CN114807201B - 一种纤维寡糖转运蛋白及其制备方法和应用 - Google Patents
一种纤维寡糖转运蛋白及其制备方法和应用 Download PDFInfo
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
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- C12N15/70—Vectors or expression systems specially adapted for E. coli
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- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
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- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/102—Mutagenizing nucleic acids
- C12N15/1024—In vivo mutagenesis using high mutation rate "mutator" host strains by inserting genetic material, e.g. encoding an error prone polymerase, disrupting a gene for mismatch repair
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- C—CHEMISTRY; METALLURGY
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Abstract
本发明提供一种纤维寡糖转运蛋白及其制备方法和应用，该纤维寡糖转运蛋白工程菌株的制备方法包括：步骤1，以野生型SP‑LacY作为初始噬菌体，以包含AP1质粒、AP2质粒、MP1质粒的可视化噬菌体辅助持续定向进化系统用工程菌作为宿主，以含纤维寡糖的LB或M9软琼脂为培养基培养噬菌体斑；步骤2，将获得的噬菌体斑继续进行多轮进化培养，对每轮进化获得的噬菌体斑测序以验证突变体累积趋势，直至没有新的突变体形成即可停止进化，最终获得具有不同突变位点的蛋白；步骤3，将步骤2获得的蛋白的不同突变位点进行组合来构建不同的LacY突变体表达载体，再将LacY突变体表达载体转化宿主菌，即得。本发明的纤维寡糖转运蛋白能更好的发酵利用纤维素及纤维寡糖。
Description
技术领域
本发明属于基因工程技术领域，具体涉及一种纤维寡糖转运蛋白及其制备方法和应用。
背景技术
纤维质是世界上蕴藏量最丰富的天然高分子化合物，绝大多数由绿色植物通过光合作用合成。当前利用纤维质生产工业副产品，如酒精，首先需将纤维质进行理化处理或者用纤维素酶等预处理将纤维质降解成葡萄糖，再供微生物利用生产工业副产品；事实上经过预处理的纤维质降解产物中还含有大量的纤维二糖、纤维三糖等纤维寡糖，但是微生物中由于缺乏特异性的纤维寡糖转运蛋白，微生物无法高效利用纤维寡糖，导致资源浪费，纤维质利用率低。
LacY是乳糖或半乳糖转运蛋白，其对纤维寡糖的转运效率非常低，且当前在细菌中并未发现特异性转运纤维寡糖的转运蛋白，因此在纤维质利用领域，尤其是利用细菌发酵纤维质获取工业副产品方面受到制约，如能开发出一种高效转运纤维寡糖进胞内供细菌利用的转运蛋白即能够解决上述问题。
发明内容
针对现有技术存在的问题和缺陷，本发明提供一种纤维寡糖转运蛋白及其制备方法和应用，该纤维寡糖转运蛋白具有高效纤维寡糖转运效率。本发明的技术方案为：
第一个方面，本发明提供一种纤维寡糖转运蛋白工程菌株的制备方法，包括以下步骤：
步骤1，以野生型SP-LacY作为初始噬菌体，以包含AP1质粒、AP2质粒、MP1质粒的可视化噬菌体辅助持续定向进化系统用工程菌作为宿主，以含纤维寡糖的LB或M9软琼脂为培养基培养噬菌体斑；
步骤2，将获得的噬菌体斑继续进行多轮进化培养，多轮进化培养过程逐步调低纤维寡糖在培养基中的浓度，对每轮进化获得的噬菌体斑测序以验证突变体累积趋势，直至没有新的突变体形成即可停止进化，最终获得具有不同突变位点的蛋白；
所述AP1质粒、所述AP2质粒、所述MP1质粒的核酸序列分别如SEQ ID NO.2、SEQ IDNO.3、SEQ ID NO.4所示；
步骤3，将步骤2获得的蛋白的不同突变位点进行组合来构建不同的LacY突变体表达载体，再将LacY突变体表达载体转化宿主菌，即得纤维寡糖转运蛋白工程菌株。
进一步地，所述步骤1的具体操作过程为：所述初始噬菌体的滴度为1×109pfu/mL，将宿主点在培养基正中央，在离宿主位点1～1.5cm处分别点三个噬菌体液滴，每个液滴的体积为1-2uL。
进一步地，所述野生型SP-LacY的核酸序列如SEQ ID NO.5所示。
进一步地，所述可视化噬菌体辅助持续定向进化系统用工程菌为AP1质粒、AP2质粒、MP1质粒共转化携带F因子的大肠杆菌得到的工程菌株。
优选地，所述携带F因子的大肠杆菌为大肠杆菌FM15，其构建方法包括：将MG1655大肠杆菌菌株与K12 ER2738(NEB公司)杂交，赋予MG1755菌株F因子；用λRed敲除技术敲除LacZ基因α片段，所得菌株即为FM15，其基因型为F’proA+B+lacIqΔ(lacZ)M15 zzf::Tn10(TetR)/λ–ilvG–rfb-50rph-1attB::KanRΔ(lacZ)M15。
可选地，AP1质粒、AP2质粒、MP1质粒可通过重新构建放在一个质粒或两个质粒上以精简系统。
进一步地，所述AP1质粒携带gIII基因并由pPSP启动子启动，pPSP启动子与核糖体结合位点RBS之间有功能蛋白CelR的识别位点CelRS，CelRS的核酸序列如SEQ ID NO:6所示。
进一步地，所述AP2质粒携带功能蛋白CelR的蛋白基因和pPSP启动子的调控蛋白基因。
进一步地，所述诱变质粒MP1由pPSP启动子启动。
进一步地，所述步骤1中纤维寡糖在培养基中的初始浓度为1％(g/mL)。
进一步地，所述步骤3中宿主菌为大肠杆菌，优选为S1030大肠杆菌。
第二个方面，本发明提供一种纤维寡糖转运蛋白，是由上述制备方法获得。
进一步地，所述纤维寡糖包括纤维二糖、纤维三糖、纤维四糖。
第三个方面，本发明提供上述纤维寡糖转运蛋白或者上述制备方法获得的纤维寡糖转运蛋白工程菌株在纤维素发酵中的应用。
本发明的有益效果为：本发明采用包含AP1质粒、AP2质粒、MP1质粒的大肠杆菌作为宿主，进行可视化噬菌体辅助持续定向进化膜蛋白LacY，对携带野生型LacY基因的噬菌体SP-LacY进行多轮进化突变，获得的纤维寡糖转运蛋白能大大提高纤维寡糖转运效率，从而更好的发酵利用纤维素及纤维寡糖。
附图说明
图1为本发明实施例1中培养噬菌体斑的点板示意图。
图2为本发明实施例1中LacY突变体表达载体P1图谱(携带野生型LacY)。
图3为本发明实施例2中携带CDP基因的表达载体P2图谱。
图4为本发明实施例3中不同LacY突变体的大肠杆菌消耗纤维二糖的耗糖曲线。
图5为本发明实施例4中不同LacY突变体的大肠杆菌消耗纤维三糖的耗糖曲线。
图6为本发明实施例6中不同LacY突变体的大肠杆菌消耗纤维四糖的耗糖曲线。
具体实施方式
在本发明的描述中，需要说明的是，实施例中未注明具体条件者，按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者，均为可以通过市售购买获得的常规产品。
下面结合附图和具体的实施例对本发明做进一步详细说明，所述是对本发明的解释而不是限定。
实施例1
本实施例提供一种纤维二糖转运蛋白工程菌株的制备方法，具体如下：
步骤1，可视化噬菌体辅助持续定向进化系统的搭建：
该系统包含AP1、AP2、MP1三个质粒，也可以将三个质粒上的功能元件重排放在两个质粒或者一个质粒上；AP1质粒(SEQ ID NO:2)携带gIII基因并由pPSP启动子启动，pPSP启动子与核糖体结合位点RBS之间有功能蛋白CelR的识别位点CelRS(SEQ ID NO.6)，在胞内没有纤维二糖的情况下，gIII表达处于抑制状态。AP2质粒(SEQ ID NO:3)携带CelR蛋白基因和启动子pPSP的调控蛋白基因。诱变质粒MP1(SEQ ID NO：4)由pPSP启动子启动。宿主菌FM15由本实验自己构建，其构建方法包括：按照分子生物学常规操作将MG1655大肠杆菌菌株与K12 ER2738(NEB公司)杂交，赋予MG1755菌株F因子；用常规λRed敲除技术敲除LacZ基因α片段，所得菌株即为FM15，其基因型为F’proA+B+lacIqΔ(lacZ)M15 zzf::Tn10(TetR)/λ–ilvG–rfb-50rph-1attB::KanRΔ(lacZ)M15。本发明中的宿主菌并不限于FM15，但必须是携带F因子的大肠杆菌。AP1质粒、AP2质粒、MP1质粒共转化F15菌株感受态，得到进化用的工程菌株FM15-AP1/AP2/MP1。
步骤2，培养噬菌体斑：
初始噬菌体为野生型SP-lacY(SEQ ID NO.5)，滴度1×109pfu/mL，宿主为FM15-AP1/AP2/MP1(OD600＝0.4)，宿主菌1-2uL点在LB软琼脂(含1％的纤维二糖)正中央，在离宿主位点大概1cm处分别点三个噬菌体液滴(1-2uL)，隔天即可看见三个大的噬菌体扇形斑(图1)。
步骤3，噬菌体斑继续多轮进化培养：
将获得的噬菌体斑经过多轮进化培养，第二轮类似，只是将纤维二糖在培养基中的浓度降低为0.5％，噬菌体用第一轮进化后的噬菌体稀释100倍点板；之后的轮次以此类推，每轮进化培养中纤维二糖在培养基中的浓度降低为前一轮的1/2，或者根据斑大小调整浓度，直到突变体库不在发生变化为止。每轮取样测序检测LacY突变累计情况，共计进行了13轮进化培养。
结果显示针对纤维二糖的进化，先后出现L70H、G71V、A177V、G202V、N204S、V197A、A213P、F356S几个突变位点的累积(原LacY的氨基酸序列如SEQ ID NO.1所示)。
步骤4，工程菌株的构建：
将上述不同的突变位点组合构建不同的LacY突变体表达载体P1(图2，SEQ IDNO.7)，LacY的启动子为J23109，cepA的启动子为J23102，cepA是纤维二糖磷酸化酶，可以直接将LacY转运进胞内的纤维二糖降解为葡萄糖和1磷酸葡萄糖供细菌代谢利用；上述启动子不局限于此，可以用其他启动子代替。此外，载体的构建是分子生物学基本技能，本领域技术人员根据习惯或手册即可完成。
再将LacY突变体表达载体转化宿主菌，即得纤维二糖转运蛋白工程菌株。所用表达宿主菌为LacY基因缺陷型大肠杆菌S1030[David R Liu实验室提供，已有相关文献报道其遗传信息(Nat Chem Biol.2014March；10(3):216–222)]，可通过常规基因操作获取；宿主也可以由其他商业化的LacY基因缺陷型的大肠杆菌代替。表达载体电转或化转S1030感受态细胞，通过壮观霉素筛选阳性克隆，测序验证正确获得不同LacY突变体的工程菌株；大肠杆菌的感受态制备及转化方法是本领域的基本技能，本领域技术人员容易获得。
实施例2
本实施例提供一种纤维三糖转运蛋白工程菌株的制备方法，步骤1～3同实施例1，用与实施例1相同的可视化PACE系统进化LacY对纤维三糖转运效率获得突变位点A177V、G202V、V197A。在步骤4工程菌株的构建过程中，纤维三糖的代谢额外需要纤维寡糖磷酸化酶CDP将其降解为1磷酸葡萄糖和纤维二糖，纤维二糖可继续被cepA降解完全供细菌利用。携带CDP基因的表达载体P2(SEQ ID NO.8)，见图3，将P1和P2共转化宿主S1030即可得到纤维三糖代谢用的纤维三糖转运蛋白工程菌株。
实施例3
LacY突变体工程菌株纤维二糖代谢实验
将实施例1中构建好的部分工程菌株S1030-cepA-LacYWT(野生型)、S1030-cepA-LacYA177V、S1030-cepA-LacYL70H、S1030-cepA-LacYA177V/V197A、S1030-cepA-LacYA177V/G202V、S1030-cepA-LacYA177V/N204S在M9培养基中培养并以纤维二糖为唯一碳源；测定不同工程菌株纤维二糖的消耗曲线，以此判断不同LacY突变体转运纤维二糖的效率。此外，由于G71V、A213P、F356S这几个突变位点出现时间较短，故被淘汰掉，不予考察。
从图4显示的结果可以看出，不同工程菌株的耗糖速率是：A177V/V197A>A177V，A177V/G202V，A177V/N204S>L70H>LacYWT。因此，不同工程菌株转运纤维二糖的效率排序为：A177V/V197A>A177V，A177V/G202V，A177V/N204S>L70H>LacYWT。
实施例4
LacY突变体工程菌株纤维三糖代谢实验
将实施例2中构建好的部分工程菌株S1030-CDP-cepA-LacYW(野生型)、S1030-CDP-cepA-LacYA177V(图5中显示为LacY177)、S1030-CDP-cepA-LacYA177V/V197A(图5中显示为LacY177197)在M9培养基中培养并以纤维三糖为唯一碳源；测定不同工程菌株纤维三糖的消耗曲线，以此判断不同LacY突变体转运纤维三糖的效率。此外，由于G202V突变位点出现时间较短，故被淘汰掉，不予考察。
从图5显示的结果可以看出，不同工程菌株的耗糖速率是：A177V/V197A>A177V>LacYW。因此，不同工程菌株转运纤维三糖的效率排序为：A177V/V197A>A177V>LacYW。
实施例5
本实施例提供一种纤维四糖转运蛋白工程菌株的制备方法，步骤1～3同实施例1，用与实施例1相同的可视化PACE系统进化LacY对纤维四糖转运效率获得突变位点A177V、V197A。在步骤4工程菌株的构建过程中，纤维四糖的代谢额外需要纤维寡糖磷酸化酶CDP将其降解为1磷酸葡萄糖和纤维二糖，纤维二糖可继续被cepA降解完全供细菌利用。携带CDP基因的表达载体P2(SEQ ID NO.8)，见图3，将P1和P2共转化宿主S1030即可得到纤维四糖代谢用的工程菌株。
实施例6
LacY突变体工程菌株纤维四糖代谢实验：由于对纤维四糖进化获得的突变位点刚好属于纤维三糖进化的一部分，进而考查的是工程菌株S1030-CDP-cepA-LacYW(野生型)、S1030-CDP-cepA-LacYA177V/V197A在M9培养基中培养并以纤维四糖为唯一碳源的代谢结果，图见实施例6，LacYA177/197的耗糖速率要大于野生型LacYW。
综上，本发明获得的纤维寡糖转运蛋白工程菌株能大大提高纤维寡糖转运效率，从而更好的发酵利用纤维素及纤维寡糖。
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细，但并不能因此而理解为对本发明专利范围的限制。应当指出的是，对于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。因此，本发明专利的保护范围应以所附权利要求为准。
序列表
<110> 广州先进技术研究所
<120> 一种纤维寡糖转运蛋白及其制备方法和应用
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Leu Leu Ser Asp Lys Leu Gly Leu Arg Lys Tyr Leu Leu Trp Ile Ile
65 70 75 80
Thr Gly Met Leu Val Met Phe Ala Pro Phe Phe Ile Phe Ile Phe Gly
85 90 95
Pro Leu Leu Gln Tyr Asn Ile Leu Val Gly Ser Ile Val Gly Gly Ile
100 105 110
Tyr Leu Gly Phe Cys Phe Asn Ala Gly Ala Pro Ala Val Glu Ala Phe
115 120 125
Ile Glu Lys Val Ser Arg Arg Ser Asn Phe Glu Phe Gly Arg Ala Arg
130 135 140
Met Phe Gly Cys Val Gly Trp Ala Leu Cys Ala Ser Ile Val Gly Ile
145 150 155 160
Met Phe Thr Ile Asn Asn Gln Phe Val Phe Trp Leu Gly Ser Gly Cys
165 170 175
Ala Leu Ile Leu Ala Val Leu Leu Phe Phe Ala Lys Thr Asp Ala Pro
180 185 190
Ser Ser Ala Thr Val Ala Asn Ala Val Gly Ala Asn His Ser Ala Phe
195 200 205
Ser Leu Lys Leu Ala Leu Glu Leu Phe Arg Gln Pro Lys Leu Trp Phe
210 215 220
Leu Ser Leu Tyr Val Ile Gly Val Ser Cys Thr Tyr Asp Val Phe Asp
225 230 235 240
Gln Gln Phe Ala Asn Phe Phe Thr Ser Phe Phe Ala Thr Gly Glu Gln
245 250 255
Gly Thr Arg Val Phe Gly Tyr Val Thr Thr Met Gly Glu Leu Leu Asn
260 265 270
Ala Ser Ile Met Phe Phe Ala Pro Leu Ile Ile Asn Arg Ile Gly Gly
275 280 285
Lys Asn Ala Leu Leu Leu Ala Gly Thr Ile Met Ser Val Arg Ile Ile
290 295 300
Gly Ser Ser Phe Ala Thr Ser Ala Leu Glu Val Val Ile Leu Lys Thr
305 310 315 320
Leu His Met Phe Glu Val Pro Phe Leu Leu Val Gly Cys Phe Lys Tyr
325 330 335
Ile Thr Ser Gln Phe Glu Val Arg Phe Ser Ala Thr Ile Tyr Leu Val
340 345 350
Cys Phe Cys Phe Phe Lys Gln Leu Ala Met Ile Phe Met Ser Val Leu
355 360 365
Ala Gly Asn Met Tyr Glu Ser Ile Gly Phe Gln Gly Ala Tyr Leu Val
370 375 380
Leu Gly Leu Val Ala Leu Gly Phe Thr Leu Ile Ser Val Phe Thr Leu
385 390 395 400
Ser Gly Pro Gly Pro Leu Ser Leu Leu Arg Arg Gln Val Asn Glu Val
405 410 415
Ala
<210> 2
<211> 5684
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
gggtacccat gatgaaattc gccacttgtt agtgtaattc gctaactcat cctggcatgt 60
tgctgttgat tcttcaatca gatctttata aatcaaaaag ataaaaaatt ggcacgcaaa 120
ttgtattaac agttcagcag gacaatcctg aattctggga gcgctcccat cacacaggaa 180
acagctatga aaaaattatt attcgcaatt cctttagttg ttcctttcta ttctcactcc 240
gctgaaactg ttcatcacca tcaccatcac gctgaaactg ttgaaagttg tttagcaaaa 300
ccccatacag aaaattcatt tactaacgtc tggaaagacg acaaaacttt agatcgttac 360
gctaactatg agggctgtct gtggaatgct acaggcgttg tagtttgtac tggtgacgaa 420
actcagtgtt acggtacatg ggttcctatt gggcttgcta tccctgaaaa tgagggtggt 480
ggctctgagg gtggcggttc tgagggtggc ggttctgagg gtggcggtac taaacctcct 540
gagtacggtg atacacctat tccgggctat acttatatca accctctcga cggcacttat 600
ccgcctggta ctgagcaaaa ccccgctaat cctaatcctt ctcttgagga gtctcagcct 660
cttaatactt tcatgtttca gaataatagg ttccgaaata ggcagggggc attaactgtt 720
tatacgggca ctgttactca aggcactgac cccgttaaaa cttattacca gtacactcct 780
gtatcatcaa aagccatgta tgacgcttac tggaacggta aattcagaga ctgcgctttc 840
cattctggct ttaatgagga tccattcgtt tgtgaatatc aaggccaatc gtctgacctg 900
cctcaacctc ctgtcaatgc tggcggcggc tctggtggtg gttctggtgg cggctctgag 960
ggtggtggct ctgagggtgg cggttctgag ggtggcggct ctgagggagg cggttccggt 1020
ggtggctctg gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct 1080
atgaccgaaa atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct 1140
gtcgctactg attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct 1200
aatggtaatg gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt 1260
gacggtgata attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa 1320
tcggttgaat gtcgcccttt tgtctttggc gctggtaaac cttacgagtt cagtatcgac 1380
tgcgataaga tcaacctgtt ccgcggtgtc tttgcgtttc ttttatatgt tgccaccttt 1440
atgtatgtat tttctacgtt tgctaacata ctgcgtaata aggagtctta atgaaatttg 1500
gaaacttttt gcttacatac caacctcccc aattttccca aacagaggta atgaaacgtt 1560
tggttaaatt aggtcgcatc tctgatgagt gtggttttga taccgtatgg ttactggagc 1620
atcatttcac ggagtttggt ttgcttggta acccttatgt cgctgctgca tatttacttg 1680
gcgcgactaa aaaattgaat gtaggaactg ccgctattgt tcttcccaca gcccatccag 1740
tacgccaact tgaagatgtg aatttattgg atcaaatgtc aaaaggacga tttcggtttg 1800
gtatttgccg agggctttac aacaaggact ttcgcgtatt cggcacagat atgaataaca 1860
gtcgcgcctt agcggaatgc tggtacgggc tgataaagaa tggcatgaca gagggatata 1920
tggaagctga taatgaacat atcaagttcc ataaggtaaa agtaaacccc gcggcgtata 1980
gcagaggtgg cgcaccggtt tatgtggtgg ctgaatcagc ttcgacgact gagtgggctg 2040
ctcaatttgg cctaccgatg atattaagtt ggattataaa tactaacgaa aagaaagcac 2100
aacttgagct ttataatgaa gtggctcaag aatatgggca cgatattcat aatatcgacc 2160
attgcttatc atatataaca tctgtagatc atgactcaat taaagcgaaa gagatttgcc 2220
ggaaatttct ggggcattgg tatgattctt atgtgaatgc tacgactatt tttgatgatt 2280
cagaccaaac aagaggttat gatttcaata aagggcagtg gcgtgacttt gtattaaaag 2340
gacataaaga tactaatcgc cgtattgatt acagttacga aatcaatccc gtgggaacgc 2400
cgcaggaatg tattgacata attcaaaaag acattgatgc tacaggaata tcaaatattt 2460
gttgtggatt tgaagctaat ggaacagtag acgaaattat tgcttccatg aagctcttcc 2520
agtctgatgt catgccattt cttaaagaaa aacaacgttc gctattatat tatggcggtg 2580
gcggtagcgg cggtggcggt agcggcggtg gcggtagcgg cggtggcggt agcaaatttg 2640
gattgttctt ccttaacttc atcaattcaa caactgttca agaacagagt atagttcgca 2700
tgcaggaaat aacggagtat gttgataagt tgaattttga acagatttta gtgtatgaaa 2760
atcatttttc agataatggt gttgtcggcg ctcctctgac tgtttctggt tttctgctcg 2820
gtttaacaga gaaaattaaa attggttcat taaatcacat cattacaact catcatcctg 2880
tccgcatagc ggaggaagct tgcttattgg atcagttaag tgaagggaga tttattttag 2940
ggtttagtga ttgcgaaaaa aaagatgaaa tgcatttttt taatcgcccg gttgaatatc 3000
aacagcaact atttgaagag tgttatgaaa tcattaacga tgctttaaca acaggctatt 3060
gtaatccaga taacgatttt tatagcttcc ctaaaatatc tgtaaatccc catgcttata 3120
cgccaggcgg acctcggaaa tatgtaacag caaccagtca tcatattgtt gagtgggcgg 3180
ccaaaaaagg tattcctctc atctttaagt gggatgattc taatgatgtt agatatgaat 3240
atgctgaaag atataaagcc gttgcggata aatatgacgt tgacctatca gagatagacc 3300
atcagttaat gatattagtt aactataacg aagatagtaa taaagctaaa caagagacgc 3360
gtgcatttat tagtgattat gttcttgaaa tgcaccctaa tgaaaatttc gaaaataaac 3420
ttgaagaaat aattgcagaa aacgctgtcg gaaattatac ggagtgtata actgcggcta 3480
agttggcaat tgaaaagtgt ggtgcgaaaa gtgtattgct gtactttgaa ccaatgaatg 3540
atttgatgag ccaaaaaaat gtaatcaata ttgttgatga taatattaag aagtaccaca 3600
cggaatatac ctaaacttaa ttaacggcac tcctcagcaa atataatgac cctcttgata 3660
acccaagagg gcatttttta atgcccatgg aagggcctcg tgatacgcct atttttatag 3720
gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg 3780
cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 3840
caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 3900
ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 3960
gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 4020
gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 4080
atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 4140
caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 4200
gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 4260
accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 4320
ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 4380
gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca 4440
acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta 4500
atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct 4560
ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca 4620
gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag 4680
gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat 4740
tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 4800
taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 4860
cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 4920
gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 4980
gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 5040
agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 5100
aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 5160
agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 5220
cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 5280
accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5340
aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5400
ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5460
cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 5520
gccgctaggt ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac 5580
agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctctaga 5640
ttaattaatt aagcggccgc atcgatcggg ccctgaggcc tgca 5684
<210> 3
<211> 5867
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
aaagagttgc tgcaactcag tttgcaacaa gggaaatata accagaagcg cgcggctgaa 60
ttactggggt taacttatca tcagttccgc gcgttgttga aaaagcacca gatttagact 120
agtaaaacga aaggcccagt ctttcgactg agcctttcgt tttatttggg tacctttaca 180
gctagctcag tcctagggac tgtgctagcg aattctagag aaagaggaga aactcgagat 240
ggaacgtcgc cgtcgcccga ccctggaaat ggttgcagcc ctggccggtg tctgtcgtgg 300
tacggtgagc cgcgttatta acggtagcga tcaggtctct ccggcgaccc gtgaagccgt 360
gaaacgcgca atcaaagaac tgggctatgt gccgaatcgt gcagctcgta ccctggtgac 420
ccgtcgtacc gatacggttg cactggtggt ttctgaaaac aatcagaaac tgtttgctga 480
accgttctac gcgggtattg tgctgggtgt tggtgtcgca ctgagcgaac gtggctttca 540
attcgttctg gcaaccggcc gttctggtat cgaacatgaa cgcctgggcg gttatctggc 600
aggccagcat gtcgatggtg tgctgctgct gtcactgcac cgcgatgacc cgctgccgca 660
aatgctggac gaagcgggcg ttccgtatgt ctatggcggt cgtccgctgg gtgtgccgga 720
agaacaggtg tcgtacgttg atattgacaa catcggtggt ggccgtcagg caacccaacg 780
tctgattgaa acgggtcacc gtcgtattgc aaccatcgca ggtccgcagg atatggtcgc 840
tggcgtggaa cgtctgcaag gttatcgcga agccctgctg gcggccggta tggaatacga 900
cgaaaccctg gttagttatg gcgattttac gtacgactcc ggtgtcgcag ctatgcgtga 960
actgctggat cgtgcgccgg atgttgacgc agtcttcgca gccagtgacc tgatgggcct 1020
ggcagctctg cgtgttctgc gtgcttccgg tcgtcgcgtc ccggaagatg tggcagtcgt 1080
gggttatgat gactcaaccg tggcagaaca tgctgaaccg ccgatgacct cggttaatca 1140
gccgacggaa ctgatgggtc gtgaaatggc gcgcctgctg gtggatcgta tcaccggtga 1200
aaccacggaa ccggtgcgcc tggttatgga aacgcacctg atggttcgtg aatcaggcta 1260
actgcaggtc cctaagtctc ctcagcaaaa cgaaaggccc agtctttcga ctgagccttt 1320
cgttttattt gaccggatgt cctcttgttc atcatcagta acccgtatcg tgagcatcct 1380
ctctcgtttc atcggtatca ttaccttttt ccataggctc cgcccccctg acaagcatca 1440
cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc 1500
gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt 1560
cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg 1620
cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct 1680
tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag 1740
cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa 1800
actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag 1860
ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag 1920
caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattac ggcctatttg 1980
gcctattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 2040
tgcttcaata atattgaaaa aggaagagta tgagggaagc ggtgatcgcc gaagtatcga 2100
ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg 2160
tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc 2220
tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt 2280
tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg 2340
ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag 2400
aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc 2460
tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg 2520
aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa 2580
cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt 2640
cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact 2700
gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt 2760
atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc 2820
actacgtgaa aggcgagatc accaaggtag tcggcaaata aacgccatgg caaataaaac 2880
gaaaggctca gtcgaaagac tgggcctttc gttttttaac ctttgacctt cggcattgcg 2940
atgtctttca ggccaccggc gttctcaacg tgggtatatc ccatctcgct aaggatctct 3000
tttgcttgcc ctgactggcg tccggcattg caatacactt tcacggtgtc gtttttatcc 3060
ggaacggcgg tggcaatgcg ctctttcact tccttcaggg gaatattgat ggccccctga 3120
acgtgctctt gctgatactg ctctggaaca cgaacatcga tccagtgttc agcggcgaaa 3180
acgggcagtg aaaacaccag tgccagagca agtaagcctt ttttaaacat gtaagcgtcc 3240
ttctatggat gaaaattgcc atccattata cgcctgagat ttaacaattt ttgcgaaact 3300
cccctcacct tttataacgc tgtgccagtt tattagcagc ccgacttaac agcggttcca 3360
gcgccacagc cagcaacatt ttcagcggtc ggcgagcaac tgattttatc gcccagcccg 3420
ccacacccgc cgggccatag cgcagtgcgg taagaagtac cagcttgcct gctaatttga 3480
aaccaggctt taccttttgc ccggcctgtt gccagcgagt attcataact ttcctcacag 3540
ttgacggaaa cggctacgta acgtgaaagt atcggaagtg acataacgtt ccatctcgcg 3600
taaacgcgtt tcacttgccg ccagttcacg gtcgacttca tccagcaatt cgctgctgga 3660
aggtagctgc tcaccaaagg ccatgttgtc cggcattgga tcaagcgcaa atgacaaaat 3720
gatgtaagca accagggtaa acagcgccag accgaagaaa atcgacagca ccaccaggat 3780
acgcaccagt tttaccggta catcaaaata gttggcaatc ccggcgcaga cgccgcggac 3840
catgccctgc tgtggaatac gccataattt tttattgaga ttaatgcccg ccattagcga 3900
tccctccagt tcggatgttc tgcatcaaga atagattcca gcgcctgaat acgttcgcgc 3960
atccgttttg cttcatcagc cagttgcgct aatcgctgct gctcactttg cgacaattca 4020
ctgcgaccag aacgattgct gtaatgcagc cataaccaga tcggtaaaac aaacagcaca 4080
aaaatggtta acggaatagc cagaaatagc gcgctcataa gtactcctta gacgatgagc 4140
ggcggcgcgt tcggacgccg cctggatgtt attattgatt gtcttgcttc attttggctt 4200
ttaattgtgc cagttgttcg ctgattgcat catcggcttt cagttcggca aactgatcgt 4260
ccagcgattt ttgtttaccg aagctgtggc tttctgcttc cgcttccatc tggtcaatac 4320
gacgttcgaa agattcgaaa cgagccattg cttcatccag tttgccactg tccagctgac 4380
gacgcacatc gcgcgacgag tttgccgcct gatgacgtaa catcaatgcc tgctggcgag 4440
cgcgtgtttc gctcaatttg ttttccagct cgccaatctc tttcttcatg cgtgccagcg 4500
tatcgtccac cagcgtcact tcatgttcca gggacttaat cagatcggtc agtttctgtt 4560
tttcaattaa cgctgcacgt gccagatcct ctctctcttt cagcagcgcc agttcggctt 4620
tttcctgcca ttcaacctca cgcgccgacg cttgttcaat acggcgagtc agctgtttct 4680
tttctgccaa cgcgcgcgcc gaagtagaac gtacttcaac cagtgtatct tccatctcct 4740
ggatcatcag acgaaccagt ttctgtggat cttccgcttt ctctaacaga gcgttgatgt 4800
tggcattcac gatgtcggca aagcgagaaa aaatacccat aatgttgtcc tcttgatttc 4860
tgcgttcagg attgtcctgc tgaactgtta atacaatttg cgtgccaatt ttttatcttt 4920
ttgatttata aagatctgat tgaagaatca acagcaacat gccaggatga gttagcgaat 4980
tacactaaca agtggcgaat ttcatcatgg cagaatacaa agataattta cttggtgagg 5040
cgaacagctt tctcgaagtg ctggaacagg tttcgcatct cgcaccgctg gacaaaccgg 5100
tgctcatcat cggcgaacgc ggcaccggta aagagctgat tgccagccgc ctgcattatc 5160
tctcctcccg ttggcaaggg ccgtttattt cccttaactg cgcggcgtta aatgaaaatc 5220
tgctggattc cgaactgttt ggtcacgaag cgggggcgtt taccggtgcg caaaaacgtc 5280
atccagggag atttgaacgt gccgacggcg gtacgctatt tcttgatgaa ctcgctacgg 5340
caccgatgat ggtgcaggag aaattattgc gcgtgattga gtacggtgaa ctggagcgcg 5400
ttggcggcag ccaaccattg caggtgaatg tgcggttggt atgcgcgacg aatgccgatc 5460
tcccggcgat ggtcaatgaa ggcacttttc gcgctgacct gctcgaccga ctggcttttg 5520
atgttgtaca actgccacca ctgcgcgagc gcgaaagcga cataatgttg atggcagaat 5580
actttgccat ccagatgtgt cgggaaatca agctgcctct gttcccgggg tttacggagc 5640
gcgccagaga aacattgctg aattatcgtt ggccgggaaa tattcgtgaa ttgaaaaacg 5700
tggtggaacg ttcagtgtat cgccacggca ccagcgatta tccgcttgat gacatcatta 5760
ttgatccctt taaacggcgt ccgcctgaag acgctatcgc cgtttcagaa accacctcgc 5820
ttccaacact gccgctggat ttacgtgagt ttcagatgca gcaggaa 5867
<210> 4
<211> 4129
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
aggaggaaga ggacatccgg tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt 60
cgttttagac ttagggaccc tttatgacaa cttgacggct acatgatgaa attcgccact 120
tgttagtgta attcgctaac tcatcctggc atgttgctgt tgattcttca atcagatctt 180
tataaatcaa aaagataaaa aattggcacg caaattgtat taacagttca gcaggacaat 240
cctgaattct cacacaggaa acagctatga gcactgcaat tacacgccag atcgttctcg 300
ctaccgcaac caccggtatg aaccagattg gtgcgcacta tgaaggccac aagatcattg 360
agattggtgc cgttgaagtg gtgaaccgtc gcctgacggg caataacttc catgtttatc 420
tcaaacccga tcggctggtg gatccggaag cctttggcgt acatggtatt gccgatgaat 480
ttttgctcga taagcccacg tttgccgaag tagccgatga gttcatggac tatattcgcg 540
gcgcggagtt ggtgatccat aacgcagcgt tcgatatcgg ctttatggac tacgagtttt 600
cgttgcttaa gcgcgatatt ccgaagacca atactttctg taaggtcacc gatagccttg 660
cggtggcgag gaaaatgttt cccggtaagc gcaacagcct cgatgcgtta tgtgctcgct 720
acgaaataga taacagtaaa cgaacgctgc acggggcatt actcgatgcc cagatccttg 780
cggaagttta tctggcgatg accggtggtc aaacgtcgat ggcttttgcg atggaaggag 840
agacacaaca gcaacaaggt gaagcaacaa ttcagcgcat tgtacgtcag gcaagtaagt 900
tacgcgttgt ttttgcgaca gatgaagaga ttgcagctca tgaagcccgt ctcgatctgg 960
tgcagaagaa aggcggaagt tgcctctggc gagcataatt taatatcagt aaaccggaca 1020
taacccatga agaaaaatcg cgcttttttg aagtgggcag ggggcaagta tcccctgctt 1080
gatgatatta aacggcattt gcccaagggc gaatgtctgg ttgagccttt tgtaggtgcc 1140
gggtcggtgt ttctcaacac cgacttttct cgttatatcc ttgccgatat caatagcgac 1200
ctgatcagtc tctataacat tgtgaagatg cgtactgatg agtacgtaca ggccgcacgc 1260
gagctgtttg ttcccgaaac aaattgcgcc gaggtttact atcagttccg cgaagagttc 1320
aacaaaagcc aggatccgtt ccgtcgggcg gtactgtttt tatatttgaa ccgctacggt 1380
tacaacggcc tgtgtcgtta caatctgcgc ggtgagttta acgtgccgtt cggccgctac 1440
aaaaaaccct atttcccgga agcagagttg tatcacttcg ctgaaaaagc gcagaatgcc 1500
tttttctatt gtgagtctta cgccgatagc atggcgcgcg cagatgatgc atccgtcgtc 1560
tattgcgatc cgccttatgc accgctgtct gcgaccgcca actttacggc gtatcacaca 1620
aacagtttta cgcttgaaca acaagcgcat ctggcggaga tcgccgaagg tctggttgag 1680
cgccatattc cagtgctgat ctccaatcac gatacgatgt taacgcgtga gtggtatcag 1740
cgcgcaaaat tgcatgtcgt caaagttcga cgcagtataa gcagcaacgg cggcacacgt 1800
aaaaaggtgg acgaactgct ggctttgtac aaaccaggag tcgtttcacc cgcgaaaaaa 1860
taattcagct aagacactgc actggattaa gatgaaaacg attgaagttg atgatgaact 1920
ctacagctat attgccagcc acactaagca tatcggcgag agcgcatccg acattttacg 1980
gcgtatgttg aaattttccg ccgcatcaca gcctgctgct ccggtgacga aagaggttcg 2040
cgttgcgtca cctgctatcg tcgaagcgaa gccggtcaaa acgattaaag acaaggttcg 2100
cgcaatgcgt gaacttctgc tttcggatga atacgcagag caaaagcgag cggtcaatcg 2160
ctttatgctg ctgttgtcta cactatattc tcttgacgcc caggcgtttg ccgaagcaac 2220
ggaatcgttg cacggtcgta cacgcgttta ctttgcggca gatgaacaaa cgctgctgaa 2280
aaatggtaat cagaccaagc cgaaacatgt gccaggcacg ccgtattggg tgatcaccaa 2340
caccaacacc ggccgtaaat gcagcatgat cgaacacatc atgcagtcga tgcaattccc 2400
ggcggaattg attgagaagg tttgcggaac tatctaaact taattaacgg cactcctcag 2460
ccaagtcaaa agcctccgac cggaggcttt tgactacatg cccatggcgt ttacgccccg 2520
ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca 2580
tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta 2640
taatatttgc ccatagtgaa aacgggggcg aagaagttgt ccatattggc cacgtttaaa 2700
tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac 2760
cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt 2820
agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc 2880
tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc 2940
attgccatac ggaactccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc 3000
ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga 3060
acggtctggt tataggtaca ttgagtaact gactgaaatg cctcaaaatg ttctttacga 3120
tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc 3180
ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta 3240
tggtgaaagt tggaacctct tacgtgccaa gccaaatagg ccgtcactcg gtcgctacgc 3300
tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt 3360
ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc 3420
cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg 3480
ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag 3540
atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt 3600
taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca 3660
cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac 3720
tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg 3780
aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt 3840
tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag 3900
gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac 3960
tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga 4020
ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct 4080
agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagccc 4129
<210> 5
<211> 6587
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
tggtttttgt cactgtatgt tattggcgtt tcctgcacct acgatgtttt tgaccaacag 60
tttgctaatt tctttacttc gttctttgct accggtgaac agggtacgcg ggtatttggc 120
tacgtaacga caatgggcga attacttaac gcctcgatta tgttctttgc gccactgatc 180
attaatcgca tcggtgggaa aaacgccctg ctgctggctg gcactattat gtctgtacgt 240
attattggct catcgttcgc cacctcagcg ctggaagtgg ttattctgaa aacgctgcat 300
atgtttgaag taccgttcct gctggtgggc tgctttaaat atattaccag ccagtttgaa 360
gtgcgttttt cagcgacgat ttatctggtc tgtttctgct tctttaagca actggcgatg 420
atttttatgt ctgtactggc gggcaatatg tatgaaagca tcggtttcca gggcgcttat 480
ctggtgctgg gtctggtggc gctgggcttc accttaattt ccgtgttcac gcttagcggc 540
cccggcccgc tttccctgct gcgtcgtcag gtgaatgaag tcgcttaatt ccgtcaatat 600
ttaccttccc tccctcaatc ggttgaatgt cgcccttttg tctttagcgc tggtaaacca 660
tatgaatttt ctattgattg tgacaaaata aacttattcc gtggtgtctt tgcgtttctt 720
ttatatgttg ccacctttat gtatgtattt tctacgtttg ctaacatact gcgtaataag 780
gagtcttaat catgccagtt cttttgggta ttccgttatt attgcgtttc ctcggtttcc 840
ttctggtaac tttgttcggc tatctgctta cttttcttaa aaagggcttc ggtaagatag 900
ctattgctat ttcattgttt cttgctctta ttattgggct taactcaatt cttgtgggtt 960
atctctctga tattagcgct caattaccct ctgactttgt tcagggtgtt cagttaattc 1020
tcccgtctaa tgcgcttccc tgtttttatg ttattctctc tgtaaaggct gctattttca 1080
tttttgacgt taaacaaaaa atcgtttctt atttggattg ggataaataa tatggctgtt 1140
tattttgtaa ctggcaaatt aggctctgga aagacgctcg ttagcgttgg taagattcag 1200
gataaaattg tagctgggtg caaaatagca actaatcttg atttaaggct tcaaaacctc 1260
ccgcaagtcg ggaggttcgc taaaacgcct cgcgttctta gaataccgga taagccttct 1320
atatctgatt tgcttgctat tgggcgcggt aatgattcct acgatgaaaa taaaaacggc 1380
ttgcttgttc tcgatgagtg cggtacttgg tttaataccc gttcttggaa tgataaggaa 1440
agacagccga ttattgattg gtttctacat gctcgtaaat taggatggga tattattttt 1500
cttgttcagg acttatctat tgttgataaa caggcgcgtt ctgcattagc tgaacatgtt 1560
gtttattgtc gtcgtctgga cagaattact ttaccttttg tcggtacttt atattctctt 1620
attactggct cgaaaatgcc tctgcctaaa ttacatgttg gcgttgttaa atatggcgat 1680
tctcaattaa gccctactgt tgagcgttgg ctttatactg gtaagaattt gtataacgca 1740
tatgatacta aacaggcttt ttctagtaat tatgattccg gtgtttattc ttatttaacg 1800
ccttatttat cacacggtcg gtatttcaaa ccattaaatt taggtcagaa gatgaaatta 1860
actaaaatat atttgaaaaa gttttctcgc gttctttgtc ttgcgattgg atttgcatca 1920
gcatttacat atagttatat aacccaacct aagccggagg ttaaaaaggt agtctctcag 1980
acctatgatt ttgataaatt cactattgac tcttctcagc gtcttaatct aagctatcgc 2040
tatgttttca aggattctaa gggaaaatta attaatagcg acgatttaca gaagcaaggt 2100
tattcactca catatattga tttatgtact gtttccatta aaaaaggtaa ttcaaatgaa 2160
attgttaaat gtaattaatt ttgttttctt gatgtttgtt tcatcatctt cttttgctca 2220
ggtaattgaa atgaataatt cgcctctgcg cgattttgta acttggtatt caaagcaatc 2280
aggcgaatcc gttattgttt ctcccgatgt aaaaggtact gttactgtat attcatctga 2340
cgttaaacct gaaaatctac gcaatttctt tatttctgtt ttacgtgcta ataattttga 2400
tatggttggt tcaattcctt ccataattca gaagtataat ccaaacaatc aggattatat 2460
tgatgaattg ccatcatctg ataatcagga atatgatgat aattccgctc cttctggtgg 2520
tttctttgtt ccgcaaaatg ataatgttac tcaaactttt aaaattaata acgttcgggc 2580
aaaggattta atacgagttg tcgaattgtt tgtaaagtct aatacttcta aatcctcaaa 2640
tgtattatct attgacggct ctaatctatt agttgttagt gcacctaaag atattttaga 2700
taaccttcct caattccttt ctactgttga tttgccaact gaccagatat tgattgaggg 2760
tttgatattt gaggttcagc aaggtgatgc tttagatttt tcatttgctg ctggctctca 2820
gcgtggcact gttgcaggcg gtgttaatac tgaccgcctc acctctgttt tatcttctgc 2880
tggtggttcg ttcggtattt ttaatggcga tgttttaggg ctatcagttc gcgcattaaa 2940
gactaatagc cattcaaaaa tattgtctgt gccacgtatt cttacgcttt caggtcagaa 3000
gggttctatc tctgttggcc agaatgtccc ttttattact ggtcgtgtga ctggtgaatc 3060
tgccaatgta aataatccat ttcagacgat tgagcgtcaa aatgtaggta tttccatgag 3120
cgtttttcct gttgcaatgg ctggcggtaa tattgttctg gatattacca gcaaggccga 3180
tagtttgagt tcttctactc aggcaagtga tgttattact aatcaaagaa gtattgctac 3240
aacggttaat ttgcgtgatg gacagactct tttactcggt ggcctcactg attataaaaa 3300
cacttctcaa gattctggcg taccgttcct gtctaaaatc cctttaatcg gcctcctgtt 3360
tagctcccgc tctgattcca acgaggaaag cacgttatac gtgctcgtca aagcaaccat 3420
agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 3480
ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 3540
ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 3600
ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg 3660
ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 3720
gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt 3780
tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 3840
ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aatatttgct tatacaatct 3900
tcctgttttt ggggcttttc tgattatcaa ccggggtaca tatgattgac atgctagttt 3960
tacgattacc gttcatcgat tctcttgttt gctccagact ctcaggcaat gacctgatag 4020
cctttgtaga cctctcaaaa atagctaccc tctccggcat gaatttatca gctagaacgg 4080
ttgaatatca tattgatggt gatttgactg tctccggcct ttctcaccct tttgaatctt 4140
tacctacaca ttactcaggc attgcattta aaatatatga gggttctaaa aatttttatc 4200
cttgcgttga aataaaggct tctcccgcaa aagtattaca gggtcataat gtttttggta 4260
caaccgattt agctttatgc tctgaggctt tattgcttaa ttttgctaat tctttgcctt 4320
gcctgtatga tttattggat gttaacgcta ctactattag tagaattgat gccacctttt 4380
cagctcgcgc cccaaatgaa aatatagcta aacaggttat tgaccatttg cgaaatgtat 4440
ctaatggtca aactaaatct actcgttcgc agaattggga atcaactgtt acatggaatg 4500
aaacttccag acaccgtact ttagttgcat atttaaaaca tgttgagcta cagcaccaga 4560
ttcagcaatt aagctctaag ccatccgcaa aaatgacctc ttatcaaaag gagcaattaa 4620
aggtactctc taatcctgac ctgttggagt ttgcttccgg tctggttcgc tttgaagctc 4680
gaattaaaac gcgatatttg aagtctttcg ggcttcctct taatcttttt gatgcaatcc 4740
gctttgcttc tgactataat agtcagggta aagacctgat ttttgattta tggtcattct 4800
cgttttctga actgtttaaa gcatttgagg gggattcaat gaatatttat gacgattccg 4860
cagtattgga cgctatccag tctaaacatt ttactattac cccctctggc aaaacttctt 4920
ttgcaaaagc ctctcgctat tttggttttt atcgtcgtct ggtaaacgag ggttatgata 4980
gtgttgctct tactatgcct cgtaattcct tttggcgtta tgtatctgca ttagttgaat 5040
gtggtattcc taaatctcaa ctgatgaatc tttctacctg taataatgtt gttccgttag 5100
ttcgttttat taacgtagat ttttcttccc aacgtcctga ctggtataat gagccagttc 5160
ttaaaatcgc ataaggtaat tcacaatgat taaagttgaa attaaaccat ctcaagccca 5220
atttactact cgttctggtg tttctcgtca gggcaagcct tattcactga atgagcagct 5280
ttgttacgtt gatttgggta atgaatatcc ggttcttgtc aagattactc ttgatgaagg 5340
tcagccagcc tatgcgcctg gtctgtacac cgttcatctg tcctctttca aagttggtca 5400
gttcggttcc cttatgattg accgtctgcg cctcgttccg gctaagtaac atggagcagg 5460
tcgcggattt cgacacaatt tatcaggcga tgatacaaat ctccgttgta ctttgtttcg 5520
cgcttggtat aatcgctggg ggtcaaagat gagtgtttta gtgtattctt tcgcctcttt 5580
cgttttaggt tggtgccttc gtagtggcat tacgtatttt acccgtttaa tggaaacttc 5640
ctcatgaaaa agtctttagt cctcaaagcc tctgtagccg ttgctaccct cgttccgatg 5700
ctgtctttcg ctgctgaggg tgacgatccc gcaaaagcgg cctttaactc cctgcaagcc 5760
tcagcgaccg aatatatcgg ttatgcgtgg gcgatggttg ttgtcattgt cggcgcaact 5820
atcggtatca agctgtttaa gaaattcacc tcgaaagcaa gctgataaac cgatacaatt 5880
aaaggctcct tttggagcct ttttttttgg agattttcaa catgtactat ttaaaaaaca 5940
caaacttttg gatgttcggt ttattctttt tcttttactt ttttatcatg ggagcctact 6000
tcccgttttt cccgatttgg ctacatgaca tcaaccatat cagcaaaagt gatacgggta 6060
ttatttttgc cgctatttct ctgttctcgc tattattcca accgctgttt ggtctgcttt 6120
ctgacaaact cgggctgcgc aaatacctgc tgtggattat taccggcatg ttagtgatgt 6180
ttgcgccgtt ctttattttt atcttcgggc cactgttaca atacaacatt ttagtaggat 6240
cgattgttgg tggtatttat ctaggctttt gttttaacgc cggtgcgcca gcagtagagg 6300
catttattga gaaagtcagc cgtcgcagta atttcgaatt tggtcgcgcg cggatgtttg 6360
gctgtgttgg ctgggcgctg tgtgcctcga ttgtcggcat catgttcacc atcaataatc 6420
agtttgtttt ctggctgggc tctggctgtg cactcatcct cgccgtttta ctctttttcg 6480
ccaaaacgga tgcgccctct tctgccacgg ttgccaatgc ggtaggtgcc aaccattcgg 6540
catttagcct taagctggca ctggaactgt tcagacagcc aaaactg 6587
<210> 6
<211> 14
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
tgggagcgct ccca 14
<210> 7
<211> 6114
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
cgtcaaagtg gccgaacttc ataagctttt tctcctcttt ctctaactag tgctagcaca 60
gtacctagga ctgagctagc tgtcaaggta cctttacagc tagctcagtc ctagggactg 120
tgctagcgaa ttctagagaa agaggagaaa ctcgagatgt actatttaaa aaacacaaac 180
ttttggatgt tcggtttatt ctttttcttt tactttttta tcatgggagc ctacttcccg 240
tttttcccga tttggctaca tgacatcaac catatcagca aaagtgatac gggtattatt 300
tttgccgcta tttctctgtt ctcgctatta ttccaaccgc tgtttggtct gctttctgac 360
aaactcgggc tgcgcaaata cctgctgtgg attattaccg gcatgttagt gatgtttgcg 420
ccgttcttta tttttatctt cgggccactg ttacaataca acattttagt aggatcgatt 480
gttggtggta tttatctagg cttttgtttt aacgccggtg cgccagcagt agaggcattt 540
attgagaaag tcagccgtcg cagtaatttc gaatttggtc gcgcgcggat gtttggctgt 600
gttggctggg cgctgtgtgc ctcgattgtc ggcatcatgt tcaccatcaa taatcagttt 660
gttttctggc tgggctctgg ctgtgcactc atcctcgccg ttttactctt tttcgccaaa 720
acggatgcgc cctcttctgc cacggttgcc aatgcggtag gtgccaacca ttcggcattt 780
agccttaagc tggcactgga actgttcaga cagccaaaac tgtggttttt gtcactgtat 840
gttattggcg tttcctgcac ctacgatgtt tttgaccaac agtttgctaa tttctttact 900
tcgttctttg ctaccggtga acagggtacg cgggtatttg gctacgtaac gacaatgggc 960
gaattactta acgcctcgat tatgttcttt gcgccactga tcattaatcg catcggtggg 1020
aaaaacgccc tgctgctggc tggcactatt atgtctgtac gtattattgg ctcatcgttc 1080
gccacctcag cgctggaagt ggttattctg aaaacgctgc atatgtttga agtaccgttc 1140
ctgctggtgg gctgctttaa atatattacc agccagtttg aagtgcgttt ttcagcgacg 1200
atttatctgg tctgtttctg cttctttaag caactggcga tgatttttat gtctgtactg 1260
gcgggcaata tgtatgaaag catcggtttc cagggcgctt atctggtgct gggtctggtg 1320
gcgctgggct tcaccttaat ttccgtgttc acgcttagcg gccccggccc gctttccctg 1380
ctgcgtcgtc aggtgaatga agtcgcttaa ctgcaggtcc ctaagtctcc tcagcaaaac 1440
gaaaggccca gtctttcgac tgagcctttc gttttatttg accggatgtc ctcttgttca 1500
tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg 1560
aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac 1620
atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac 1680
gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc 1740
agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 1800
acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 1860
gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg 1920
tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt 1980
gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct 2040
cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 2100
aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 2160
aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 2220
tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 2280
caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 2340
cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 2400
ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 2460
gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 2520
agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 2580
gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 2640
acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 2700
gaggtggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 2760
gcaagcagca gattacgcgc agaaaaaaag gatctcaaac ggcctatttg gcctattttt 2820
ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 2880
atattgaaaa aggaagagta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc 2940
agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta 3000
cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt 3060
gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc 3120
ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga 3180
cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg 3240
caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt 3300
gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt 3360
tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa 3420
ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg 3480
gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga 3540
gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca 3600
agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa 3660
aggcgagatc accaaggtag tcggcaaata aacgccatgg ttaacccagg gtcacctcaa 3720
cgttgcaaac ggtgcccgct tgctgcggcg gcacaatgta accgtcgatc gcgttgccgt 3780
tcagggtcag gcttttaaca cccttgctca cgtgggtcgg gttggtaaca atgatgttat 3840
aggtcgcacc acggtattta cgggtcacct taaagccgtc ccagctgctc ggaatgcacg 3900
ggttgatcgc cagaccgcta taatccggtt taacgcccag aatgtattgg gtgatcgccg 3960
caaagttcca gctcgcggtg ccggtcagcc agctgttttt accttcgccc ggcagatacg 4020
cgtctttacc cgcaatcatc tggcaatacg cgtacggctc caccttgtgc agatcgctaa 4080
tttcctccag gtacgccgga gcgattttac gatagtattc aaacgcctta tcaccacggc 4140
ccagcagggt ctcggtaatc atgatccacg ggttgttgtg gcagaagata cccgcgtttt 4200
ctttataacc cgccgggtag gtgctaatct cgccatattc gatatagtac ttggtgaacg 4260
ccggttgttg cagaacaatg ccgtagtcgc aatccagcca ctctttaacg ctgtccatgc 4320
tcttttccac cagaccatcc tccagaccga tacccgccat accgcaaaag ccctggcttt 4380
caatgaagat tttgccttcc tcgttctcct tgctacccac tttcttgcca tagtagtcgt 4440
acgcacgcag gaaccattca ccatcccagc cgtgtttctt aaccgcctcc accatttggt 4500
caatgtgcgc ctgcgcctcc gccgcttcac catcttggcc gatttcacgg cacagtttaa 4560
caaactcgtt accgtacagc acgaacagac ccgcaatcat caggctttcc gcggtacgac 4620
cggtcttgtt gccggtggtc tggaagctct cgttcgggtc ctcgctgaag cagttcaggt 4680
tcaggcagtc gttccaatcc gcacgaccga tcagcggcag accgtgcggg cccaggttgt 4740
taacggtgtg gtaaaagcta cgtttcaggt gttcgaagtg gctggtcgcc ttgctcgcgt 4800
cgttatcata cggcacctgt tcctccagga tgctaaagtc gccggtctct ttaatgtaat 4860
cggtggtgct caggatcagc cacagcggat cgtcgttgaa gttaccgcca atcgcgttgt 4920
taccacgctt ggtcagcggt tgatactggt ggtacgcgct gccgtcttca aactgggtgc 4980
tcgcgatgtc aatgatacgt tcacgcgcac gctccggaac ttggtgcaca aaaccaatca 5040
ggtcctggtt gctatcacgg aaacccatgc cacgaccgat gccgctctca aagaagctcg 5100
cgctacggct catgttgaag gtaaccatgc attggtactg gttccagatg ttcaccatac 5160
ggtccagttt ttcatcaccg ctctcaacgc tataggtgct cagcagattc gcccagtagt 5220
ccgccagttt ggtcagttcc gcgtcaacct tctccacgct atcaaaacgc gcaatcagct 5280
ctttcgccgg gctcttgttg atcacgcctt tgctttccca cttgttctcc ggcgcaactt 5340
caatataacc cagcacgaag atcaggtctt tgctttcgcc cggcgccagc tcaacttcca 5400
ggtagtggct cgcgatcggg ctccaaccgt gcgcctcgct gttacgcggt tcaccctcaa 5460
acaccgcatc cggcttgtcg aaatcgttat acagaccttt ccacttgtca cgatcggtgt 5520
cgaagccttg aatcggcgcg ttaacgctat aaaacgcgta gtggttacga cgctctttga 5580
actcggtctt gtggtagatc acgctgtcct caacttccac ctcaccggtg ctaaagttac 5640
gctggaagtt ggtcatatcg tcttccgcgt tccacaggca ccactccgca aagctgaaca 5700
gcttaaattt cttggtcgca ccgctggtgt ttttcaggct aactttttga atttccgccc 5760
aggtgcccag cgggatgaag ctcagcactt ccgcctgaat accgttacgc tcgccggtga 5820
tacgggtata gctcagaccg tgcgcgcagc tgtacgcatc cagctccgct ttaaccggct 5880
tccaacccgg gctccacacg tcaccgctat cgttgatata gaagtattta ccgccgttgt 5940
caaccggcac gttgttataa cggtaacggg tcagacgacg aaatttcgca tccttataga 6000
agctgtaacc gccaccggtg ttgctaacca ggctaaagaa gtcttcgtta cccagatagt 6060
taatccacgg gtacggggtc ttcggatcgg tgatcacgta ctcacgcgct ttat 6114
<210> 8
<211> 4740
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc 60
gatgagtggc agggcggggc gtaaacgcca tggttgacag ctagctcagt cctaggtata 120
atgctagctg tggaattctc tagagaaaga ggagaaatac tagatgatca ccaaagttac 180
cgcccgcaat aataaaatca ccccggttga actgctgaat cagaaattcg gcaataaaat 240
caatctgggc aatttcgccg atgccgtttt caccgatgcc gccttcaaaa atgttgccgg 300
catcgccaat ctgccgatga aagccccggt tatgcaