaa gatatagtat aaggtactct tccctgagca ccttgtcaac ttagttttgc 16980
aaactctccg tggagtttaa tagcagcttc acagtaagct ttatgggctt cttcaggtgt 17040
gtcaaacata ccaagatgat agtttttacc ttgtaaacgt atcattgctc gaaacctttt 17100
atttatcttc tgtacacctt ttggcaaccc agattgggac tttcttgacc gattgcaaga 17160
gttttgtgaa gctgtacaaa cccttaaatt caagtaccta ttgtcccaac cgtccccatt 17220
cttatggtca acctgtaagg ttttatcagg caaatatcca tccacataca agataactaa 17280
acgatttgct tggtaactcc ttccatcaat cagcaactga actgacttgt gtccatcagt 17340
atttgttctg tagtttccag ccctcttacc tccaaatcta gagttaaaat ttgcattacc 17400
ttctctaaac ttccaagtaa agacccctat ttcggggtcg tagtccaata gctccttaag 17460
acgaatttct gtaatcattt caacaccatc aatgtacctc ataccagctc ttaccaacct 17520
tacctgtgcc agttacaagc gtctcactct tcaaacctaa attcttagat gcctgaccat 17580
acatccagtc agttatggct ttaacttctt gagtcatact ttctgggcat tcccaactgt 17640
tctcatcgtg gtatgccagc aaaagtctag caccaattgc tggtttccgt ccatttaatt 17700
ttgtcaaccc ttcttcacat gctcttctac aagcaaggtt aattgcctcg ttctgaatct 17760
gactttccga acccatgagc aaataattca ataacttatg tggagattta caccacaacc 17820
aagctccagc aacctttaca aaaccccctt tagagattgc agcagtctta ccaaacacct 17880
cttcaagtgc ctttttagtt gctttgtagt cagcttccaa gtcatctagc aacttcttaa 17940
ttttagggag gcgtacaaag taagtttgct tagttaaagc accttcttcg gttgttgatg 18000
ctttaatcgt cttagcaaac ttctcatcac ctgcaccgaa caatacatct ggtcaataca 18060
ctcgcaaggt gtaccagttc tcttatgaac ttctccgcat ttctacggag tgtcggacta 18120
tatcacgtag tagtaatact accgttgcac ttgggaatgg tcattagctt accaccctac 18180
tctactcagt tcatacatta agtatgcttt tcgatagtct ctgaacgttt ctctgttagg 18240
aatctctata gttatgttat ttgctttagg agtcctaaca gagacttcgc tgctgattac 18300
catatcttta cagacttagg cttccagcaa ttcacaacgt tttacatgcg ccactgacgc 18360
ataaatgcca ttcttagctt tcttacgacc tttgctaatc tcatgaagaa gattctcatc 18420
ctgagtagct cgacaagtca agatgtcctg ctcatcgttc aagctaaagt agatactgtt 18480
cagtgtatgt gcatcagtcc ctgaatagac aacgtacagg tcattctcag agtcatagcg 18540
aaggtactta tcaatctctg ggttgaggta ctcatcaaag tgtttgcagt aatagcgtcc 18600
atcttcttga cgaatgaact caatctcttc tttaccttgt gttaccgctt tggtgaagtc 18660
tttgtcaccc ataaagttac aaagaagaac tagctgggct gagttcatat caactgaaac 18720
gatattagcc ccttcctcgc aaatccacac ctccctcata ggtgctccat agacagcagc 18780
accagacggt acgtttacaa taccatattg tgtcatacgc ccagttgaag taccaaacac 18840
cattgcacca gcactaaggc gaccatcagg acgaatctgg ttcaaccaac ctttttcatc 18900
atcctttgag ttctcaatag ttctgcgtcg gtgcatcaaa gtatagtact tagcaatctt 18960
ctgtccaagc tcaccttcaa tcgtatcata agatgattcg gtaagttttg gtgaagtacg 19020
gattaagcaa ggctcaagca aatctgtata tttcttcaca gaccagttat gctcaatgta 19080
ctgtacacct tcatgttcaa cataactcaa accacatcgt tccaccattt cctgccactt 19140
aggatgtttt gtaatcatct ttttattgtc tttgaaacga caaactttga cagggcgacc 19200
gtctgagtct ttcttgtagt tccagtcatc tggaatccaa ccaactgatt tcaagtagtc 19260
tttaacaact gctacctgag tcatacgaga aacttcgaac tcaattggtg tatatggtgc 19320
atcaatcaaa cctgtgtagc gacttgactc aagttcaaag tggttaacaa cgtgactgtt 19380
atagtacttc acagttttga ctgtttttac tggcttccag tctttgcact tcttaccaat 19440
ctttgcgttc aactcattgc aaattgcacg agcatccttc attgctacaa aaccttcctt 19500
gtactcttca ccagttacag tgttggttgg tgtatagcaa tttctctttt caatgttaaa 19560
aatctttgta gttggcttac caaatggctt gatttcataa gtttgcatct caccgttacg 19620
tacctgctga cgatactttg ttttagggta cttggtaatt ctcttcagtc catctgcatg 19680
accaaatgtt tcaacatact cattccaagc ttttgcaaac tcttctccag tgactttacc 19740
tttggtctta atagttggag gaagatgtgg ttcaacctct gaagcaagct cattagtcaa 19800
cttgtcaagt tccttcacat ggaactccat aagctcttta tcagctttcc aaccattgat 19860
agcttgttga ctcatccaga aagatgtttc tttagctcgc atgtaggttt cataagtgtc 19920
tatgccacac ttcttaagct tgagatactc attatcaagt gcccgtttag ttttggcgtt 19980
aatacggata tcttctacaa cacgagtgaa gatttctgca ttccacacac cccaatgctc 20040
aatctctggt ttacgaacac caacacgagc accccatgca gccaaaccat gagcaccttt 20100
ataacccttt ggagttggtc tatccatcca ctgaacacga gactggataa gagaatcctg 20160
aaagaaatta ctccacggct tacattttgg attatcaaag ttccataaat ccggtgcaat 20220
gtgattgaag acccaccagt cataaccgag gccgttatgg atacaaagtc gttttgcctt 20280
taacgcaaac tcaacacctt cacgtaaacc acccttaatg tacttagtgt acttgtgacc 20340
gaggataggc tcatctgtaa agacccatac aggtggctct tcatcgtcag atttatagtc 20400
tgcaaaggcc attacgtgaa ctttagtgaa ctcaaggagt aaaccatcag tttctgtatc 20460
accaaccaaa tgtaagtttt taaaatctac gttttccatt tttactccct ctaaaatcat 20520
taaccccata cattactgca aaaaattata aaaagcaagt tgacaataca cttgacaagg 20580
tgttaatctt tgcgaagtgg gtttttactt aaaaggttac ttaacagttt aactatacag 20640
atactttaaa actcttaaaa gattatttaa tagcttttaa aaagctttaa agtataacgt 20700
atagaatacg ttaagataaa agattaaaac ttaatagtta cttaaaagag gcttacatgc 20760
agaaaatgtt tgttcaccca gacatagctg actttgttag agctttgcag aagttggaag 20820
aacttgacat tgctgctcaa cgggaatatg ctcaccatca caggagaatg gttgatatac 20880
aacatgaaat agagttgtct gaaaactatg aagatgactt aaaatgctct gtatttgagc 20940
atatgaaaga tgttgctaca gcgagacgta aagctaaaga cacagttgct ttactagatt 21000
ctcttaaaaa gaggttgcaa agtggtacag acctgtgtaa tctagcgtca ctgattaacg 21060
atgttgaaac ttcttgggat aggcactatt atccacgttc tgaaaagaca cttgactttt 21120
cttcatccga aaacttaaaa tgttctagaa agaaacttaa ccaactgaga gagaaataat 21180
atgaatatcg atgtagaaaa gatgcttgaa gaaaaaggta ttgacgaagc gttcttaatc 21240
aatgctgtag aaagcatggc tatgattctt cgtggtaaca actacacaga acagtttatg 21300
cctgcaagct ataacttgcc tgaagacaag tctgataacg gtgcttttgc agaacaggtc 21360
aacatgatgc tttgcgacaa aacacttgct ttagcatcta tcgttgtagg tgttgatgcc 21420
cttgcagagt tcatttttta tgagatgcag aaaaatgact atattgccat gaccactgaa 21480
gaaaaagact tgctactact cgcaggtaat gatgaactaa ttgacctgtt aatttcatca 21540
actagtgctg ttatggctgc tcttgagaag cgaatggaag agaagttgtc acgatatgaa 21600
aaacttgatt ggggtgtaga agaactaaac ccacgaatca aggagattat ccagcaaact 21660
aaagagtctt tggaagaagc tgttgaaaat gactctgtag ctgatataga accacttcag 21720
gtgaaagttg cactgatgat tccagctatc tctacgatga tgaatatcgt tgcactaatc 21780
cagctttcac agatgatggg tgtgagtatt gagttcattg aggagatgac tggtggtgtt 21840
agtatgcaag caatcaatga tgttttggtg aatattggtg cttcaatcat tgaagagaaa 21900
ctacgcatta acttcggtga tgattttgtg aagcaactct cagctatggc tgaaaaagag 21960
tctgctgagc aactacacta agtctttata aatcttacaa gcctccttag ttgggggctt 22020
tttagtaatt aatgggggta aaatgagtaa tattatcgca tttactggaa aggcacgttc 22080
tggaaaagac acctcatgtt ctattgtgaa gaatatctta gaggatgagt acggctacaa 22140
tgttgaagta atggcctttg cagacaacct taaactatct gcatcaaaga tatttgacct 22200
gacttggaat gacctgtatg gtgaaactaa agagactcca caggtttttg acttatcata 22260
tcctgaactg atgtcaaaag ttacagaagc tatggagttt accttcagag atgaaggtta 22320
ccatatggac ttcaagctta tgtcagagtt aaccggacga ttgattatgg agcttaagaa 22380
ggttgctaaa ccgactctat tgacacgtct ggggtttagc aaaaagtata aattctcatc 22440
aagacaaatt caacagattt ggggtactga agttgtccgt aaagtaatgg gcgacaagtt 22500
ctggtctaaa gaccttgaaa aaagaatggt tagattctat gaaagttgct cacttagaaa 22560
ccaatatggt tttgtcttaa tcagtgattt aagatttgac tctgaagctg agtggctgag 22620
tagatttgca caccaaacta ttgaagtaaa aagagacaat gtggataaga tttcatcaca 22680
cgtttcagaa aatgggattt ctacaaaata tgagcgtgac attattcata ataatggtac 22740
tcttgcagac cttgagaaca agctaagagc catcattaaa gtttaaaaga gagaatgaag 22800
atgagagtaa aagacaattt taaagttatt gaccaccgtt tagtggaact ctcatctctt 22860
tccaatgagg taatgattga acgtcttaag agagttgaat cacgaaggaa agagattgca 22920
gatgaaattc atgaactgga caagattgaa aatggattga aggcagaact acaacgaaga 22980
ggtgctaatg tctaaaggtc gtaaattgaa agaggctggt cagtttattg gtcattgtgc 23040
atgtccacgt tgtggttcat cagatgctgg ttcaatctat catcatgacg atgattctta 23100
ttcgatgact tgctttagtt gtaacaaagg tttcccagag tgggattttg ataaaggaca 23160
aatcgtgagc acttattcta ctggttcaga caataaaaac cgtactttcc gtggaatgga 23220
tttagacgat gtaaaagaaa acctagaagc aatggacttg aaagacagga agattcctgc 23280
aaaagttctt gagcgtttag gtatcaaggt tgacattgac agtgatggtg aaattgatgc 23340
gcacttctat ccaacttaca aacgtaatga agatggtaag ctagaacatg ttggctaccg 23400
tgttcgtcac cgttacccag aagaccatcc aaaagaacac ctacgaggta agttaaaaga 23460
cttctcaggt ggtgttggag acattaaagg tgaactggca atgttcggtt catggattgc 23520
tccagaaggt ggtaaccgtc tattcatctg ggaaggtgag atggaatgtg cgacagcaat 23580
ctatatgacc tctcttgcga ttaaggacaa gtctcgtcgt aagaattact gtcacgtatc 23640
tgttccatca ggtgcaaaca ttaagtctat caaagacaac tatcagtaca ttacatcatt 23700
tgatgaaatt tacttgtgct ttgataatga tgaagcaggt gctaaagcca ctaaagaggc 23760
tgctggcatt ctccctattg agaaggttcg tttattccag tatccagagg gtgtaaaaga 23820
ccttaacgaa tggtggacaa agttctataa agagaaagat acagttctgg aaggatttaa 23880
gcagcgtatc tacaatgcac cccgttactg ccctgctggt atcaagaact tcgcagatgg 23940
ttttgaggca atgaagaacc gtggtcagat tccattgatt cctttcccag aatctttcgg 24000
agatttgaac aggctgactt atggcggtta cggtttaggt gagattacaa ctctggcagc 24060
accatcttca gtgggtaaat ctgcatacac tcgtgaaatg atttacactg cttggaagga 24120
taccgattac aatattggtg taattcctgt tgaagatacc tatgaagagt tgatggagat 24180
gctctgtgca atccacctga gcaagcagat ttctgagatt ccttatgacg aacgtgattg 24240
ggatgaattg aagggagcgc acgcagaact atctaaaggt cgtcgtatcc acatcgtaga 24300
ccatcaaggg gcaattgacc aagataactt actggagttt gttgactatc ttgttaatag 24360
cttagactgt aagattatta ttcttgaccc tattacgttg gctctgtcac gttcagatac 24420
ggatgaagag gaagttttgt ctgagctatt gcgtcgttgc aaacgctacc agtatgcaca 24480
agtgaacgtc tgtcacgttc gtaagagtgc aggtggtcag aaggctaact ctgaaggtgg 24540
ggatatctct gaagaggata ttaaaggctc tggtgcgtat ttccagattt ctatgaacaa 24600
cattctgtta atgcgtaaca aggttgaccc agaccctgtt aagaaaaact tgacaaaaat 24660
caagctaact aaatgtcgtc gtcacggtaa gtcaacgggt attgctggtc atacttggta 24720
caatccagat acaggacgcc tcatcaaagc atctggttgc ggagttgaca ttgatggtgc 24780
agcagagaat attcgccaac agtttggtat tggcgaagct gaagaccatt acgatgactc 24840
tttgccacat tatgaggatg aagtgtttga ccgtgagact ggtgaagtct atactgaaga 24900
gcagcgtcaa agctcaacga ttccacctgt attaagtgag gatgcagatg actgcccatt 24960
cgaaactgag tgatagttat aaaagagagg ggttcacacc cttctttgaa gaaaaaagtt 25020
taaataaatt tcaggaagag tgtttgacaa agattgatag gttctataga atgctcctca 25080
tgaaatgcga tggagaggtt gagataaaag aagattttca ccagagcatg attacagtca 25140
tcattgcgct gcctcagcat aatattagtt ggctgtttat aattaaagaa gatgtgtttg 25200
aatatcaagt atatcggaga atatcatgaa acactctaaa gcatttgaaa aagtttttgg 25260
ggattcctta aaagccactg ctggaaaacc agcaaaatac tatgaagaga aacgtgtaag 25320
aactggaaaa actgcacgta aagcagcttc taaagataag cacaacttcc agtaattagt 25380
atttgacaat agagtatcaa caaattagaa ttggtactct tcataaattg agatagaggt 25440
ttaaataatg tctaaagttg ttaaaatgaa agctccggta gagaagtaca acggtactga 25500
acgtcaaact ctgcgttacc tgctgaaaga tgtctggttt tattacctga acacttcacc 25560
acgtccggga aaaggtaaat ctattgataa gaaattccca ggcaaagact gtaactacag 25620
cgtatcaatt ctggcagaag acggtaacaa actgtttaaa gagtttacta agtctaagaa 25680
aaatccagaa ggttgggaca aagttactac tgaagcagtt gatgcagacg acttcgaaga 25740
gaagtttggt tgtaagccac cttttgaagc agacacttac tacatcttga aagtaagtcg 25800
tgcagcagct tataaagatg gtgctgtatg gacagctaaa cagtcattcc ctgtaatgct 25860
gattgaagaa gtaaatggta agcgtgtagc tgttaagcag ccgatgaaga aaatcaaagc 25920
tcaagcatct gataaacatg aagatgacaa gaactatgac gtaattcatc cagatatcgc 25980
agttggtaac ggttcttttg gtagtgtgat tctttctact cacttctaca cttttgagaa 26040
caatgttctg acaaaaccta ttcaggaaca gtttatcatt gatactcttg taccttacac 26100
tggtggtaac ggtgctaatg gtgaacctga acttgatgaa gatgaactgg ctatgcttgg 26160
tcttgatggt gttgaagata acggtgaaat cactgaagaa gacgcaacag accataaacc 26220
ttcgaacgct tctgatgatg gtgatgatga agatttgcca gacccagatg acgaagaaga 26280
tgaagacttc gatacagaag actaatctct aaaagttact ttaaagccct gtacttagta 26340
tggggctttt tcatatggag agccataatg gagaagtaca cattaacaaa acttcccgat 26400
tcagttacac atgtttttat tgactctgat agtattgcct ataaaggtgc ttgtgtagtt 26460
gagaaagcaa aatataaata tgtcaataaa ctcacagcag aagaatctga ttcgtttgat 26520
aatgcaaaag atgctgcaag atggttagca gaccagagaa tccttgtgga agagcttggc 26580
ctgacatttg atgaagatga atgggaaaga cagacttgga aagaagctaa gagtgaaaaa 26640
gaagctatca tggctactca acaggtgctt caggaatggc ttaagattgt tggtaaagaa 26700
agaacttggg taggttattt aacagagaaa ggtgtgcata aacataaaga cattaaaggt 26760
cttgagcacc aatatcaagg gaaccgtaaa gatgctgtca caccaacaca cttagttgct 26820
tgtcgtgagt atcttctgtc aagaccagag ttcaaattga ttcttgatgg gtttgaagct 26880
gactctatcg taattgctaa agctgaaaag atgggtaaaa aggctgcctt aatgagtatt 26940
gataaagacc ttcgccaagc tgaaggcact tactgcattg atatgaccta tgaaaaatca 27000
cctttaattt ttattgctga taacaatgtt ggtgaaatat gggattgccc aataaaatca 27060
acaccaaaag ctaagaagac agttggggta ggttttaaat ttctttgcta tcaagccgtg 27120
gctggtgata atgcggataa ttattttggt ttaaagggtg ttggcaaggt tactataatg 27180
aaagcccttg agggtaaaac cacttacaaa gagtgtcttg atgcaattta tgagctttat 27240
gctaagaaag agtcgtacac ttatgtctcg tgggatggtc aaacaattac cagaacaccc 27300
ttagagttaa tgcagcaaca ctttttctta gcttatcagg aaagaaataa gaaagacgat 27360
ttcacttttg ataagtatgg gtggacacca aatgttaact caacaaactc ttaaagagca 27420
cctgcactat gaccccgaaa caggggtctt tacttggtta gttgatagaa aaggtaaggc 27480
caaagctggc tcagtagcag gttttaaaca ccatagcggt tatatctata taaaactctt 27540
cggaaaaaga tatgcagcac acaggctggc attcttgtat atgaaaggat acctcccaaa 27600
atttgttgac cacaagaatt gtatcagata cgacaactgg ttcgacaatt tgagagagtg 27660
tagcaggtca gacaacaact gtaatgcagt caaaagaaga gataataagt ctggtgtgaa 27720
gggtgtatat taccacaagc aaagaggtag atgggctgca caagttaagc ataataaagt 27780
ttgctaccat ctaggtctat ttgactctgt tgctgaggct gaagaagttg tcaaaaagaa 27840
aagagaagaa cttcatagag agtttacaaa tcacggagaa gctcatgaat gattaaagtt 27900
aaaggttttg ggagttgtcc aacttatgga cattggattt cactatgtgg tgaagttgac 27960
ccagcaaagc actttggttt tgtctaccta gtgtactgca aaaagactgg acagtactat 28020
ttgggtaaaa aacagcttaa cagtgtgacc aaaagaaaag ttgctggtaa gactcggaag 28080
aagatagtca ctaaggaaag tgattggatg acttatgaga cttcttctga gtatatcaaa 28140
aaagatattg aaagctttgg aaaagaattt tttgactttt acattatcca aacctactac 28200
acaaaaggtg gtctagttta tggtgaagca aaccttcaac ataagttcga tgtaatgaca 28260
aaaaggattg actcgaagct cagactcttc tacaatgcca atattgcagc aattaagttt 28320
attaccaaag aaacttatga agatgctgaa aaaagaatcc ataaggtaat gaaagcaaat 28380
tgtgcttgat aactattgag agagaataaa atgtttaaca aaagtaaagc tgtaagtcat 28440
gtagcaaaag ttgacagcaa gattgaagaa cttgagcgca tccttgcaaa tgctagaggg 28500
tcaataatta aagaagttga agctgttgaa tctcaaatgc agcacctaat gcttaaacgt 28560
caagagctac gtgagcatct agaatatatt gaaacacgag agctaaaggt taacaaatat 28620
tttaaggagt ccgaatgttt aactcaagag aatcagtaaa aaactggaac cttcgttgtg 28680
gaaacactca aaagcaacct tacagtgatg agtattggga atctttaaaa tctcaatctc 28740
tgtgtatgct tgaagaagca aaagagcttg taaaagcaat tgaagagaaa gaccccattg 28800
aggcactgga tgctcaggca gatttacaat atgttcttga cggcttgatt tacctgtcac 28860
aacatgacca taacggtgct atgaaagctg tttgtcataa taataacctg aagtacacag 28920
atgattatga agaagcttta aaacgtcttg cagacattga aaagcgtact ggtcaagagt 28980
gtattatcag aatgtcagta gttgatggta aagagtggta tgcgattgtt cgtgcagccg 29040
atgggaagat tatgaagcaa tcaaatctcc ctaaagtaca gcttggtgaa tacattgtag 29100
agcttgaaag ccaagaactt tttgtggtag tatctgagac atgcgttatc tgcaaaggta 29160
ttgtaggtag cttaaaggat ttgggtgtag atggttttgt agaagtcaat ccaattacct 29220
ctaaagcaga taaagatttc tgtaaagaga acggactatg gattgcagat attgtctact 29280
atgatggtga gcagttccat gtaacctcat acccgaaact gaattatgat gctaataacc 29340
tgaagtgctg gttaaaaggg gttggttata atggattcac agaacattaa taaagagggt 29400
gtggctcaga agagccactt ctcaaactac aatatttcta tgacggtgtt tatgaatgac 29460
ccgttactag aaaaatatgg tgagactcca gatacacttc tggacaatga acaagtttta 29520
aaagcagtcc tgtacaaata cgggattgat attgagaaag agtattcctt tgaaatctgc 29580
caacacagga atactttcgg taaagttgtg atggctccac tctttatggg tgtagaaagg 29640
actgactatg gttggttgta tctaaaaaga aacttggaga aataccgtgt ctaaagcaaa 29700
aaagctatct tatgatgaca ttatctcagg tgctaaatta ggtgttgata gtatcggaca 29760
agatgttaag cacggggaca cagttatgta ctgtgatgac cgaagaggca gaagtgcaat 29820
cttgtttgga agaattgttt gcaagatgcg aggtaattac gttgttgcag acatggatgt 29880
gaacgttaca caaaaacttg aaacacttat ggatgataat acaccatcat ggttctccct 29940
gaattgtatg cacacttctt cagtcacaag agtaagtgat aagttttacg atatgtggca 30000
gaatgagcaa attttcaaga tttaaactag ggagcctctt cggaggctct tttcatctgt 30060
aggattcgaa aatgattaag acaattaaaa aatcaaacgg tacagtagta agctttgacc 30120
cagaaagact gaataagtgg gcatcatggg cagataagcg tggaattatc tggtcagaag 30180
tcactatgga agccatgaaa cgtgtctatg agggttgcac tacaaaagag atgcaccaag 30240
ccatgattga tgtttgtgtt gataaacaaa ctcaagagta ctcagatatg gctggacggc 30300
tgcttttggg tattatctac aaagaagcct ttggaggctt tactaaggtt cctacgctgg 30360
ttaccttcgt taaaaatatg gagagagcag gactttggga gaagatggat tattcacagg 30420
aagagcttga atacctgcaa ggttacatcg tacactcaaa agatatctct tacggttatg 30480
cagtcttgaa acagttcaga gacaagtatg gtatccgtga tattaaaacg ggaagacttt 30540
ttgagtcacc acaatttatg tttatgggta tggctatgaa agcctttgag aagcaaccaa 30600
agcaccgtag actgcaagat gttatcaagc tgtacactta cctatctgac ctgaagatta 30660
acgctcctac gccttattta aatggtttaa gagcaactaa atcaggttat gcgtcatgct 30720
gtttgattaa ggcaaatgat actgctgagt ctctcggtat tgctgcaaag gttgcttatg 30780
atatgaccac aaagcaagct ggtattggta tgctgatgga gacacgcact attggtgatg 30840
gtatccgtca aaatacgatt gaacacatgg gtaagctacc ttattacaag cttgtacgtt 30900
catctgtaga ggcaaacaaa cagaagagtc gtggtggttc agctaataac ttctacactg 30960
ctctagaccc gcagattgaa gatttactgc gtttgaagca ccctacaacg gttccttcta 31020
aacgtattaa tgagatggac tactcattcg gcacaaacga ttatttctgg cagtgtgttc 31080
agtatgatac agattggttg ctattctctt acaaagatgc accaaaactc tatgacatgt 31140
tctacacagc atctgctgat gagtttgcta tggcagttgg tcacgcagta cattcaggag 31200
ttaagcacag acgagtaaag gctcgtgaaa ttgctaagct gtttattcaa cagcgttatg 31260
ctacaggtcg tgtgtatcca ttcttcacaa ataatgcgaa cacacataca ccatttaaag 31320
aaccattgaa gatgtcaaat ctttgtatgg aaattgtgtt gccagtgtat ggttttgaga 31380
aagagacaga cctttacaga gacgatgctg tgaaagagga tggtgaggtg gctctttgct 31440
tcctagctag tttggttgca gggagaattt cagaagatga atacgctgac gttgcttatt 31500
atgctcttgc aatggttgac tccgttattg accttatgga ttatccgtat ccgtcgatgc 31560
gtaaccatgt tcagaagcgt cgttctgttg ggattggcct tacaaatgtg gctcattacc 31620
ttgcgaagaa ctacgtgaac tactcttcac gagcaggtaa aacaaaactt catgaactgg 31680
cagagatgca ctcttactat ctgcatgaag cctctctgag acttgctaaa gaacgtggcg 31740
tgccagagta catgaagttc actaagtatc ctgaaggctg ggttcctcca aagacagcta 31800
acaagaagat tgatgaaaag catgatgcaa aactaagata tgattgggat gacttagcac 31860
aacgtatcaa agaaaatggt ggaatccgaa actctgtatt agaagcttac atgcctaatg 31920
agagttcttc actagcaact aatacgacaa atggcttgta cccaattcgt gactttattt 31980
taactaaaaa gtctgcaact ggtaacgtac tgtttattgt tccagattat gaagagttga 32040
agtatgtcta tgaaattgct tgggatattg acacctttga tatgattgat tgttatgcaa 32100
ttgttcaaaa gttcactggt caagctatct cttcagactt ctatgttgac tatgcaaagt 32160
ctaagaaggt atcattggct caagctctga agtacatgat ttatgccaac tcggtaggta 32220
tgaaaaccat gtactacctt aacagtcgca ttggtgtagg taaatctgca ctgcaagatg 32280
cttattgcga gggttgtggt gtttagtttt aacaactatg agggtcgtaa aagaccctct 32340
aaaaataact ttggagagac catgaaagat ttaatagaaa aacatgagcg acctatatac 32400
ttgctccaca agcctcggaa aacgatgtac tatgtgagta gcacagacat gatgataaaa 32460
caaaatgatg actcatgggt tgctggtatc tcttacatct ctacagcaga tggtaaaatc 32520
tatgcaagac cttatgagat gtttaacaaa gaaaattggg aagttttaga cagaaaacaa 32580
gccttagaaa tgataaagaa aggagaaatc acgttatgat taaccagcac ccaatctttt 32640
taggtggtga gagaaaaact tttgactcac ttaataaaca ctacccaaaa atctttgagc 32700
tttataaaca acaaaaagca caagattggt cagaagatga gttccctttt gaacaatcac 32760
gtcttgattt tgagagtgtc ccagcatcaa tgtcaggtgt aatgcttgag attcttaagt 32820
ggcaatggga agcagatact caagttgcta agagcttggc atttgccttt gcacccttta 32880
tctctgatga catttatgca actgcaatta tgaagcagtc tgagattgaa aacctacatg 32940
ctcttactta ctcagagatt gtaagacaat gtattaaaaa ccctgaaaca atcttagatg 33000
agattaacca gaatgttgct gtgcaagacc gattaaaaac tgtgaatcgt gttcttgaag 33060
aattactgga tgaagggata aactaccgac tgagttatgt ccgtgattca cttctggaca 33120
aagacccttt acacttccat aaagtgattc taaaaggtct gtttgcagtg actgcacttg 33180
aaggtatatc ttttatggca tcctttgcat gtacctttgc acttgatgct caagataaat 33240
ttcagggtat tgctcaagct gtccagaaga ttatgcttga cgaaatcctt cacactaaaa 33300
ttgatattga agttttaaaa gaaactttaa gagatgatga gtggcagaaa gcttttcaac 33360
aaattcttcc agagattaaa gtaatcttag atgaagtagt tgaaagtgaa gagaaatggt 33420
cgtattatat cttctccgaa gggcgtgctg tagttggctt aaatacaaag cttcttcatg 33480
agtgggttta ctataatgct gccccactgt atgatatgtt tggcattccc agagattttg 33540
tagctcctaa agaaccacct ttaaagtata tgattaagaa gatggaaatt gataaagagc 33600
agaatgctaa tcaggagcaa cagaacggtg cgtacctatt gaatactgtt gtagatgatt 33660
tgaatagtgg atttttagag gttccttaat gacttatgta atttactcca aaactggatg 33720
ccctcagtgt gagactgcaa agaattttgc aaaagctcgt ggtattgacc atgttgtgag 33780
aatgttaggg caggactatg agctatcaga cctgatggat attgcacaga tgccagttcg 33840
tcagatgcca ttcatcatga aaactgatgg acaaaaccta aaacctgttg ggactctaca 33900
gaattttatg gcagaggtga ataatgctta aacgcatttg ggaaggtttg gttgttgatg 33960
caccagctat tgtgattggt atgcttattg ttaacctatt tactgacttt gaacaaggtt 34020
cattgtttgg agccatgtta ctatgggtta tcttcgaaat attagagata cagttaggta 34080
tcactgaaaa actaagagaa ctctttgcga agttttctaa aaagatttaa aatgaaaggg 34140
tctcttcgga gaccttttta gcatgtaaag gggcaataat gaacaaagta cagattatta 34200
aaaagaatgg ctcactcgaa gaacctgata tcaaaaaagt tttagcagca gtcacaaagt 34260
cagctagtag ggttgggtat aaagaactcc caccagatgt tacccaagct cttgagtcag 34320
catttatgag gcttctggta aaatccacta agcagaataa tttgctaatt tctgtaaatg 34380
atattcacag tattgttgag ggtgctttgg cagaagtcaa tcacgagatt tatgagtctt 34440
actcaacata tagaaattac cgtaaagaag ttgctcaaaa ttgggatgaa ctctaccaga 34500
agaccaaaga tacactcttc ttaggtgaca gagagaatgc taactttgac agcagtttaa 34560
tttctacaaa aggttcaatt attcgtggtt atctgactaa agaaatcttt aaacagtacc 34620
acttaacacc agaggaactt gaagccattg aaaaagggtt tatctatatc catgatttaa 34680
gagacctaat ttttggtggt atcaactgtt gcctgtttga cattggtaaa gtcctgaaag 34740
gtggctttga aatgtctggc attgaatatt gtgaaccgaa gtctgtactg tcagccttac 34800
aggttattgg tgacgtagtt ctttcagcaa ctgcacagca atttggtggc tttactttag 34860
cagagattga taaggtactt gtgccgtatg ctaagaagtc tctacgctat catgctgaga 34920
aagcaacagc ttatggtatt cctaaagaac attaccacaa ctatgtcatg gagcagctac 34980
agattgaatt aactcaaggt tttcagtctc ttgaaatgaa actaaatact gtaccttgta 35040
gccgtggtga ttttgcattc acaactttga cattcgggtt gcttgactca gatatgtcta 35100
atgaagataa ccgactgcaa tacatgattg cgagcactct cctagatgtt cgtatgaatg 35160
gacaaggtaa gagcaagaag cctgttgtat tccctaaatt ggtttatatt tatgaccaga 35220
agagacacaa tgagaatatc tgtcaaggcc acctgtacag taaagctatt gagtgttgct 35280
ctaaagctat gtatccagat ttcttaagtg tatctggaca tggtgctgta gcagaggctt 35340
ttgagcgttc tggcaaggta atttccccga tgggttaaca ggctctatag ctcatctaaa 35400
acgtgcctaa acagggaaac tctacagatg tagacaatcc tgtgctaaat gatgtacgat 35460
gtaagaaact atacaacata ggagtcttac atgccaaact atcacgtcac tgaagatgga 35520
aaagtcttta gggaaaatgg tgtagagctt acgcagtgga aatccaacac tgggtatatg 35580
aaagtgaggt tttacggtaa gaagaatcgt gacatgtatg tccacaggtt agttgctgag 35640
aaatatgtac ctaacccaaa taatcttcca attgtaaaac ataaggacga taacaaacta 35700
aataaccacg catctaactt agaatggggt actcattctg agaatgctaa agaaggttat 35760
gagaacggat gctacaagtt ccacaagcgt tcatatgccg tgaaagccac acacaaagtg 35820
acaaaggaag ttattgttgc taagtcaata cgagagttat ccaatatact tggacacaat 35880
cgcaagacca tctcttcaat attgaagggt gtaaaagagc ataataactt cgaatatgag 35940
tttgaataca tctaaatgcc gaacgactaa ccgtgatgaa tgtagcggtg tagggtcaag 36000
cgactcgaaa tggtacgcta cttagaaata agtagaagat atagtctgga cttactggcg 36060
acagtaagca gcttgaataa agcggggtaa gcttagcgaa cttacctgaa caacaagtgt 36120
agagcatttt tatcaccata ccataacgaa tatggtgaag agttttatgt aggtcgtgct 36180
aacattggtg ctgtatcttt gaacttacca atgatttacc agtattctaa agagaatggc 36240
ttagatttct ggaaagagct tgataagtac ctagaaatga ttcgcagctt ccacaagaaa 36300
cgttacgaaa tgattgctaa tatgccagca agttctaacc ctcttgcatt cacacaaggt 36360
ggtctgtaca aagggaataa gaaacctact gacaaggttg gctgggatat cgtgaagtcc 36420
ttcacagctt cttttggggt tactgcgctt gatgagctag atgttcttgc tgaaggtaaa 36480
cgacttcatg aagttggaag ctgtagtttt gcatatgatg ttctggaata tatcaacatg 36540
aagactgaag agtttaaaaa tgaagatggc ttcttatatg cagtctatgg cactccagca 36600
gaatcacttt gtggaactca gctaaaacag ttcagagata tgtttggcgt tatcaaaggc 36660
gtttctgaca aggaatactt tacaaacagc ttccatatga atgttgcagc cgacattaca 36720
ccatttgaga agcaggactt agaagaaccc ttcttccata tctgtagagg cggcagaatc 36780
cagtatgtaa gagtggctaa tccagaaaac ttaccagcac ttaagagttg tattacaaga 36840
gggatgttga aaggtttcta tcagggactt aactttgact tagcaatctg tgagcattgt 36900
ggcaacagac caaaggctga tgttgaagaa tgtgaggttt gtcattcaca cgatatctct 36960
gtgattaaca gggtgtgcgg gtacttgtca ttcacaaaaa ttaaaggtca gtctcgaatg 37020
aatgacgcta agatggcaga aattaaggat aggatttcta tgtagcatat aaatgaatta 37080
cgaactaata tataatgcaa ttattaagaa ggcacattct gaaaggggtg tgcctcttac 37140
atactacaaa tgtcaaggta agggttatga aagacaccat atcataccag tatgtctagg 37200
tggcgataga attagtggag ataatattgc tttacttaca cctagagagc attatgtagc 37260
tcataaatta ctggttaaga tgtatccaaa caatcataag ttatggtttg catggaacag 37320
gttagctcat gatggcctag ataggaaaat atcatccaga gactatgaaa aggtaagggc 37380
tttgaactcg aaagcttcat ccattgtgaa taaaggtagg gttttcactg aagaacatcg 37440
aagacgctta ggggcagccc aaataggaaa tacaaaatca aaaggttgta aaaggtctat 37500
agaaacaagg aagaagatgt cagaagccgg aaaaaagcca aaggaaaaga aaacctgtcc 37560
tcattgtggc atgactggtg gcgctggcaa catgactcga tatcattttg acaattgtaa 37620
gaaaaaataa ccttatgtaa aacttgacaa agtggtgtag gtcttgatag gcttacatca 37680
cctttttatt ggtgaggtaa ctagatgaat tacatggaga ttagaccatt tgacacagct 37740
aatggtgaag gggttcgtgt aagcctcttc gtagctggtt gtaagcatca ctgtgaaggc 37800
tgctttaaca gggagtcttg gaagtttaat gctggtaaag tgtttactta tgcaaacctc 37860
tacggaatca tccagctaat gggtgataat gctgtcagtg gtttgtcaat acttggtgga 37920
gaacctctgg atgatagaaa tattcaagag gttaccaata tatgtaaacg tattaaaact 37980
gtttacccag agaagtctat ctggctctgg acaggttttc agttacacga aaaaattcac 38040
ctagatgtaa tgaaatatgt tgacgtggtg attgatggta agtatgattc atctaaacca 38100
acagttaaac cgtatcgtgg ttctgataac caaaacctct ggagaaaaga gtatgggtgg 38160
caaggtgatt gtcagtggag agccgagtaa gtctcgtcgt aaagtagttt atcggtaaag 38220
gacactttac aatgttctac agaacaaact taaccgtttg tgtcactgaa aagagcttaa 38280
aacattttgt gaaaagattg agagagatat gaaatgggac tctggaatag ttttaaatca 38340
aaagtaaaag cagcactagc ttcattaggt atgctcactg gtgttgatgt gactcatctt 38400
atgcagaact ttaagatgga tgagaggctg gctaatgaga ttatggaaag tatctatgta 38460
ggccgtggca aaggtggtaa gaaacaagcc caccgtccgt ctggtgcagc agctattaaa 38520
cgagcagcta agaaagctcg taaccgtaaa cgtaataaga aggctaaata atcatgagca 38580
aagtttataa cactcgcaaa ttacagattt tcgtactgtg ccagtttatg gctaaagaat 38640
ataacttcta ctattgtggt acaggtttta ttagtgatga tgatggctca tacctcccat 38700
tcaaagaggc tgtaaaactc tttaatgaag agaaaagctc taagaaagat attgaaaagg 38760
ttaaactaac ctacagcaag aaagataaga agattatctg cctagataac tttgtaaaag 38820
ttactggaga agcaaaagag tttatggatg agagtgagat ttcattcaca agcatcttga 38880
aagtagctca gtaaacagat actataaggg aactgtaaag gttccctttt tattttggag 38940
ataaaatgta tctgtcaaac ctgaaacgtt cggctgcaat gtcagttcta agactcagct 39000
ttgaggaacg tcaagagttc attgactccc accaatacga cccttctaac tctaatcaca 39060
tggttctttg gaatcgtgat aacttccgtg aaagagcact tgtccgttat tacccacact 39120
acacaataga taacctgtat gaatggtgtg ttgtgaagaa cactattgca acactgaaca 39180
atctttgcag gtatacaggt aagcaaacat ttacattagg ccaccacaaa cctgttacaa 39240
agggtggtga gcatcactgt gcaaactggt ttatccagac taagctcgat aaccagaagc 39300
aaggagataa ccttctaagc acacctaaga tgacctatga agagcaagag aaatatatca 39360
aaaataatat gccagatgtg cttgacaaca actatacaga tttggcaata tctctcctgt 39420
tgaagttcga gacagtttat agggcaactt ataatggcta aagagaagtg gaagatttta 39480
ccattagtca gtgaaggtgg taatggttgt gaaatgtaca tgatacgtgg tcatgttcca 39540
gaaccaattg cacttgagat ggtaaacaac tttacagatg gctcttacaa agacttaggg 39600
gagccaacta taaagcagca atgggtaaag cctgtaccag acagcacagg taactgtagt 39660
gtactttatc acgttgtaga ccctgcaaaa tgcaaatctg caatggcagt aacaaacgta 39720
acttttgatt gagagagaaa actatgaaaa catctatccg tgttacagtt cattcaccaa 39780
ctaaagggac tcatgaagaa gagtttaaca tcgtccaatt tccttctggt gagattggtg 39840
gacatttttc accagagttt gttgatttta ctgcttatgc tgcatcatcc atcaacaacg 39900
tgattatgat tgtaaagggt tatgacaaag acacattatt tgctgtggca cttgctaaag 39960
aggcaattga tgatttagta cctcgtaagt ttgctatgaa gactgctatc ttttactatt 40020
tgccaaatgc acgttacgac cgtcacatgt ttaaaggcga tgcagcagct ttgaaagtgt 40080
ttgcacaaca ggttaatgca atgggttttg atgcagtctg tgcagttgac cctcatagtt 40140
acgttccaga taaccttttc aactgcttcc agagtattcc tcaaaaggag attgcagtcc 40200
actatgcaaa tgaccctctg attgattact tagtagcccc agatgcaggt gcttctaaga 40260
agattgcaga gactgctaaa gaggtggata aaccatatat cacaatgtct aaagtacgca 40320
accttaagac tggtgaaata actggtatgc gaatccttga tgatgttgat ttgacagata 40380
aaactgtcat gattcttgat gatatttgtg atggtggtcg aaccttcgta gaagcagcga 40440
aacatcttcg tgaagcaggt gcaaaacgtg tggaacttta tgtaacacat ggtatcttct 40500
ctaaagatgt tgaaaacctt cttgacaatg gtatcgacca tatttacact acaaactctt 40560
taggcgaagc taaagaccgt ggcttaacac attacagtca agttactgta gcaaaccttg 40620
attaggtaac agtatgaata agggttattt tgaaagatac ctaatatatg aaccagatac 40680
aggtttatta atttggagag taaccctttg cagtaccgct attgctggta atgtggctgg 40740
cacaaggtct aagaagggtt atatccaagt ccagataaat aagaagaggt actatgcaca 40800
taatattgca tggattatgt ctggaggtga gataccaagt ggatatgaga tagaccacat 40860
agaccttgat aaatctaata ataagttgga gaatttaaga cttagcacaa agtcacaaaa 40920
tcagaggaat aggggtatat acaagaataa taaaactggt gtgaagggtg taagtttttg 40980
taaacaaaca ggtctttaca aagcaagagt catgctttac cataaagaat acttttgtgg 41040
aagatttaaa acggtagaag aagcaaaaga agctgtaatt aaaaagcgaa tagaacttca 41100
tggtgagttt gcaagacata actaagagag agataaaata tgagtaagtc actttacgca 41160
gtaccagcag gtcttaatgc agatgcttac aaggcatcac atatatatca gtatccagat 41220
gctacgcaat atcttatgtt gaacctgaca ccacgtagtg ataaatggtt taacagtcct 41280
ttagcaattg acggtgtagt ggcttttggt attcaacgtt ttgttaaaga ttacttgata 41340
gaccactgga acgccacttt ctttgaacgt gataaaaaag aagcaattga tgaaatctta 41400
gaagtcatga atggtgttct gggtaaagat gctatcggtc gagagcattg ggaagcactt 41460
cacgacttag gttatctacc agttgaagta tacgctgtag aagaaggcac agttgtccct 41520
atgcgtgttc caatgattgt cttccagaac actgtttcag gtttccattg ggtagcaggt 41580
tatctggaag atgctttctc tgctgagatt tggaaggctt gcaccattgc gactatcgca 41640
ttgcactaca aacgtatctg taagaagtgg gctgacctta cttgcgacaa cgacttacat 41700
ttaccttatc agtgccatga ctttgctatg cgtggtatgt ccggctttac tgatgacgca 41760
tttaatgctg taggtcactt aaccagcttt aaagggactg atagcttccc tgctatatac 41820
acagctaaac gcatctatgg acagtcttac ccaatctctg atattggtag ttctgtacca 41880
gccaccgaac actctgtgat gtgtgcaaac attgcttggg aaggtggtaa cgagttggtt 41940
gaagaagaaa gacgatttaa aggtgagttg caaaccttcc gtcgtttctt aacagaaact 42000
tacccaactg gtattgcaag tattgtttca gatacttata acttctggag aactgtgtca 42060
gaaatcttac cagaacttcg taaagaaatt atggaacgtg atggcaagct ggtaattcgc 42120
cctgactctg gagaccctgt acatattgtc acaggttata aagcaatcca cttagagtgt 42180
gctaaaaagg cttactacga gcacctaagc aagctagagg caagtgatac aatgctgaac 42240
tctgtcctga acatgaagct tgaaaacatt agttacggta ttgctggatg gctactgtca 42300
gaaggctacg aaatggttgt tggcagagaa gactttgaag ttgctgatac ggtgatgttg 42360
aaaaatgctt atatggttgg ttctgcaaac gttgtaacac gtcctgtagc tgagattgat 42420
ggagctatta agacactgta caacatcttt gggggaacta tcaactctaa agggtttaag 42480
gtactggatg agcatattgg tcttatctat ggtgactcta tcacgctgga acgtgcaaac 42540
gaaatcctga agcgtctgta tgaaatgggt tttgcaagct ctaacgtagt gtttggtgta 42600
ggctcttaca cttaccagta catgactcgt gacacctttg catttgctgt caaagcaact 42660
cttgcaagca ttggtggtaa agagattatg cttgcaaaag accctaaaac agatagtggt 42720
gttaagaagt ctgcctttgg tggtgtagca cccatgtggg atggtgataa gctgaaagct 42780
gtagatggct atggattccg gagttttgca gatgtacttg accacccagc ttgtgcttta 42840
cgtttagtct ttagcgactc tgagcagttc ggttatacaa ctcttggtga tattcgaaac 42900
aatattgaca agcagcttta aaagtatatg ataagaggct cctacgggag cctttttaat 42960
ttctggagaa agacatgaaa atcaaagaga tgaatattaa cgttgtcttg gaagaacgtt 43020
gggaaaacat taagaaacct gaaaatggtc ataaattcct taacaaaatt ttagtagcag 43080
ctaaagaaga actgacgggt aaggttgcag cagcaatcac aataaaagtc tgtgtaaaag 43140
gtcttccaga caaccaccaa tttgcacttg acgagtttaa agaaagcttc tacaatccaa 43200
acaaacagat acttgaaagt aactttgcag tgtctacaag tatcgtccat gacagaagct 43260
ttatccttta caaaaacatg agaggtgagt catgcaagca tattggatag aaattttact 43320
gtcacttggt agtgtagcag tgtttgtcta ccttctttgc aagtactatg cagagcacaa 43380
gaagtgtgac tactgtaacg gagaagggta cacaagagca ggttgttgcc ctatttgtgg 43440
tggttctggt aaaatgttta ataagtaatt taacagcaaa ctaagaggaa agtattatgc 43500
gcatggtaaa cgaccacgca gaagtgatta agagttcaac ttctttggag acatctcaag 43560
cacagattac aatgacacct gaaatgttca gccttctgag ttctggtgta tacacattca 43620
aagaaagggc agtgattcgt gaactgtcat gcaatgcagt agatgctcag aaagaagctg 43680
gaaaagagaa catcccattc catgtgcatt tacctactcg atttgagcct tactttgaag 43740
tccgtgactt tggaactggt ttaacccatg acaaagtaat gagtttgtac ctgaactacg 43800
gggcttctac gaagaacgac tctaatgact acattggtgc aatgggtatc ggctcaaagt 43860
caccgtttgc aattgctcag tcattcacag tgtctagcta tgttgacggt gttgttaata 43920
aatactctgt ttaccttgag aatggtatcc ctcaagtgac taagcttact actaacccaa 43980
caaatgaacc taatggctta gctgtacgtg tggctgttgc tgaccatcgt atttcaaagt 44040
tctttgaaga ggctggtaac gtatactcat actttgctgt aaaaccagaa agtaatattg 44100
tatatgatga tgtattagca gatatgaatg tcattgcccg tgaaggaggt gtttatgatg 44160
ctatgattca taagcaaagc tggcgttcta gtggcaacag gacagatttt aatgtggtaa 44220
tgggtaatat tgcctatcca gttaatatgg aagcattact tggtgatgat ttcttcaaag 44280
tcttgccaga atttttccgt aggagcgtag accttgtaaa catttacatg cctattggtt 44340
cagttgctat tgcagcttct cgtgaagcat tgcagatgaa tgatacgaca aaaaatgtta 44400
tcattgaggc tactaaaaag ataactgaag caattacaaa ggatgttatc aagaaagtta 44460
atagccaacc tacacttatg gatgctgcac aggcttatgc ggagttgcgt ttgaactcac 44520
gagaaatgtt taatgctgtg tgtccaaagc tagagtgggg tggtgttaaa cttgatactc 44580
tggaagaaga attgctaaac attcgtcgtg ggattatcta cgcagaagac ggctcagtca 44640
tttatgaacg tgacggtaaa ggtaatatta aggttgatag gaacggaaac aacatcccta 44700
aagtagatta tctttataac ccagttgctt atgtcaagtt caactcttta gagagtaaga 44760
ttcgtgcaac agcactttcc tacactcaag aggcaagtat gttcaatatc tttggtgcaa 44820
tgcgtaaaag ccaaattgaa cagtttttgt tcgtgattaa tgaccgtcgc aataaaaacg 44880
gtactgagaa gactgtagga cgcaaccaga ttctgcgtgg tgcatgtcga gactatgcta 44940
gtgagtcgag tatgtttcac agatacaatg gtattgtctt tgtattctcg actgaaaaag 45000
agttagacga cctaatcaat ctacacaaac ttgataaaag cttgttaaag attgtgaaga 45060
tgtctgataa ggaacaccat tatcaacgta aagaagctgt aagaggtgtt gtaaaactct 45120
ggaaagctgt tccagcagaa ggtgtagctt catacacaga ggtgtcagaa gacttagaca 45180
caattgaaga gcctcaactt tatatcaagg cagtaggtga tactgttgat ggtgaatgtt 45240
tttgctcatc tccagatgat gtagctaagt cagttgccaa tgtcattgga aagacagtct 45300
atgttttccg aaaagcaaat tggaaaaaga taccagaaga ctggattgaa gtagatgaga 45360
agctcttgaa cgacagctta actgatgttc actggattaa tcataacagg tatatgacac 45420
gtatctatat gaatggtgtc ctagacctta caagcagttg gattattgcc agaaacttta 45480
cgttcaataa caggaagatt tctcgtggct attgctattc acgagacaca aacaaaacta 45540
tcttcttaga aggtaatgaa gatgctgttg aggcaatgtt tggtaaaatc caatacgttg 45600
ctgcaccgtt tgcatacact tacactatta gcgtgttgca agatttgaaa aggtgtcttg 45660
acaatgatac aaagctgtac aagaaaatta agaaagctgg tgaccgtatg gtcattaagg 45720
tgacaaacta cctttcaaaa agaaaacaag aaaacttctt gctttctcat ttagattgga 45780
ataaagtgtc acccatcgaa gtgagtaagt tcttaggctt tgatgtgaag tgtgttcctg 45840
aaggtacaac aatttacgat taaagtgttt gacaaggggc ttcaaagccc cttagaataa 45900
cctcacaaaa tgatttatta actaacaaga gagagtaata agatgactac taaaactaaa 45960
gcacagattg acgcagaaat ttacgaactg gttaaagaag gtaagctgac taaaacagct 46020
atcgcacaga agttcaatac ttcaacccgt tcagttggtc gtgctgtaga gcgtcatgaa 46080
gcaaccttaa aaggtaagaa aaaggtagct tctaaaccag ctacgaagac tctgaagcaa 46140
gttgctaagt cagttaaaaa gaaagctgac aagccagtta aaaaggctgt taaagaggat 46200
gtacagaaaa ccccggtaaa taaactgcat gaagctatgc agaatggcaa ccagattgag 46260
tacatgatta ctggcgactc tgttatcatg actttcggtg cagattctga aatcgttgaa 46320
tctacccatc cgaactacaa agaaatcgtg atggctgttg tatccggtga gtataagaaa 46380
gctttcgaac tgatgaacat tcgtaagtct atcgaaaatt tcactcaggg tgctatcacc 46440
atcaaaggtg ataagctatt ctacggtgct gttgagatgc gttctactct ggttgaccgt 46500
attcttcaca tgatgaagac tggtgataaa ggttttgaac gacttgtaat gttcttcgaa 46560
aaactgatgg aaaacccatc taaagactct gtagaacaac tttgggggtt tgtatctcac 46620
cttgatgttg aaattgatga agaaggctac atcattggtt ggaagaaagt ctctactcgt 46680
gaaggcaagc tggttgactc tcacacctac aaagtaccta atgatttggg taacattgta 46740
gagatgccac gttggatggt tgataacaac cgtaacgtaa ctcgctctca aggtcttcat 46800
gttggtgctt gggattatgt tcgttgcttc tcaggtgata ccatcctgaa ggttcgtgtc 46860
catccacgag atgttgtatc tgttccaact gattacaacg atatgaagat gcgttcctct 46920
cgttatgaag ttgcagcaat cgttgataat caacgtaaag tgctgaaagc atgggatggt 46980
aagactgaag ctttgcatgt catcgttggc actgctggtg aactcatctc ccaacgtaaa 47040
cgtgaaatct aataagtaat ttcttaaaag gctgcttcgg cggccttttt tatttgtatt 47100
ttgtgcagag tgctgtataa ttgttgtcac gataaactaa acaggagaac caatgaagaa 47160
gttgattcta ggtttgtgct taatgtttac agcacactta tcttatgcag ttgactgccc 47220
agagctatca attagtcaaa aagtaaacat gttaaaagct taccaatatg gtgagaacaa 47280
catgggtaaa ggttggggta tcactctagc agctatagcc ttacaagagt cagagttagg 47340
tctgaatatg gagaataaaa agacccatga ctatggtatc ttccagaatc acttgaagac 47400
tgttgtaaag cgtaacaaaa tcagtcctaa tgtggctaaa aagaaactct taaaagattt 47460
tgactatgct gcaaaggaaa cacacaaaga gcttgagttt tggacgaagg tacatggtca 47520
accaaagtca aagaagactt tacaaaaagt tttagcatca tataatgctg ggtattcgta 47580
caaaattcct aaagctaaga aatactctca agatgtctat aacaacatga aagtcatcgc 47640
tcaatgtgaa tttgcaacaa acatttctaa ggtaaatcat gaaaaaatta agaaagtctg 47700
atgaagtact gtgtcatgct tacgacttgc atcctcatga gttaggtctt gattcctgta 47760
tatggactcc agaacagtgc agggattttg aagacactgc aagagaagtt gtatgttcac 47820
ttgaagagtt ccacacatca gaaccaattg tgaacgttgt ggacaaagag actgggcaat 47880
ctataggtgt aagacgtgac agtttagtca tagtcaataa agaccttgta gaaaaaggta 47940
acctcatctt agcagatatt gatggtgttc ttacaaattt taatcacgaa gattgctcaa 48000
cggaattgac tgatgggtca ttctcacagt acactaatct tcttgactct gtaagagcaa 48060
aaccaacgta tgtttttagt attattgatg caattgctag tcacgctgcc attggactct 48120
tgacagcgag aggagaatct cagagaatac ctactgagat gtttttaagg cataacatag 48180
agcacgatta catgcttttt atgcgtggtt ttggaactaa ctctataaat gcagaaagtt 48240
tgaaagttag gatgattcag tcttgcattc ttccttactt taatatagta tgttttatag 48300
aagatacaga gaagaatgtt cagaaggtaa acagaatcct tccgcacatt aaaactatgt 48360
tagtaaaaca ttgagagaaa acttatgaac aatatcatta cggtagcact ggacgttacg 48420
gcaaacaaat ctgaagtagt tcgtaacatt atcaaaagca actttgaggg taaaatcttc 48480
cgtgctgtca atgtaaaagc agatggtaac atccgagaat accgtgctct tctgaatgtt 48540
aagaaacacg tcaaaggtgc tggttcaacg actgcacaca aagaaaacct gatgactatc 48600
tatgatatgg gtatggcttc agagttaggt gctgaaggta tcgttaaaga aggtgctccg 48660
tatcgctctt tcaatctgga aactgctctt atgctttcct ttacaagtgg ttctaaaaca 48720
actacttatc tctttactga tgctgcaacg gtatctgcta tcaaggataa cgctatcaaa 48780
gctggggtag ctgctgctgc aaaagcttct tcgatggctg caaatgtcct tgctaaagtc 48840
ctcggttaag gttaagatac aggctccttc gggagccttt ttcatttcag gagattgtta 48900
aatgacattt aaagagttct gtcaagctac tttcataatt gttttcttag ttggagcagg 48960
tgtctgggga ggatattctt acagagacta tcaagttgct gaagctgagc taaacaatga 49020
gaaactaata agcgttgcta aagatgctta tcaggaaggt ttagccacac taagcgccaa 49080
ttacaaaaac gatttgaaag atgtgcttgc taagaataag catacaaaag aggtactaac 49140
atatgaaaaa actaagccag agttttataa tgtttgtgtt actgataatt atgtcagggt 49200
gttcaacgaa caaagtgaac agtacattca aaaacttcca agtaagtgag agtgacaagt 49260
acactcaaga agaatcacgg tatatcatca aaggtaacac aggcagtgat gtggcagcag 49320
cccttgagtt ttatcgtgat ggtttttacc agtgtacaat aaaagctaat aaccttattg 49380
atatgatttt gttaggaaat aaacagcaat gacagagaat gaagatactt tctatgtaga 49440
agggtactta ctgctaccac gaccaaaaga aacttatatg cgggttgatt tctcaccaac 49500
cattatggat aacgtgatgt gtcatatctt tatgcaaggt gtcacagcac aacttaaaca 49560
tgttggtaaa gattgcaaaa taaaggttga cactcatcca gaaatcaaag agaatcacta 49620
cacatggttc ttaccagact ctaaagaaat cttagcagtt cttaaaacga ggcagtaact 49680
atgcagatta atggaagaga ctttgtagcc atttactacg agaaagataa agaagtcggt 49740
gtagcacaag tatcttatgg taatggtaag tggttgtatg ggacaatcgc agtagtcgga 49800
acaagaagtg atacaaaaac ttttaaagat tgtgttgagc ttcttgaaga atccatccag 49860
aatcattggt gtctgatatg gatgactgac aatgaagtga tggaacgttt caaaaagatt 49920
gatatcaaca tagacagcat cgagcatgtt gacttgtatg aactgactga gaaggtaaat 49980
tatgaaagta attctagcca gagataaaaa gacacggaaa attatccggt cagcagtatt 50040
tattggaaga aatgaagtga ttccttttac agcaaatgat gtgatagtct acaggaaaaa 50100
attagcattc agttattgct tacgtggctt tatctcccag ccattggaag actttaaaag 50160
tcgttgtggt aaaggtatca tcattcaaga ggtaattatc aatggctgac ttctgcaaag 50220
actgttctat cgaaatgttt ggacgtgata caggtgactt aaaaggtctt atcactgaag 50280
acgactttaa agctggttat gcaatgccag taatctgtga aggttgtgga tgtatctggg 50340
tagaccacga aggacaacgt gtaaaacctt cagaagataa agaatcttgg gagagatgct 50400
aaatgggtat tgtaaaaatc atcaatgggg atatcttctc cgcatttgat aaaggtaagt 50460
ttgatatcat tggtcacggt tgcaactgta tgaacttaat gggtgcgggt atcgctaaca 50520
agatttctaa actttaccca aaagcatatg agaccgatac agaagtttat ctgtatgcag 50580
gtggtatagg ccacaaaccc tgtgaaaatt tactaggcaa cttctctgta gcacgtttaa 50640
aacaaggccg tatagctaac ctttacactc agcttcacac tggtaaagat gcccgataca 50700
gtgctttaga gtcatctttg aagcaactta acagatactg tgaagttaat cagttgaaga 50760
aagttggctt acctatgatt ggtgcaggta tcggtggact tgaccctcaa gctgtcacag 50820
ttatcatcaa tcaggtgatg aagagtgtag atgtgtatct gtatgtctat gaaggagaga 50880
tgtaccacaa gttacactca ggttggaaga actactgtga accagaatac tttgcgggtg 50940
tagtagtgtt cacagatgat acagttaccc tcttcagacg aagaaaaggt aagatacatc 51000
aaagtaaccc tccggttgaa aagatgtctc taagtaatgc tctagttacc cacctgtcga 51060
agagcaatca caggattgca gtaacatttg gcagtgatgc agatacatat atctatgcaa 51120
gaactgatga ggatgttaag gaaattttct cttcacagga agttaccttc ttagacgcaa 51180
agagctaaga aactttgtag aatattcaca atagactaag cctttcttat ctggtaaatt 51240
tttcaggtaa ggagggcttt ttcattttag gattgtgcag gtagagttct ggacaaaaat 51300
ctgtagagag aattttgaac tccaaagaga aaattcattt tcaaactgtg caggtaatct 51360
ccaaagagct tttttatttt caaaacgtgc agaccccgct caggtcgggc aagaaaaatc 51420
attttagaat tgtgcaggta ggtgggtgta ggtggggatg tcgccatttc tcacaaggct 51480
accacatctg aaaataaaag tcaacagaaa aatttgtagc ggattgaaaa ataattcttg 51540
attgtagttc tgttttgtgg tagtcgcgcg cacccgtttc attacaattc ttgcaaaaat 51600
atttttaaaa ttcctattga ctctttgagt atggttatct aatatttgca tcagacgggc 51660
aagcaagccc acttcctaaa ctggagaatc caaaatgatg tacagtcgcg aattgatggt 51720
tacaaccaac gttaacacaa aggcaaaacg tttttatatg caagtgtgcg gcgtatggcg 51780
tagaatatca gaagctgatt attgcaacta tgtagaaacg gcaattaact ctaattgcta 51840
cctaacaaag aatacaaagc gacatataaa acagacagcg gtttactatt acgatatata 51900
aaaagtactt tacaaggggc ttaaaagctg atatcttaag ccccagataa agagctttac 51960
taacttacct taaactagga gattctaaaa tggcttatcg tgcaccgaaa ttcatcaata 52020
aagacaactt ccgcaacgca ctggagaagt cacttgatga aaaatttaac ggcaaattga 52080
ttgttgttca cgcattcaat tttaaatatg atgtcaatgg taatagaatc aaccattaca 52140
cggcaaccat gttagacggt actctgtcaa gtgaaaaagc cgtactgaat gcacttgctg 52200
gacgcggtaa aaacctgata aaatgcgata agcgacgtta tcagggtggc gcatatggtt 52260
acgatgatgc tgtttaccat cttgaaaata tgggctatca agtagaaaaa gctggggtat 52320
cacaaatcat cgggagtgat ggttatgtga caattttcaa aattaactaa taaagtactt 52380
tacaaggggc ttatagttcg ataaagtaag ccccagataa agagctttac cactaaatcc 52440
taaattggag aacttaaaaa tggctattat caaaaacgtt gtaattactg caaagactcg 52500
tgatgacgct cgcgttatgg ctaaaaagtt atgcggaaaa gtggttgaca acggaaaaca 52560
aactgctgta aggtggggcg tgaaagctga taagcaacta accttaaaac gcagttcaaa 52620
caaccttttt aaaggtgtaa aatcactcgg taaagtgaac gtttttacca aaaaaggcta 52680
taaaatgcac ttatcattaa ctggcaattt aatctgataa agtactttac aaggggctta 52740
aaagctgata tcttaagccc cagataaaga gctttaccac taaatcaagt ccttaaacta 52800
ggagattcaa aaatgaaaac tgtagacgct acctttgaag ttgtaaaaga aaaattttcc 52860
atcatttatg aggctgttaa cacttatcaa ccatcttatg taatggaaga tagctgggaa 52920
tttactgata aacatcatgg cggggtaact gttaagaatc ctaactatga acgcaatgga 52980
tacaaatatg ctattccttt aaactatagt ttaagttgcc ttgctagtga ctatgcaaaa 53040
caaggccgtg aaaatccatc attagaagct tatgcatcaa tgcaaaaatc tttaaagagg 53100
gatttagagg cttcagaata ctaccttgct gcaaaagttg tagatgcaaa cggtaacacg 53160
gttttagata gttttagtat tggatatgtg tttgattggt cttacttgga cggcgatgat 53220
ttagatgaca ggctaaaaga ggaagtgaac aacgcagacg cagaatctga agtattagaa 53280
cgcttagaag ccttaaaaga ttcagtgatg aatatcttta aaaattaaat taagagggaa 53340
catcatgtta atgatgggta tcactgcaat catcgccatt atagcacttt acaaggctta 53400
cagcgcctac aatctggcaa gtgaggcaat tactcaacag gttaataata accttgtagt 53460
gagctttctg gagcgtttaa gtgatgaaca acttaaacgg ttagaaatga gcttcagatt 53520
tcaatcaaaa acttatcaaa tcagcgacat tttcaaagag gattttcagc ttgtcgatga 53580
ttataacgcc ctgataaatg ctttaaatat cagtgactta aaagactatt acgctgtaat 53640
tgttcaggag ttgagcaaac gaaaacaaaa cagttgacac tgatttttag atatattaaa 53700
ttacacatca acggggaggg aatgaccttc cccactaagt aaagtcctta aactaggaga 53760
ttcaaaaatg tatactacta acaatggccg tactctaaat gtcactctac gccactatgt 53820
agacggcgtg atgaactttg aggatttaca agcggaacaa catattttgg attggcaact 53880
tgctggactt caaaaaactg ctaccggata tgggaaaaag ataccaacca gctggaaagt 53940
atactacaaa ggccgtttgc gcagaattta ccaagatgtg tgcagcaata gcgcatcaag 54000
ttacatcatg gtaaaaggca aaaagctgca tttagtgtaa agtactttac aaggggctta 54060
tagttcgata aagtaagccc ccagataaag agctttaccg ctaaatccta aattggagaa 54120
cttaaaaatg gctattaata accgtgaatt cttaatctta aaagcacgtt tgaccgtcaa 54180
ccggattaat gttatcacgt catcagcgcc agacgaaaca ctgcaaaaca ttatcgggaa 54240
gattcaaagt gttatcttag acgttgaaag cgtaaaaaac tcactgattg acgttgcagc 54300
gggtatcacg ctagacggcg cacaatatga aatggctgac atgttaggca aatctaaggt 54360
aatgaataga gagctggatt tgaaaatgtt tcgttttgct gttaaagtat ggctatccgt 54420
cgagtatggc gcaaattttg caatcgctga ctttttcgca acatggttac aacgtaattt 54480
aacaaatcac gattttcgtg atatctgcga cgtaatttat tcagaacttt aaaaaattcc 54540
tcttgatttt ggtttttaaa acaagtaaat tactaatcaa cggggaggga atgaccttcc 54600
ccactaagta aagtccttaa actaggagat tctagaatgg ctatcattaa cggcttaaac 54660
attgaaacta ctcacatcaa agatatcaaa gttggcgata tagtcctttt tcacggcgta 54720
gaaaaaactg ttacttcaaa ggatatcaaa gaggactctt tcatgggccg gtctctcttc 54780
ggtgattctt attgcttagg ttatcgcgca gttttaaaag ttgtcaaaaa taatcgttaa 54840
agtactttac aagggggctt atagtttgat aaagtaagcc ccagataaag agctttaccg 54900
ctaaatcctt aaactaggag attcaaaaat gaaatgcttt cacggtacta ctcaagaaaa 54960
ctttatcaac ctgattaata acggtgataa gccatcaggt gcatggaatt gttcggatat 55020
ggacggcaat ttttatgttt atccagaaaa taaaatctat ggcgacgatg cggaagagat 55080
aaaatcggaa ggaattcaac aagcgctagg aaatgccact atatcagcgg cctttcaaat 55140
gaaaactcaa aacattgtta tcttagaact tgatattcca gaaaatgagt taaacgatga 55200
tttctcttgc gataatatgt ctagcgtggc aagttttaca gagtattttg atataaactg 55260
gattaaaaaa gtttacatca ctgaatttaa tgctatgttt tcgccttttt gtcttccttc 55320
actggataac ccgaatttaa actatattga ggagcctcta gaacttctcg ctaaaagtat 55380
ccaacaatca gacagtattc aggttttttg tgacatcatg gacacactga caggaaacat 55440
tacagaaaga gatttaaaga gctttttcta ataatctata caatcacctt ataaacacta 55500
tttagcccct ataaggggct tttagggggc ttaaatcatg cttataaccg tttatcttat 55560
cctttctatt tgtaatgggt attcatgcaa ttttaaaggg cttgaggagt ttacaggaag 55620
taaagaaaat gctatacagg tttgtcagat agcaagacaa gactaccccg ccagtgatga 55680
tatacaatgt tactttaaga cagaagacaa ggacggcgtt tattttgata gtgttgacgg 55740
ccaatatgag attatcattg aaaaagacta gacaagcctg ataaaagtct gtaaattgtc 55800
aatcaacggg agggataacc tccccactaa gtaaagtcct taaactagga gattcaaaaa 55860
tggcttatgt aaccgtaatt accgacaaag ctggcgcatc ttggtcaact caagtaagtg 55920
ataagatgac acctatgcag tgcctaaaat actttgaaca atggaacaaa ggcgagggta 55980
caagcccctt tcatgtgatg cgtatcgttc acactgacaa ggaagggaat aaaacggcct 56040
tgaatagtga atactacgca accagattcg aaacgagaag taaaataatg aagctattac 56100
gtgaatccgg ttattctcat atcgccgctt taatctggga tgatttgctc aaaggccaaa 56160
agatgaacta tgtaaaacct gaaaaaatat tcatcagtta atcaataact tacaaaaaac 56220
ttttaaaaaa gtgttgacaa cccctcttga cattggtaac tttgttcgag ggggtttatc 56280
taaaagggat tcacttaaaa gattctttat agataagctt aaaaaattgc ttgcaaaggt 56340
aagtcaaaac aagtaaatta ctaatcaacg ggaggggata accttcccca ctaagtaaag 56400
tccttaaact aggagattca aagatgaaca agttcaaagc tatcaattac attcgttcaa 56460
gtgctgtaat gtcaaaacct gttaaagaca cttacgaatt tcgctgtaat ggtgtacact 56520
ttgcaaccat caccaagtca gaaaatgggg cttactacgt tcaccgtcgt aatgttgcaa 56580
ccgttgtagt ctctcacttt atgcaagctg tcgcggaact tctcccgatg tttttagggc 56640
tttacttaga tgagtgtaaa aacgttcaca aacacgttaa aaacctttta aacagttata 56700
gaatggctta tgagcgttct atcaagtcct taaacaagtt ttataaagcc cctgtaaaca 56760
ctgtttcaat ggcctatgct gcaaatggtg agatggtaaa cttatccaat gtggaagaaa 56820
tcgaactgca taaccagcgg cctgaaggtt actctaaaac catcggtgaa ggcttatctt 56880
tcaatatcaa gaatttagaa aaaactttaa aagatattga taaagatatc gacgagacaa 56940
tcaaaaatct gtaaattgtc aatcaacggg gctaaaatag ccccagatac cttcaaagct 57000
ccttttcata ggggcttttt tacgtcttat caaatccctt tccctttcca gaatcttacc 57060
cattgattcg atagaatcta tctcatacct taaaaagcaa taactctata tcttactcta 57120
tagcctttta ttaaatcaac ttaaatataa gcccataacc cgagagggaa gggcaaccat 57180
ttaaacgctc tataaagctc tataacagcc ttattaacct gacttaaggg attgcattaa 57240
ctcaccatta aaagcccttt ataaagcttt atagaggctt aaaagagcct taaaagaatc 57300
ttataagggt gtaagcttta tcatcttgtt agctatcttg tgaataacta aaaagggctt 57360
gataatactt aaaaggtatg ctatggaaag cttaaaagtg gctataaagt tacgtaaaag 57420
ttgcttaaaa gttacgtaaa tgctagcccc cttaacaatt tcttaacaat ttcttaacaa 57480
accctacata gttatgcttt taaggcactt gttagcccct tgttagcccc ttgttaaatc 57540
tctgttaaga ccacgtaaaa gccttgtaag agtctttggc atagccttaa aagctcctgt 57600
aaagccctat aaagagctat ccatacagtt gaggggtaaa tactagggag agacttaaaa 57660
gagcctataa agagctatag agagctttta agaggtatgt ttaaggaatc actaagaaag 57720
gaaagggaac gataagaaag aagagttgtc atgttaatag cttgttaaaa agattaaaca 57780
cttgttagga acttgttaat tatcttaaca aaataactct ttaaaattaa cttgttagtc 57840
atctgttaag catctgtaaa gcctattcag cacttgttag aaaactgtta cagccttgtt 57900
agttctgtta agaaactgtt aagaccttta aagagttgtt aagaagatgt gagaactgtt 57960
aagagattgt taaggatatt ttaaaattct gcaaaggggg tgttaagggg ctgttaaggg 58020
ctaccaagca gatatacatc ctaatcagat atttttcagt tccttcaaag acactaaaaa 58080
atgacttgat agtcctattt tataaaaatt ttaaaattct gaataattac ctatacagtc 58140
ttttaaaaat acaggtagaa acaggcttat aagatacctt gttagtaccg tgttagacct 58200
tgtgagataa ggtctgtaaa gatatccctg ttaagtttgt taaaacagtc tatatagaag 58260
cttagtaata cctagtgaac acctatagag ggtatcatag ctaacataat gggttaagag 58320
gtgtatagga gcttctaaga gggtatagag aggtggttaa ggtgattgta taggttgtta 58380
agaaggcttg ttagaagagt gtttagaagg cttgttagag ggctagttag aagggttata 58440
cagagggcta ttaagaccac tagatagtta ctaagatact atatagtact atataactat 58500
atagagagta agattaagaa ggttgttaag aaggatagat attttgttca catatgatgg 58560
tgacccagag gttgggacaa ccgataaaat tttataaaaa taaaaagccc ccaattaagg 58620
aggctcttaa gatattttaa atcagtgttt ccagaactgt ttttaatgat gacttggttg 58680
tctgataaaa atgagtgaat ccattgttgt tatcaaatat ctgcatcatg atatcatctc 58740
ccttgattgg gtacactcgg atatcatagt gaaacccatt aaagacagca gaaccattac 58800
agaacacact aaaatcatcc actgagatat actcttcatg acctgtgatt tttatacaga 58860
cacctttact tggtctctta gggttgatat cttccatcac agacatactc tgtccaccaa 58920
gagataccat agtgaccact ttacggttct tctgtttaaa gtacatttta actttctgta 58980
aagtgtcttc atagaacgga taagacttct ctacaaagat tccttcaagt ggctctttac 59040
cattcttaac ttctctgata tgccgtactg agaagacaag ctgctctagt atgtcaaggc 59100
tttctttgga taatgcttct cgaacatcat tccagtcaag gactaggtag ttaccttcaa 59160
cttcttttaa gttatctttt tccatagcta tcacaccatt atcgtaaaac ttggtaatct 59220
ggtactgcaa acttgttgtc attgttctta atgaggtctt cttctgttac taaaaatgta 59280
agaccacaca agtgtgtata gcggctatca tactcaaata ttttaagtaa gtatttacca 59340
tcatgataga cagattctac tacacctttt gcaacaatct ttccagtctt gaaatcttta 59400
agagggtatg tattaccaac tttaggaact gttttcatct caggttcttt tagcttctca 59460
ccaacttcta catcacctag taagttagaa ctttcaaaag ctatatgttt catataagct 59520
tctgagcatt ccaagataaa gcaatgcaca agagccttag cagcatgtct tgaacaattc 59580
aacttatcac aaatctcttt gtacatttta tctttagttg agttgtcaag cattgctttt 59640
tgaacgactt ttttaatttt catattcgct accttctaag tttctacaaa ggggctatca 59700
agtaacccct caagttacta ctcagacttt tcagtcttct tatcggattt cttactagtc 59760
ttcttctcag ctttctgctc aggttccttc tcagaagttg ccttgataac catatcaaga 59820
agctctgata agtcactctt cagcaaagta acaggctgct cgttgtagaa gtttttgtca 59880
agatagactt taaactcttc tatagaaggg aatccaaaaa ttggtacagg tgtcttaatc 59940
attacttagt ctctccatag gcatctttta agattgcctt gttaatcatt tcagctattg 60000
caaaacgaac acatacagct ataattgtag gttgcattct ttacttacct cttcaagttt 60060
aacctcagag tccttaagtc gtttcttatg aactctgaac ctcattagtg agatgatgat 60120
accagctaaa ggaacagcat acaagtacca aatatcttgt acctttgcgg taatctgtgt 60180
aattgtcaaa tcttgagctg tgtagctgat tactgatact gccagtaacg ttgctaagaa 60240
gaatgccagt cccataacaa tacttaggca gcctctggca tggtgttcat gcttaatgca 60300
cttattcttt tcgtcaacaa ctctgttgta gtcatttgca aagtctctgt agaaagcttt 60360
caggtattct tgagtgtagc cttcaaagca acctgtgcca taccagtaat caccaccaaa 60420
atgtgctatg ataccggtat ctccatttgg tttcgtcact cttaaagaag tgtgatggcg 60480
agtatcctga aaatctatct tgtatctttc ttcgtaaatc attttccaac ccctctcaat 60540
cgtttgttcc attcaaggtc aacaaatctt tgcagaactt tccctctagg tgttacgcct 60600
tttaaatctg catcatagca atggtaaaaa tcaccttcag cgtgcttacc atagcatttt 60660
ctgatattct ggcttctgcg atactgttct aacagggaaa gtttgtagta gtcttcaact 60720
gtagttagtt ttttcattct ttcaccatgt ctcttatcta gaaactaaaa agggaactac 60780
ctctcgatag ctcccatcat aatcagtttt gtatactcgg tcaagagtat tttttaggca 60840
gtaatgcact gcgcttgtta ctctcttcac tcaatagtgc gataaaatca tcacttttac 60900
ctttccacgg gctgaatgaa gggatatgtt cacggatagc ctcaatcact gttttaagtg 60960
ctgtgttctt catccaccaa tcagagtctg ctgatgctac cacgtagtat ccaccatcat 61020
cagttaaaat cggctctgaa atacatggtg aaatcaaagt ggtgatgatt gaaccattgt 61080
tcatgtagta tgtacgcttc ataacctttt caagaagctt gtgaaggtca ccaaacagtg 61140
ttacaagttc tttgtggtca atatcttttt cgaggtttgc tcggtacttc aatagtgcag 61200
catcaacctc tctgtcgtta aacttgaaat attgaacttc atcaacaagg caaacactga 61260
tgttgttaac attcatagtc agcttgcaaa cttttgcagg agagttgatg cttggttgaa 61320
aaagtgcaac acacccttca atatttccaa agtgttgtac aagtgttact tgtggaatga 61380
tatcctgttg ctcttcggac attacagttt cctcattaaa tattttttaa gttgtgtgtt 61440
aggtttctca aaaacttcta tagaagatat cttgacagct tcaccacgca tacctgtatg 61500
tttgaatatg taatactcta acatccctat gtactttgct ggtaagtcat atttgacata 61560
caccgtcaca gaatcttcaa attcaacgac cttcgtacca tctttgtagt taaactcttt 61620
aggatggttg tagttgtcta tgtcatacat taaaaagagc cttttacgtt gtaagtcaag 61680
cttatttgtt aaagactctt taatatctga gaagtttttg tacactgtga catggagctt 61740
actcatcgtt aaactctctg acgaagaaat agttgacttt gttattgcct accataaggt 61800
caaagcaggt attctcaaaa ggtttgttgt aggcatcata ggaagtgtgt gtatcagcta 61860
cttgtttagt attgagcttt ttacttgatg ggtttaagag gccaccatac agtttcttga 61920
tagcctcttc tacaacagac cagataattt cgtcagtaat ttcttcgtac aggtcaagtt 61980
cacgctctag tgagaagtca caaataggga aaacaacctg tactttactc attaacgaat 62040
aacctgtgag acttgaattg cttcggttac tgacaagtca cctactttct cttctttgat 62100
gaaggtaaag gtcagttcaa caggctcttg ctcttcttgc tcaactggaa caccgttaag 62160
ctcatcaaag attgattgag tagtccagtc ttctccacca tctggatagt ggattgcata 62220
aagagggaac tctatatcaa accaaccaaa tacagggcaa ccatcttcat catctcgtac 62280
tcgatggagt gtgcttccag accctgtaaa ctgaaggaat aaatctccgt tgcacttgta 62340
tattactgca tcggcaaaac ctatgtcttc tcgaccatca tggccaaacg tgtcaacttt 62400
tacttggata tctctaacat ctttattcat aaaaacctct ctccgcttgt tcgatttttg 62460
ctcttaaagt ttcgcaatgc tgcaaatggt agttagcttc ttgttcatgt gttgctaact 62520
gcctttccag aacagattta tagtaacctg caagttgctc ttttgtcaat gggtgctctg 62580
aaataatatc aagtccactt tcgccaagag ggttgaattg tccatgctca ttgacattgt 62640
aagtctcaaa cttgaacaga ccattgttca cagactcaat aggtgacagt tcaacaagct 62700
ctacagtaaa ccgtgtatac accttgtcac ctcttgaaac tacattgttt ccaacaatgc 62760
gtacaagcga gccattacga gtgtatttaa aagtcccaaa gtcactagcc caaagtttca 62820
tagttttctc cttagaaaat aaaaaaggct cccgaaggag cctatgaaga tattactttg 62880
gatatacact gtcaagataa atgtcagctt ccattctacg tctgtttttc aggccgtttg 62940
aggtaacctt cttacctttg actgtaacct tgttccacca ctgcatagct tctgcacaac 63000
cgaccttatc accagcgttg tggcgcttga taaatgtaga gtcctgcata gccgtgatgc 63060
caatgttgta tgtttcactt acaagtgcgt caaactcatt ctgagaagtt ggaaccttga 63120
tgactttgtt cactgctgca acgaactttt caacatctgc gagaagatac tgttcagctt 63180
gttcagcggt aatcttcata cccatcttaa caggtttgcc agaaatacgg attgttccgt 63240
acccaattgt tgggattcca gcagaatctt cgtaagcctc tagcttaaga ccttcaaaga 63300
acttaatagc ttctaaacct tttcttgaga gttgcattat gcctcccctg ctgggttaac 63360
agtaactgtt gctgcattag atgttacaga gccacctgca ccagtaacta cacatgtata 63420
gcttcctgca tcagcagcag ttgcaccttc tttggtgtaa gttgcagtag ttgcatcgag 63480
gatgtcttcc tcacctttct tccactgata acctgttgca cctgttgcag tcacttccaa 63540
gttgatgttt tcaccttcat tgatggtctg gctagaaggt tgctgtgtaa tcacaggcag 63600
tgcgttgaca gtaactgttg ccgcatctga ggtaacactt gtcccagcct ctccagacac 63660
aacacaagta taagaaccag catcagaagg tgctacagac tgttttgtgt aggttgcaga 63720
tgtggcacta gtgatatcct caccaccctt cttccattga tatccagtgg cgttggttgc 63780
aataacgctt agggtaagtg tgccaccttc attcactgtt tgattggttg gctgctgagt 63840
gattgctggt ggcttgatag catccttcaa cttagcattc aacattgaga acggcttaac 63900
ccttgccagc cattcacaat aaattgtatc aacgtcttta ccattgataa ttgcatattg 63960
taagtccatg aagaagtcag aagttctcat ttgagcaccg atgctgtaaa gcaactcatc 64020
actaaatggg actttgtagt ctggtttgta gtcaaactcc tcaacttctg caatatcagc 64080
ttcaggccaa tacgaactat aggtaagcgg taaaatagcc tcttgatatg ttctaaaatc 64140
atatctctta ccagctttta cgtttgcaat gaagaccttc acaaactctt tgaagtctgg 64200
gtaagtcttt gctttaatca tttcttttta gcctctaaat gtgcgtttag tctaaaataa 64260
cctttgtgat gcggagatac atcaccaaca acaatacctt taaccttata gcctttacca 64320
attacatact ggatatccat aaggaatctt ccaagaccaa caatatggaa cattggatat 64380
aatagattct cagaatctgg aatattgatt tcaggtttgt aggtgcgttg ttcgcttgtc 64440
ggtgtaatat cttcttcagg ccatttactg agtctccatg tcaaaggtgt tgggataaca 64500
tcattaaggt tgtacctttc accattttgc acggaattga tataggattg taggaagcaa 64560
taaaagtctt ttgcactctt agctttcatt aaatatcctt ataaatagtt aaaggggcat 64620
cagcccctat tcacacttaa gctttggtct taagtaggtt cttaagtttt ttaagattca 64680
tgctctcatc aaacttaaaa ccaaatccat tgacataatc aatgaactcg gatttcttac 64740
tgagtgacaa tgcgtaactc atatcaaagt tcttacttgt ttctgcgtct ggactagagt 64800
tcacatcaaa tgaaatttct gtaggctctt gcttactttt tctcatatta tcaattaaca 64860
aacatgtata gtttgctgga tagaagttta cactaccatc aacgtagctg tacttattat 64920
tatggataag gttggaaaat accttaagga atgtttctga gtcgtggcaa acaagtttga 64980
attggctgct ctctccaagc gcaactaatg agtacggact ttcactatca aagtcatagt 65040
ttggatttga tgccgtcact gttacacgca tcaggttatt agcaatagtc ttctgtttaa 65100
aagaattttg aacatagtga agacccctct cacactgatt gactagctct ttaagtaggt 65160
catttgcata tggtgagtta attgatacag agccgtcact attgtaatta aaacttactg 65220
ttaacatttt tcagttctcc tgcgatttat aattttgctg caatagagaa acgaagtaaa 65280
ctcactggaa gaaaccacta agtgttgttc actagattga gccattgccc tgaaatacat 65340
tccagtcaaa gcatcatagt gactattctt tccaattgag gttatatagc catgactctt 65400
tgcagcttga tataaatcta taatatctgc gatttctaat tgggaactac acatagataa 65460
tcctcttgtc ttctgtaatt ttgaaagaac aggtacagac caatttcaga ggacgaatag 65520
tagattaatt ctaccctttt tgattcctta cagtgattgg tattcttacc tttcttcggt 65580
ttatttgatt ttctcattaa gtctcctttg tagaacaagg cagacctaca gctttaagta 65640
agatggtaca gtaatccata caacaagtca agaactattg caagttaaaa aagttgttga 65700
caaagcactt gacaaggtat aggcttataa atacttagaa gataacttaa aaggttcctc 65760
taaaaaggct atttaaatag atatctttct ggatatctat cttttatctt aaaagccttt 65820
taagaggtct tataagaggc ataccaatga aaaacagaac aaacaaaggc cagtttaaaa 65880
aaggccagtc tggaaaccca tctggaagac caaaaggctc tcgtaacaaa agctcactcg 65940
taaaagctca actgaccatt gataattctg ctgagtttgc tgcaaagttg tttgaagcga 66000
ttgttacaag ggacgctgct aagcttgcag agttcgggtt aaacacagac gacgtaacca 66060
tcaagagcat gattgaggtc ggtaaaacta tcatgaccca ctcagcaggt gagatgaaag 66120
caattgcagc agatactaag aagactcctg ataacagtag tcagtctcag acagaaaaca 66180
aaccaacgtt ctctgctgtt gcaactctca aaaaataatt ttaaaagatg ttgacaggct 66240
ctacaaagtt actctaaagt ctgtcacatc aaacaacaaa tgagagaaga gagtaaaata 66300
tgagcgaatt atttaaacat gcgcaccttc atgcaggtcg aactgaaaat ggtgctgtaa 66360
accatacttc atcaatgtct gctcttgtag acttttacaa agctgctggt tcaagccgta 66420
gcaacgtaga aatcttacca gacctgttct acaaagcttt gcgtgaggat gttgatgttg 66480
cagttcgtat tttactgcat atgcgagatg tacgagaagg tattggtgag cgtaaagcct 66540
tcagaactgt tttgcttcaa gcaattgaag ataaagtttt agagcctaca caggttcttc 66600
gcattatgga taagattgca gaacttggtc gttttgatga cttcaaaatc ttcgtaggta 66660
ctcgtttcga gacagatgcc ttcaaacatt tagaagcagc attactagac cctgcaacag 66720
caggtttggc agccaagtgg ctaccacgag tcaaaccacg tcataagcag tttgtaaaac 66780
gtttctgtaa gtttgccaac ttgagtgaga aagagtatcg cacactgttg tctgcgctct 66840
ctgatacagt tgagcaaaaa atctctgcta atgagtttgg caagattgac tacagtaaga 66900
ttccttcact tgctgctgca cgttaccaaa aactgtttaa ccgcaaagat ggagaacgct 66960
acaaagctta catcgagtcc ttatcaaagg gtgagactaa gattaatgct ggtgctgttt 67020
acccatacga tgtgattaag tctgttaagt atggtaatgc agatgttgct aatgagcagt 67080
ggaaagcact accaaactgg atggcagagg gtgaaaacat tctctgtatg tctgatgttt 67140
cacgttcaat gtcttgggtg aattttggct ccattactgc tctggatatt ggtgtatcac 67200
ttgcattgta tgtagcagaa cgcaatacag gttgctttaa aaatgagtta atggtttact 67260
caacaaaccc tcacttcatc gaactgagtg gtgatttacg aaaccgtcat cgtcaggtga 67320
tgcggcacgt tgaatatggc tcaactaact tacaagcagc ttttgaccgt attcttgaga 67380
caggtaaaag aaataacttg actcagaaag atatgccaag taagcttatc atcttctcag 67440
atatggagtt caatcaggtt gatggagcta acggtcgcac aaactttgaa gcaatccaga 67500
gtaagtacaa gaaagctgga tacgaaatgc cacaactggt attctggtac ttagcaaacc 67560
gtaatggtac ttgcgaagta tctgttaagg ataacggtgt agcaatggta tctggtttct 67620
ctccagccac tttaaaagct ctgcttggtg gtgagaagtt tgaccctatc agcgtaatgc 67680
tcaaagcagt aatgattgac cgttatatct ggtaaaaagt tttaaaaagg gtattgacaa 67740
tgtgtttgat accctttaat atgttctaca tagaaacgaa acgagagctt ttctaagata 67800
ctgaaaaata ttttaaaaaa gttcttgaca atcactaaaa aataatgtta aagtggttac 67860
atagagtttg aaaagtttat ctctgtttag ctcagcttgg tagagcgttc cgtttggggc 67920
ggtaaggccg gaggttcaag tcctccaaca gagaccaaat taatgttcca gtagacaaaa 67980
tggtatagtc accactcttt caaagtggat atttgagggt tcaaatccct tctggaacgc 68040
cagttttgac agaagaccaa ttacagcaaa cttaatctta ttcatctgaa ggtaaatcgg 68100
acaaagaaga gatttggtct ggtcattaag aattgcgggt atagagaaag ggcgtctcac 68160
atgtctcatt agcatggtat cggcaggttc gactcctgca cccgcctcca aatttacaga 68220
agaccgttta cagcaatatt tattaaattt ttagcggaaa aattgaaaga tgtggtctgg 68280
taatactaaa catggttagg aagcacataa ggtatgtgcg gtcgcctgtt aagcgaatgg 68340
cacagggttc gaatccctga ctaaccgcca aatttaggag agaagcgatt gctcaccgat 68400
ttgtaaggtg caagtcctta gcctgaatag aaattaaatg tgtcgttatc ccgtagatgg 68460
tagcggtggg gactgtaaat cccttgtcat tgagactcgg taggttcgac tcctacacgg 68520
cacaccaaca aaggttatgt agtttaattt ggttaaaata ctcccctgtc acgggagatg 68580
atgagggttc aaatcccttc gtaaccgcca attttgagag ggctatttag cccctccctt 68640
aaagagttct tacgagtatt ctttaaagga gcagaagacc aaatacagca agttacttaa 68700
aatttcaagc caatttaatt ttgaaaaatt taaaacttgg tctggctcaa caaatttaca 68760
gaagaccgtt tacagcaaaa cttaaacaat ctatttctcg gtaaagaaaa ggagaaggtt 68820
cgattccttc actcggcagg tgtcgagttg gtgtaatggt agcacttaag atgataaaca 68880
acggtctggt aattatcttt gtagataagt tattaagctt acctgtgagt atgcgaccac 68940
aaagggtgaa agggtaaatt ctagatttaa acccaaaaag gcacatcgct acagacctta 69000
agcaagtctg ggtagcttat ctacaaagat aatttaatgg aagtgtagca gaatggtgat 69060
gcggcagact tttaatctga caggcgatgg gttcgaatcc ctccacttct accaatatgg 69120
ttcagtcgca gataaggtaa tgcaagggtc tcataagccc tatgaatgtg ggttcgattc 69180
ccatctgaac ctccaattgt gggttagcat aaatggtaat gcaaacggct gataaccgtt 69240
agaagagggt tcgataccct cacctacaac caatttcaat aaagttgttg acattgagat 69300
gtgactactt tatagtaact aaaatacttc agaatatgta taattcaaac accaatacat 69360
gagcgttaga cgtaacacaa tgtggggaaa tttgttgtac atattctgaa tagttttcgt 69420
tgcgtagcgt ctattttgca aatttaaaat aaatgcaaac gacaatgttt ttctggcagt 69480
agcttgatag gctaaacacc agtgaggtct tccaatccct catcaaagaa tttggcgtac 69540
tagcccacgg tatgattaat aaggtgggca tctttaaggg ctttctaaga gagtctttaa 69600
agataattac ggggcgtatc tcagcggtct tctaaaccgc cgttaaccga gtaagtggag 69660
tatgcgggtt cgagtcctgc cgcctcgacc aattagttag cagattactt agacgaccta 69720
agcgggtctt cctgtaggga ggtggtctgt atctcatgtt ttccagaaca tgtaaataat 69780
ctgagaaggg gctttacagt gttgagataa tacggtaagc ttcgaattat agtatgggac 69840
gatgttaaga ctctaaggca tgagcaacgg cttccaacac cgtttcaaag gggttcaact 69900
cctccgtccc atgccaaagt cttattaggg gtaggtagcg gctaatggta gccaaactgt 69960
cttgaaaaca gttgccactg tagagatacg gtgagggttc gactccttta cttaccgcct 70020
aataagattt aaagccaagc ttcattcgga tgtcgctttg gtatcccgcg tgtattgtcg 70080
tacacactga taaaacacct agtaggtgtc acggagaaga gataatatca agctctccaa 70140
aggttctagt caccggatta aacaagacta tgcaaagacc tttttaggtc tttttagagg 70200
gcttctaagg ttattacagc ttgttgtgga accttgttgc ttaggttctt agaagttctc 70260
taaaaagatt taatgggaga ttgacggtaa ttggtaaacc taccatcctt agaagttggt 70320
gtttgagggt tcgaatccct tgtctcctac caaattaatg caggtgtagc aaaatggtta 70380
tgcggctgac tcttaatcag taagacgatg ggttcaattc cctccacctg taccagacat 70440
ttgggtcata gtttatatgg ttaaaattcg agttttgcaa acttgggaac tgagttcaat 70500
tctcagtgac tccaccaatt agtgcatcca tagtttaaac gggaaaatta cagtcttcca 70560
aactgaggtt gagggttcga ttccctctgg atgctccaaa caatgctgct ttcgtataat 70620
tggctattac acatcccttg taaggatgga aatgcaggtt cgagtcctgt gagcagcacc 70680
aattcagagg tcaagtgaaa gaccgctggt gtcaactgaa gaccgtaaca aattccacgg 70740
agttgagtta gcggcacaac ttcagacctc tttcacactc gcttagttta tatggtaaaa 70800
catcaccctt acaagatgaa gaaaaaggtt caagtccttt agtgagtacc atgttccagt 70860
atcccaattg gcagaggatg caagctcaaa ccttgtatta gtgacggttc gaatccgtct 70920
tggaacacca atttaaagag gctatttaat gagatatttg aagtatgctt catggatttt 70980
cttagcttta ctagacccaa ttgcagcaat cctagcagtt atcttagcac ctttcgtagt 71040
tccattctac agtgagaaga aaggacatct accttttgga ttcagatgga tggagaccta 71100
tgacaacccg attgatggtg acgaaggtca cgttaaaaga tgggctaaga ttagaaagat 71160
tggtaagctg ggtgtctaca tgcagagagt tgggtggctc tggagaaaca aagcttataa 71220
cttctcttac catgtattag gaagagatgt aaaagatgtt actaagtgga gaggtaatat 71280
caacgtaagt tctgaccctg aagataatca gactggttat ctcctaatgt ggaacaacaa 71340
tgcttgggga ttattcgctt ttattccgtc aattaaagtc tttggtaaac agttttactg 71400
gagaatttat gttggatgga aactaaaaag tgttgtccca gaagaaagag cattctcaag 71460
ggaaagagtt atgttggcat tctttattca tccactgaga aagtaaagat ttaaagggga 71520
ttagtttaca aggttaaaac ctcggtcttt gaaatcgaag aagttggttc aattccaaca 71580
tcccccgcca atgctccatt actccaattg gcagagaggc cagacttaaa atctgtgtta 71640
tgtatcggtt cgaatccgat atggagtacc aattttagcg gtatagcata actggcaatg 71700
cagcagtctc tgaagctgtc ctattaaggt tcaaatcctt atgccgctgc cacttctaag 71760
ggttcttacg agagtcctta aatgtggcct tatcataaac ggtaatgacc catgctgtga 71820
acatggtcta tacgggttca aatcccgtag gtcaccccaa tttatagtcc aagtagctta 71880
tatggttaaa gcgcgtgtct gaaaaacatg agaagagggt tcaaatccca ctggactacc 71940
aatttcaaag gtgcttaatg aaagagatga caggacaagg taaggagatt tttaatctct 72000
taaaaaccgg taaagggttt tctaacccac ttattactgg tgcagcagtt ctcggtggaa 72060
ccgtagctgc ctctacatca cttgtaagct ctattagctc tgtaacagac ccgacaatga 72120
atgagaaact tgttgctgct ggacttacaa cagtccttct aaacagcttt acaacaagcc 72180
tgacaagcac tacagcaact actaagaccc taacagacta tggtcaaaag tctattgatg 72240
agttttcatc acgtatgcag gtagcaaagg ggtattccaa tgttatgggt gcagcaggag 72300
aacaagttgg ttgtacacca ttcagtggta ttatgggtgt tgctacagag tatggtcaaa 72360
aagctattga catgattaac agcacactag acagtgttaa cggtgtgtta agtgacttac 72420
aagacgctat tgataagggt cttgatactg tttctgattt agctaatcaa gctgttagca 72480
agattaatga aggtatctca aagattacag cttatgcaga tgaagttgtg cagatgattg 72540
aagaagaagc tgcccttatt gcagagtacc ttaaaacgaa tatcaatggg ttcttagcag 72600
gtatcttacc agactggttt gatgatgctc gcaaaactgg tgtaatcgac actattgcaa 72660
caccagaaat gaagaacgca ttacagaaat aatggaagat taaccctaaa aggtaaggga 72720
gcagtttgct aaactgccag tagctgagaa atcggtgtac cagttcaagt ctggtatctt 72780
cctccaatct gaatccgtga cagaaatgga tatgtgcctg tctgcaaaac aggtttataa 72840
gggttcaagt cccttcggat tctccaagtt attttactct ctctcaatta aaataataat 72900
aatgccctgc tataagtatt cttcctttcg ctaccgaaga gtatttttag acagggcttt 72960
tttacagagt tatactttat aggaagactt atacagtatg tctctgacgt acattgtggt 73020
ttctccttga cggtctctgt gccgtctttt ttaaggggaa acaagattta ttatttggag 73080
aaaacataat gagcgagaac gtttataaag aactttatga agctaacaag aagttagagt 73140
ttatgcagaa tactatcatg gcaattgcag acagattatc tgtagcaaca ggcattgata 73200
ttaaagaagc atctatggat gcacttcttg atgctgttga tgcaaagttc gaagttaaga 73260
aagaagagac tgcgacagac tctgaataat tctatttgcg gaggcgctta atggacttaa 73320
atgctgttaa gcagaagcga gtggaagatg ttaggaaagt cctagctgga gagttggggc 73380
tttctgatga agtaaaagaa attatcaaat cattcggtaa agacccctct aaattccttc 73440
caactcaaat tctgacttta ttaagataca caccagacca agttcgcctt atctttaaat 73500
tgatgactga taagaactat gtagcccctc agccgggttc tcaagaggtc tttttaaaca 73560
ctaatgctga cttggttctc tatggtggtg ctgctggtgc tggtaagact gctgcattgt 73620
taatggactc tttaagattt attgaagacc ctaactataa tgctgtgtat ttccgtcgaa 73680
acacaacaca gttacaaggt ggtttatggc ctgctgcaaa gaaactattt ggtaagtttg 73740
gtgggattcc tcacgagcag aaaatgacta tcacattccc ttctggggcg actatcaagt 73800
ttacctatct agaacttgaa aagcacgctg aaggtcatca gggtattgaa tactcagcta 73860
tttactttga cgaaggtaca cacttctctg cttcacagat ttcataccta caaacccgtc 73920
tacgttctgg tgctgaaggt gattcataca tgaagatttc catgaaccca gacagagacc 73980
actttattta tgattgggta gagccattct tagatgaagg aggttatcca gaccctgaaa 74040
agtgtggtcg tattcgttgg tatgtaatga atgatggtgt gatggtttct gattgggaga 74100
gagacaagat tcttgaaatg ttccctcttg agattcctca aacatacacc tttatctctg 74160
gtacgattga tgataaccca atccttgact tcttagaacc aaagtatcgc ggtaagcttg 74220
aaaacaacac tcctgtgaac gttgcaagac ttcgtttcgg taactggaag gctcgtgcag 74280
aaggttcaaa ctattggcaa agacaatggt gtgaaattgt tgattcactc ccagaagatg 74340
tatttgatgt tagagcatgg gacttagcag caactttacc atctgagatt aaccctaatc 74400
cagactggac agcaggtgtt aagatgggta aatctaaaaa agacggttgc tattatatca 74460
ttgatgtagt aagatttaga gatagaccct ctggagtcga aacacaaatt aatttgactg 74520
ctgaaagtga cggtaagcga actggtattt ttatccctca agacccaggc gctgctggta 74580
aatcctacgc gacatccctc atcaggaaac ttgccgagaa aggctatcgt gcaagagcta 74640
aaccaacaaa taaagataaa gttacccgct ttgcgggatt ttctgctgct tctgaagctg 74700
gacttgtaaa agttttgaga ggtagttgga acgaagctta tttccaagag cttgaaggct 74760
tttgtggtga tggcaaaact aaagatgacc aagtggatgc taccagtgat gctttcaaca 74820
gtcttaacga agttaaatta ttcaagccac catcaatggg tgctcacaca gacttagtga 74880
gaggaaaccc atatgagggg cttagacgtt gatagctagg tgagaagaat ggcagatatt 74940
acagaaacac aagaaagctt accaccattt agaatgggtg aagtaggttc tttgggtctg 75000
aaggttaaga atggaagaat ctatgaagaa cctcgtcagg cactaaggtt ccctgaaagt 75060
attaaaactt tccaattaat gatgcgtgac cctgctgtag cagcatctgt aaatattatt 75120
aagatgtttg tcagaaaagt caactggaga tttgtacctc caaagggaaa agagcaagac 75180
cctaaaatgc ttgaaagagc agacttcttt aattctttaa tggatgacat ggagcatgat 75240
tgggcagatt ttattaactc tgtaatgtca ttctgcactt atgggttctg tgttaacgag 75300
aaagtttata agaagcgtca gggtaagaaa ggaaagtacc agtcaaaatt tgatgatggt 75360
ctaattggat gggctaaatt accaatcaga aaccagtcaa cacttgataa gtggtatttt 75420
gacgaagact ttagaaaagt tactggtgtt aggcagaatc tgagaaatgt ttcacatatt 75480
gctggagcaa ttaatctcgg agaaaggcca ctaacaagaa aactcccacg agctaaattc 75540
atgctgttta agtatgatga tgaatatgga aacccagaag gtcgttcacc attacttaat 75600
gcctatgtac cgtggaagta taaagttcag attgaagagt acgaagccgt tggtgtttca 75660
agagacttgg taggtatgcc aaagattggt ttaccaccag attatctgga tgaaaatgca 75720
gaacctgaaa agaaagcttt cgtacaatac tgcaaaactg ttgttaacga tatgattgct 75780
aatgatagag caggtttaat ctggcctaga tatatcgacc cagatactaa agaggatatt 75840
tttgagttct cattagtttc tagacagggt gctaaagcat acgatacagg ttctattatc 75900
gacagatatt ccaagcagat tatgatggca tttatgtcag atgttcttgc tatgggtcag 75960
tcaaaatacg gttcattctc tcttgcagat tctaagacaa gcttattagc gatgtcagta 76020
gatatcctgc tgaagcaaat taagaacgta attaaccgtg atttagttgc acagacttac 76080
gctcttaata tgtgggatga tgaagaacat gtgcaaatca catatgatga tatcgaaact 76140
ccagaccttg aagcaattgg ttcttatatt cagaagactg ttgcagtagg tgctttggaa 76200
gttgacaaag agctatctaa caagcttaga gagcatattg gtcttcctcc tgctgatgag 76260
tctcaaccag tatctgaaaa gctttctcca aatagccaaa gccgttcagg agatggctat 76320
aagactgctg gagaaggtac tgcaaagaca ccttcagcaa aagacccaag cacagcaaat 76380
aaggcaaata aataatggct gaagttatct ctatttcaaa tgctacacgg gtgcattcgt 76440
atagaggtgt tcttatcatt actgacaagt tatctgtaga ggctggctca agggtcagcc 76500
tttcaggtta tgtaagcgat ggtggagcct ctgacgtttt cactatttgc aggttacttg 76560
atgcaccaat gagtggaaaa ccgtttattt caggaaattg tagtgaaatt gttaaaattc 76620
catttgacag ttcatgcctt ttgggtgtaa agttatataa ctgcgagaat aaacgtatca 76680
atgttaatag cattgaagcc gctttcatta ccctcgacac tgcatttcaa tctccaatga 76740
cagttaataa agaaacaaac agacttgaat acattttttc acaaaatgat tacaaagtac 76800
ttgtcaaagg caaagtatat gatatgattg taaatgtggt agatgaatct ggtaaccatt 76860
caacagtcct taaacaaaaa gtaaggttta attaatggga acattaacta ttgatggtaa 76920
gaacaaaatc ctcgccacgc taaccccaac gactattatt ttacacaatg tagacccaac 76980
ggcagaccct acagcaaata aggttactca gccagtggct atttattttt ctgaaccgga 77040
taatggctta attgcctcag aagacacagt taacattact gttccagctt ctgcaacggt 77100
ctcacactat agcttgtggg atgctaacga taaatgcgtg gcaactggtg ctcttagcaa 77160
acctcaattc tttgctgaag aaggtattta tgttatctct tcagtctctg tagatttaaa 77220
caaatagggt gagtaaaaat gagttcgaat atcttcagac ttgctgatag attattcaac 77280
caacctttac tagccactga atcattagct cactcagcag caacttatgt gaataacaga 77340
ttgctgggtg atgtccaagc agcagtaaac tttgataaac ccaaaggtga agcaagaagt 77400
cttttaaaag taaaagatga cattgccatt atccctatta tgggtggttt aacccatcgt 77460
atgacattca ttgatgcaat gtgtacaggt ggattaagct cttatgaggg tttacgtaga 77520
ggctttgacg aagctttagc agatgagtca attaagacta ttgttctgca catcgattct 77580
ggtggtggtg aagcttcagg ttgctttgaa ttagcacgtc acattatggc ttcaagaggc 77640
caaaagaaaa ttattgctta tgtagatgag ttcgcttgtt ccgctgcata cgctcttgca 77700
tcttctgctg aagaaattat tgcatcacca gatgcagatg ttggttctat tggtgtcatt 77760
atggttcatc aggaattaac taaagcattt gaaaagaatg gcgtgacaat taacgtcatc 77820
aaagctggtg agtttaaagg tatgggttca ccattccaag cactttcaga agaaagcaaa 77880
gaaagacttc aaaagagaat taatgatacc tacgcaacct ttacaggttt tgtagctgaa 77940
tctcgtaatc tctctgaaga agctgtaaag aatactgagg cgaatgttta ttctgctcag 78000
gaagctcttg aacttggttt aattaactca atcatgtctc aagatgattt cttaaattac 78060
ttacaaggtt ctgaagaggc tcctgtaagt ttaaacgtta acaattcagg tgaagaaatg 78120
actgaacaag aaaagcaaga actagaagct ttgcgtcttc aggttgctca aatgaaagct 78180
aaagaacagg aagctgcttt gtcagatttg actaataaga tttctgcttc tgctgaagct 78240
tttggatttg atgcaaaaga agctgcaacg actattttag gtgctggtct tgataaccct 78300
ctgagtgttc tgtttatgaa tgctatggaa ggtgctaacc agaaacttaa tgaaactatc 78360
gcgtcccatg cttctgcaat ggaagaaaaa gaatcagaaa ttaccaagct gaaagaaact 78420
gctggtgctg ttcttgaaca ttccaatgct atggaagaag tgggtaatga cggcgaagct 78480
gatttggttg aagaagaaaa agaaccagct aagaatgctt ccgaagacac cgctgaacaa 78540
cgcaaactgg ctctccagaa tgctctaaaa tctcttatca aataaggaac acaataatgg 78600
catatcaagg ttttactaag ttaggtaaca gagaacctct gaatgatatc attctttggg 78660
aacaggttac cccaacaggt cactctcgca aagagtacac tccggttgct tcaacagaat 78720
accgtgtagg tgaagttctg aaagcagatg gtactaaggt tacagcaggg caagaagctc 78780
aggctgattc agtatgtatc gttaactttt atgcagacct gcaactgtct tatcatggtc 78840
agttgaaagt tgttggtatc caccgtgatg cagaactgaa agacatgctg actcttgaag 78900
caagcgttga cgctgccgca gtcaagaaag ctctggctgc caaaggtatt gatttcgtac 78960
caactggcct gtaataacaa taataagaca ttctggagaa ttttacaatg ttgactaatt 79020
ctgaaaaaag cagatttttc cttgctgacc tgactggtga agtccagtct atcccaaata 79080
cttatgggta tatttccaac ttaggtctgt tccgttcagc accaatcacc caaactactt 79140
tcctgatgga cttgactgat tgggatgtta gcttgcttga tgcggtagac cgtgatagcc 79200
gtaaagcaga gactagcgca cctgagcgtg ttcgtcaaat cagcttccca atgatgtact 79260
tcaaagaagt tgaaagcatt actcctgatg agattcaggg tgtacgtcag ccaggcactg 79320
caaatgaact gactactgaa gctgtagtac gtgctaagaa gctgatgaag attcgtacca 79380
agttcgatat tactcgtgag ttcctgttta tgcaagctct gaagggtaaa gttgttgatg 79440
ctcgtggtac tctgtatgct gacctgtaca agcagttcga cgttgagaag aagactattt 79500
acttcgacct tgacaaccct aatgctgaca tcgacgcttc tattgaagaa ctgcgtatgc 79560
acatggaaga cgaagctaag actggcactg taatcaacgg tgaagaaatc cacgtagttg 79620
ttgaccgtgt attcttcagc aaactggtta agcatcctaa gattcgtgac gcttatcttg 79680
cacagcagac tccactggct tggcaacaga ttactggttc tctgagaact ggtggtgctg 79740
acggcgttca ggctcatatg aacactttct actacggtgg tgttaagttt gtccagtaca 79800
acggtaagtt caaagacaag cgcggtaagg ttcacactct ggtgagtatt gacagcgtag 79860
cagacactgt tggtgttggt catgccttcc ctaacgtatc tatgctgggt gaagctaaca 79920
acatcttcga agtggcatat ggtccatgtc ctaagatggg ttacgcaaat acacttggtc 79980
aggaactgta cgtatttgaa tacgaaaaag accgtgacga aggtattgac ttcgaagctc 80040
actcttacat gctgccatac tgtactcgtc ctcagttgct ggtagatgtt cgcgctgacg 80100
ctcaaggtgg ctaataaact taggagggtt atgaatgtgc tatacaggcg acccagccaa 80160
taaccctctt gatagagtaa gaatcctctg cacagacact aataatgatg aaattcttat 80220
tgagcagtct gtgctagagt ggttctatct agaatctgga aaggatgaaa agaaagcagc 80280
catcaaagct cttaaatatt tactctttca agtagccaag atgggagatg agaaggttgg 80340
tggtgtttat ttacgtaact cttccagatt caagtctctg aaagctgttt atgatgacct 80400
tgttaaaagc tctgtttcag gactacccta tgcaggtggt attaatcagt gtgatattga 80460
catgcgtcgt cagaatcctt gctctgtcaa gaaatacaca gaatatggtg atgctgccag 80520
atatgaaggc agagattact gcaaccgtgt taatggcgta tttattatcg agcgagatga 80580
ataatggtta aaagggttat tcaccccgct agagcaaaat tagtcggggc tatgaagaac 80640
ttgcaaacgg ctaatgctca agttgggtat tttcaagaac aaggtcaaca tagctctggt 80700
ttttcttatc ctgctttaat gtatttacaa gaagttattg gggttccctc agcttctggt 80760
aaagtatatc gtaggttgtt tgaaatcact atgatgctaa acaaacagac cttgttagag 80820
cagactaaga agaatctata taagcaactt agcagtctca acacagaccc ttcaaatacc 80880
ttagaagcat ttgcaaagaa tgctcaaaag gcaattaaaa gaggttttgg taactctgct 80940
atactccctc ctaacgcacc ttccacagtc aagaaaaaag gctttaacgc acctcttgtt 81000
gagacaggtg acttaagaga taaccttgct tataaaattt ctactaagaa gggtattaag 81060
aaatgagact cttaaacaga cacagctttg tagtaaagcg taaagtctct gaagacggtt 81120
attataatga tgatggtgat tgggtagctt cacaagatat tgtagaggtt aactgtaaag 81180
gtaatatcca gccatatatc aaaggttctg taaagaatgg tacacagatt gctttaccgg 81240
aaggtatcag acttaccgat acaagaatcc tgtatactac atataaactt agaacttcag 81300
atgatgtaga gtggaatgag tctgatattg ttatgattga tggtcatgag tatgaagtat 81360
ttatgactat ggattggtca cagcaattag cccatacgtc ccattatgaa tatatcatta 81420
ttagaaggga taaaatgaat gcagttagaa acagcagaac ttgaaaaagg tctagttaga 81480
accttagtgg atgttattgg tcacagatta gctcgtgata aaaataatag gccaaacgta 81540
attagagctt acccttctga taactcaaat gacaaaggtt taaaacctga ccagccattt 81600
atcactgtat attgtcaaga tgctgcaaca ccttatggtt gggttcttga taagtttgtt 81660
gaggatgatg tagtttgcta cagaattgct tttcagattc cagtattaat tacggtgaat 81720
ggtaaaggtg ctcacagtat tatgcttgag cttaaacaac gattagagat gagttcagtc 81780
agagatttaa tccttgaaga gacaggagct acagtactag acactggagc tatcccgaat 81840
gactatactt atctcaatac agatttcgaa aattctgcgc ctcttgttgt aactcttgta 81900
aaaaactcag tcctgaagga tgaacgtgga agtattattg agcgtgtcat tgttgatggt 81960
gagttggttt atgaagaagg acaagagcca ccagaatata ctatccatct agatgtagac 82020
tccaaagggg taaaataaat gtggaatcca attgttaatg tagatattac attgaacacc 82080
gcaggaacta caagagaagg ttttggtttg ccactattct tagcttcaac agataacttt 82140
gaagaaagag tgcgtggtta cacttcctta actgaagttg ctgaagattt cgatgagaat 82200
actgctgcat ataaggctgc taaacaactt tggagtcaga ctcctaaagt aactcagctt 82260
tatatcggta gacgcgctat gcagtacact gtatcaattc ctgatgccgt taccgaaagt 82320
acagactact caattactgt agctgctggc ggtggaattt ctcaaccata ccagtacaca 82380
gcacaaagtt cagacactgc tgagaacgtg ttacaacagt ttaaaacaca gattgaagct 82440
gacccaacaa tcaaagataa ggtttctgtg aatgtaactg gtagtaatgg ctctgtcaca 82500
atgattattg ccaaagctgg tgataatgac tttgtgaaag taacaaccac agcacagact 82560
gtatatattg caagtacaac tgctgataca gcatcaactg ctctggcagc tatcgaagct 82620
tactctaccg actggtattt cattgcagca gaagacagaa ctcaacagtt tgtcttagca 82680
atggcttctg agattcaggc tcgcaagaaa atcttcttta cagctaactc cgatgtaaca 82740
gccctgcaag gtacagaatt agccagtgca aatgatgttc cagcacagct tgctaagaat 82800
atgtacactc gtacagtttg cttgtggcat cacgcagcag cagaagatta cccagagatg 82860
gcatacattg cttatggtgc tccatatgat gcaggttcaa ttgcttgggg taatgctcag 82920
ctaactggtg tagcagcttc tctacaacca tctaatcaga gacctctgac aagtattcag 82980
aagtctgctt tagatgtacg tcactgtaac tttattgacc ttgatggtgg tgttccagtg 83040
gttcgtagag ggattacttc tggtggggaa tggattgata tcgtccgtgg tgttgactgg 83100
ttagaatcag acctgaaaac ttctctgaga gacttgctaa ttaaccagaa gggtggtaag 83160
attacttatg atgatactgg tattacccgt attcgtcaag tcattgaaac ctctctgcaa 83220
agagcagtca acagaaactt cctgtcatct tacacagtta atgttcctaa agcctctcaa 83280
gttgctttag cagacaagaa agctcgtatc ctgaaagatg ttaccttcgc aggtatctta 83340
gcaggggcta tcttggatgt tgacttgaaa ggtacagtgg cttacgaata atagaggtaa 83400
attggaatgg ctatgtatca gcaatattcc cctaaagacg ttgtatgtag ctggaatggt 83460
attgccattg aaggctttgc cccagactca ttcttgcgtc tacagagaac atcaccactt 83520
gtgacaccag ttgttggtgc aggtggtcag gttgctctga caagaaatgc agacaagaca 83580
ggtactattg agattgagct aatgcagact tctctctcta accagatgct ttctgcaatt 83640
caagctaaac aagacaatat ggaacttgaa gtagatatct cttctaactt cgtaatctac 83700
gacccatcag gctctgttct ggcaactggt attaatgctt ggttgcagga gttgccacag 83760
attgaacttg gtcgtgacca gaactctaaa acttggattt ttggttgtga gaagctagac 83820
tatacttcta caattccagc gtcaagtgtt taataaatcc tataaggggg agactttaaa 83880
aggtcttccc cttttttgtt tcttttaaaa gtattaagga atcacaatga aaacagaatt 83940
tagagtaatt aatggtaaga aagtaaatat cgttctgctt ggggcaagag atggtattaa 84000
gatgtctatg aagttgggta aaattgttgt tccaaccttt gcacagatgc tatcaagtct 84060
gactgacaag gataagaagg aagctccaat ggttccattt aaagaacttg ttgaagcttg 84120
ttttgacaga attgaagaga ttaaccttga agaaatggca actctgttat ttcaaggggc 84180
aactgttgat gacttcccac ttaatattga tacgtacttc caagcaaact atggtgagtt 84240
cattgattac ttagcatttg cgctggaggc aaacttcgga agttttttcg aagcaagcat 84300
tttcaaaagc ctaacttctc agtaaacatg ggcaacactc tacagacacc actaactgat 84360
gctgctgtag aggcaaccta tgaagaagca gacgagatga aatttgtgct tgctatttat 84420
ggtatggaag ggtgtaaaga aacacttgac caactctttg ctatgacatt ctctgattta 84480
ttatcattga gacaatttct tgaaattcag aggtcgtata aagaggaaat tgcttacaac 84540
gaacttagaa gagcaggaaa aatgtaatgg cacaatatac agttgatagc ttcattgtgg 84600
aacttggttt cagtgaaaag gttattaagg gcttgcaaag agttgagaag atgtctatgc 84660
aagctgctca acgtattgag agaaatatta ataaagcctt tgatgtgaaa cctaataaaa 84720
gctctcagga agcacttaat agaattgtaa aaaatgctca gtctgcttcg ggcagaatca 84780
ataaagcact caacagttcc ttgaaccttg attcacaagg tgttaaatct cttaagaaac 84840
ttgaaactca agcaaaaaag actgcaaaag gtattaacaa gtccttaaga gatgctatga 84900
atgttgacgg taaaatcact attaaaacag gcagagggag aggtggacag agcatccccc 84960
ctgttggtgg tggagcaccc agaggtcaga gagtagatgt tgctcagaga caaatggaaa 85020
ggatgtttaa taacaacttc tattcagggt taacccgtag actggaaaca attggtggtc 85080
aaggtaacca gatggcagcc tctttcagag ggagcttgca aaatatctac aacagataca 85140
aagggactgg taaggttggt gagtatgaga tggaagttaa aaagctcatc gacgtaacca 85200
aacgttgggt tattgcagaa aatgctagat taaaatcagt taaagaatca gcttggctac 85260
aggatagagc taacgcatca ttacgccaat tagttggtgg atttgtttca gcttacgctt 85320
tactggaact atctcaaaag actattgaag ctggtgtaaa aagacaatct gcacagttag 85380
cctctacagc tatctttgga gcagatacac agcaagctag aatgttcgct gcatcattcg 85440
cacaccagat tggtcagaac tacacagaca ctatgaagca gtactcaaac tttgctgctg 85500
gtgctcaacc aacacttggt tttcaaggta ctcaagagtt ctataagaat gctgcaatgt 85560
ttgcccgtat caggggtgct actgatgaag acttgaaagg tatcatggtt gcattccagc 85620
agatggcatc aaaaggtaaa gttcaggctg aagaactccg tggacagtta ggtgaccgtt 85680
tagcaggtgc tgtacagcta ttcgctgatg ccattggtaa gaccccacaa gaacttgata 85740
agatgatgca agatggtaaa ctgcttgctc aagacgttct tcctaaagta tctgaaagaa 85800
tggctgaact tgtcaagcaa gcaggtggta tgaacgctgt atctaagcag accgctacat 85860
caatgggtca agctaaggca atgtgggata acacacttgt agcactgttt aacaactcta 85920
gtgagggtat ttcacagtta tctaactctg ttgcaatgtt cttacaaggt tctatgggta 85980
gcacacaagc tttgggtctt gtgattggga gtctcttaaa aggtgctagt aatctacttg 86040
acttcgttac agacttcatg tacagaacat ctgcactata ctactatgca agagcttggt 86100
ataaagacct tgacaacagc cagcaaaagc ttatcaaaag tgctggtgaa tttctaggaa 86160
cagttgtcac aattggcggc gcagttgctg tagtatcaaa agcagtcaag ctcctaagtg 86220
gtttggttgg tgggggtatc tttggtaaaa tcttacaaag actcggtgtt agtgcagcag 86280
gtacagcagc agccggagaa gcagccgcag cagcaggtgg agttacagca acgagaatgg 86340
cacttggcac tgttggttcg gcattaatgc taagaggttc tacagaccca aatgctgcta 86400
aaaactacag tgaagtcaca ttaccaaaac catttgaaaa tgctgttgct aatattacaa 86460
acccaaaaag accaatgttc tttgatgaga atggtcaact tcagtttgca cagtacactc 86520
aagacgttga aggtaacagg aagttaattg ataatggtct atctaattgg gagattatca 86580
tggagaagct atcaacatct attgataatt ttgccaataa gtttaaccag acaccaatga 86640
tgatgacacc ttctggttta cctatgcaga ctaaacaaac cctgaatgtt actttcaatc 86700
ttgatggtaa acaaattgct actaagatgg tggatattac tgacaagaat caagaagaca 86760
ttcttctaag ttcaagctat ccagaggaag aataatgtta tgggattcta atatgcaaat 86820
caaatatagt ggcaaagatg gcatctattt ccacttaaga gataatgtag atgccttctt 86880
aaccttatca gcaactgaaa atatggaatt tgacagccct atgcaggtaa ctacacagaa 86940
catgcaatca gggcaaaccg tcacagataa tgtgcaaaga gcacccagaa caatcactat 87000
tagtggtgtc gttgtagttg gctatgaagg tagcttatta ttaactcgtc agggacaatt 87060
agtagaaaac ttcattgaca ctcttgaaaa ctggcgtgac cagaaacaaa ttatttcggt 87120
catctgtaaa gacggtatca agattgacga ttccattatt acaagcttta aagcctctaa 87180
agatgttgga atttctaatg gactaagaat ccagctaact tttcaggaaa ttaacttcaa 87240
agctattgta gggcaaactg atgtttcagc agctactgga aagaccgcta ccacaaatga 87300
tggtggcgca acttctaaga aaaatacggg taacactaca acaagtttag gtaatggtaa 87360
actgaattgc cagttactgt ttgacctaga tgctaatggt gtaagggaac tcactagtgc 87420
tgaagacaat gcgcttggga aatgctcaat gtctgcgaag acaagaaaag gtgttaccac 87480
attcagtgat gaggctgaaa aaaatgccgg agcagcatta aataggactg ctggaactgg 87540
taaggcatta caaaagcact cagtgaatcc gaataagaag gggacttatt a 87591
Claims (10)
1.一株宽裂解谱沙门氏菌噬菌体RDP-NSA-19050,其特征在于,透过电镜观察,该噬菌体头部为二十面体,长径约60nm,横径约70nm,尾长约100nm,直径约10nm,为有尾病毒目肌尾病毒科;基因全长87591bp,不具有毒力基因和溶原基因,保藏编号CGMCC No.21415。
2.如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050,其特征在于,该噬菌体在pH在5-9范围内作用1小时,其效价在1×1010pfu/mL之上,适宜生长在中性环境中;在pH=12以及pH=2时,仍有噬菌体存活,该噬菌体的酸碱耐受范围宽。
3.如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050,其特征在于,该噬菌体在35℃-60℃内活性保持稳定,效价在1010.5pfu/mL以上,在90℃高温条件下作用30min,依然具有生物活性,对温度有较好的耐受性。
4.如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050,其特征在于,该噬菌体RDP-NSA-19050高效感染变形杆菌,其最佳感染复数为1:10。
5.如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050在制备预防和或治疗沙门氏菌引起的疾病的药物中的用途。
6.如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050在制备预防和或治疗沙门氏菌引起的疾病的饲料中的用途。
7.一种用于防治沙门氏菌的杀菌组合物,其特征在于,包括有效量的如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050。
8.如权利要求7所述的用于防治沙门氏菌的杀菌组合物,其特征在于,还包括用于抗沙门氏菌的抗生素。
9.一种预防和或治疗沙门氏菌引起的疾病的添加剂,其有效成分包括权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050。
10.一种清洁剂或消毒剂,其特征在于,包括如权利要求1所述的沙门氏菌噬菌体RDP-NSA-19050。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110053070.3A CN112831474B (zh) | 2021-01-15 | 2021-01-15 | 一株宽裂解谱沙门氏菌噬菌体rdp-nsa-19050及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110053070.3A CN112831474B (zh) | 2021-01-15 | 2021-01-15 | 一株宽裂解谱沙门氏菌噬菌体rdp-nsa-19050及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112831474A true CN112831474A (zh) | 2021-05-25 |
CN112831474B CN112831474B (zh) | 2022-04-22 |
Family
ID=75928220
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110053070.3A Active CN112831474B (zh) | 2021-01-15 | 2021-01-15 | 一株宽裂解谱沙门氏菌噬菌体rdp-nsa-19050及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112831474B (zh) |
Cited By (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113416712A (zh) * | 2021-06-30 | 2021-09-21 | 华中农业大学 | 一株宽裂解谱沙门氏菌噬菌体及其应用 |
CN113430176A (zh) * | 2021-08-05 | 2021-09-24 | 瑞科盟(青岛)生物工程有限公司 | 一株稳定高效的烈性沙门氏菌噬菌体rdp-sa-21004及其应用 |
CN113583971A (zh) * | 2021-08-05 | 2021-11-02 | 瑞科盟(青岛)生物工程有限公司 | 一株可同时裂解大肠杆菌的沙门氏菌噬菌体及其应用 |
CN113913391A (zh) * | 2021-08-05 | 2022-01-11 | 瑞科盟(青岛)生物工程有限公司 | 一株可跨种裂解的大肠埃希氏菌噬菌体rdp-ec-20128及其应用 |
CN114107222A (zh) * | 2021-11-23 | 2022-03-01 | 华中农业大学 | 一株广谱耐高温沙门菌烈性噬菌体及其应用 |
CN114292836A (zh) * | 2021-11-05 | 2022-04-08 | 广东医科大学 | 一种内切沙门氏菌噬菌体的裂解酶、其编码基因及其制备方法和应用 |
CN115896041A (zh) * | 2022-10-24 | 2023-04-04 | 华中农业大学 | 一株兼性裂解沙门氏菌和大肠杆菌的广谱噬菌体和应用 |
CN115927206A (zh) * | 2022-07-28 | 2023-04-07 | 青岛润达生物科技有限公司 | 一株跨种属裂解型克罗诺杆菌噬菌体及其用途 |
CN117701512A (zh) * | 2024-02-06 | 2024-03-15 | 江苏大方生物工程有限公司 | 一种沙门菌噬菌体及其应用 |
CN118165942A (zh) * | 2024-03-22 | 2024-06-11 | 青岛润达生物科技有限公司 | 一株沙门氏菌噬菌体rdp-sa-22022及其连续化生产工艺和应用 |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013024304A1 (en) * | 2011-08-17 | 2013-02-21 | The University Of Nottingham | Bacteriophages |
EP2655603A2 (en) * | 2010-12-21 | 2013-10-30 | CJ CheilJedang Corporation | Novel bacteriophage and antibacterial composition comprising the same |
US20140017205A1 (en) * | 2009-09-03 | 2014-01-16 | Cj Cheiljedang Corporation | Bacteriophage and antibacterial composition comprising the same |
CN108359644A (zh) * | 2018-02-07 | 2018-08-03 | 青岛诺安百特生物技术有限公司 | 一种宽谱沙门氏菌噬菌体及其应用 |
CN111100844A (zh) * | 2019-08-09 | 2020-05-05 | 青岛润达生物科技有限公司 | 一株高裂解率沙门氏菌噬菌体rdp-sa-17118的分离及应用 |
CN112063594A (zh) * | 2020-09-24 | 2020-12-11 | 瑞科盟(青岛)生物工程有限公司 | 一株耐高温沙门氏菌噬菌体rdp-sa-18056及其微胶囊的制备工艺 |
-
2021
- 2021-01-15 CN CN202110053070.3A patent/CN112831474B/zh active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20140017205A1 (en) * | 2009-09-03 | 2014-01-16 | Cj Cheiljedang Corporation | Bacteriophage and antibacterial composition comprising the same |
EP2655603A2 (en) * | 2010-12-21 | 2013-10-30 | CJ CheilJedang Corporation | Novel bacteriophage and antibacterial composition comprising the same |
WO2013024304A1 (en) * | 2011-08-17 | 2013-02-21 | The University Of Nottingham | Bacteriophages |
CN108359644A (zh) * | 2018-02-07 | 2018-08-03 | 青岛诺安百特生物技术有限公司 | 一种宽谱沙门氏菌噬菌体及其应用 |
CN111100844A (zh) * | 2019-08-09 | 2020-05-05 | 青岛润达生物科技有限公司 | 一株高裂解率沙门氏菌噬菌体rdp-sa-17118的分离及应用 |
CN112063594A (zh) * | 2020-09-24 | 2020-12-11 | 瑞科盟(青岛)生物工程有限公司 | 一株耐高温沙门氏菌噬菌体rdp-sa-18056及其微胶囊的制备工艺 |
Non-Patent Citations (1)
Title |
---|
包红朵等: "沙门氏菌裂解性噬菌体的分离鉴定及其生物学特性", 《食品科学》 * |
Cited By (16)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113416712B (zh) * | 2021-06-30 | 2022-04-26 | 华中农业大学 | 一株宽裂解谱沙门氏菌噬菌体及其应用 |
CN113416712A (zh) * | 2021-06-30 | 2021-09-21 | 华中农业大学 | 一株宽裂解谱沙门氏菌噬菌体及其应用 |
CN113430176A (zh) * | 2021-08-05 | 2021-09-24 | 瑞科盟(青岛)生物工程有限公司 | 一株稳定高效的烈性沙门氏菌噬菌体rdp-sa-21004及其应用 |
CN113583971A (zh) * | 2021-08-05 | 2021-11-02 | 瑞科盟(青岛)生物工程有限公司 | 一株可同时裂解大肠杆菌的沙门氏菌噬菌体及其应用 |
CN113913391A (zh) * | 2021-08-05 | 2022-01-11 | 瑞科盟(青岛)生物工程有限公司 | 一株可跨种裂解的大肠埃希氏菌噬菌体rdp-ec-20128及其应用 |
CN113583971B (zh) * | 2021-08-05 | 2022-06-28 | 瑞科盟(青岛)生物工程有限公司 | 一株可同时裂解大肠杆菌的沙门氏菌噬菌体及其应用 |
CN114292836A (zh) * | 2021-11-05 | 2022-04-08 | 广东医科大学 | 一种内切沙门氏菌噬菌体的裂解酶、其编码基因及其制备方法和应用 |
CN114107222A (zh) * | 2021-11-23 | 2022-03-01 | 华中农业大学 | 一株广谱耐高温沙门菌烈性噬菌体及其应用 |
CN114107222B (zh) * | 2021-11-23 | 2023-07-11 | 华中农业大学 | 一株广谱耐高温沙门菌烈性噬菌体及其应用 |
CN115927206B (zh) * | 2022-07-28 | 2024-09-24 | 青岛润达生物科技有限公司 | 一株跨种属裂解型克罗诺杆菌噬菌体及其用途 |
CN115927206A (zh) * | 2022-07-28 | 2023-04-07 | 青岛润达生物科技有限公司 | 一株跨种属裂解型克罗诺杆菌噬菌体及其用途 |
CN115896041A (zh) * | 2022-10-24 | 2023-04-04 | 华中农业大学 | 一株兼性裂解沙门氏菌和大肠杆菌的广谱噬菌体和应用 |
CN115896041B (zh) * | 2022-10-24 | 2024-05-03 | 华中农业大学 | 一株兼性裂解沙门氏菌和大肠杆菌的广谱噬菌体和应用 |
CN117701512B (zh) * | 2024-02-06 | 2024-06-11 | 江苏大方生物工程有限公司 | 一种沙门菌噬菌体及其应用 |
CN117701512A (zh) * | 2024-02-06 | 2024-03-15 | 江苏大方生物工程有限公司 | 一种沙门菌噬菌体及其应用 |
CN118165942A (zh) * | 2024-03-22 | 2024-06-11 | 青岛润达生物科技有限公司 | 一株沙门氏菌噬菌体rdp-sa-22022及其连续化生产工艺和应用 |
Also Published As
Publication number | Publication date |
---|---|
CN112831474B (zh) | 2022-04-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN112831474B (zh) | 一株宽裂解谱沙门氏菌噬菌体rdp-nsa-19050及其应用 | |
CN113583972B (zh) | 一株可降低抗生素耐药性的大肠杆菌噬菌体及其应用 | |
CN112063594B (zh) | 一株耐高温沙门氏菌噬菌体rdp-sa-18056及其微胶囊的制备工艺 | |
CN112195158B (zh) | 一株裂解性副溶血性弧菌噬菌体rdp-vp-19003及其应用 | |
CN112760296B (zh) | 一株副溶血弧菌噬菌体rdp-vp-19010及其应用 | |
CN113430174B (zh) | 一株高裂解性沙门氏菌噬菌体rdp-sa-19009及其应用 | |
CN112662635B (zh) | 一株耐高温型大肠杆菌噬菌体rdp-ec-20031及其应用 | |
CN107250352A (zh) | 新型肠致病性大肠杆菌噬菌体Esc‑CHP‑2及其用于抑制肠致病性大肠杆菌增殖的用途 | |
KR101609111B1 (ko) | 신규한 에드와드시엘라 타르다 박테리오파지 EdW-TAP-1 및 이의 에드와드시엘라 타르다 균 증식 억제 용도 | |
CN113430173B (zh) | 一株宽裂解谱、耐高温沙门氏菌噬菌体及其应用 | |
CN112771156A (zh) | 大肠杆菌噬菌体esc-cop-14及其在抑制致病性大肠杆菌增殖方面的用途 | |
CN112708600A (zh) | 一株裂解性肺炎克雷伯菌噬菌体rdp-kp-20005及其应用 | |
KR101915486B1 (ko) | 신규한 비브리오 파라헤몰리티쿠스 박테리오파지 Vib-PAP-7 및 이의 비브리오 파라헤몰리티쿠스 균 증식 억제 용도 | |
CN108179136B (zh) | 大肠杆菌噬菌体Esc-COP-9及其致病性大肠杆菌增殖抑制用途 | |
CN108179135B (zh) | 大肠杆菌噬菌体Esc-COP-7及其致病性大肠杆菌增殖抑制用途 | |
CN113166730A (zh) | 新型猪链球菌噬菌体str-sup-1及其在抑制猪链球菌细菌增殖方面的用途 | |
CN113528463A (zh) | 一株裂解性、高效价副溶血弧菌噬菌体rdp-vp-21010及其应用 | |
KR102066920B1 (ko) | 신규한 크로노박터 사카자키 특이 박테리오파지 cs3 및 이를 포함하는 항균 조성물 | |
KR101863593B1 (ko) | 신규한 살모넬라균 특이 박테리오파지 sp1 및 이를 포함하는 항균 조성물 | |
CN113322242B (zh) | 一株多重耐药性大肠杆菌噬菌体及其应用 | |
CN112522214B (zh) | 一株高裂解性铜绿假单胞菌噬菌体rdp-pa-20001及其应用 | |
KR102193495B1 (ko) | 신규한 아시네토박터 바우마니 균 특이 박테리오파지 ab63 및 이를 포함하는 항균 조성물 | |
CN115942947A (zh) | 用于抑制致病性大肠杆菌增殖的组合物和方法 | |
KR20180102040A (ko) | 신규한 비브리오 파라헤몰리티쿠스 박테리오파지 Vib-PAP-4 및 이의 비브리오 파라헤몰리티쿠스 균 증식 억제 용도 | |
CN112646785B (zh) | 一株耐高温烈性变形杆菌噬菌体rdp-sa-20018及其应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |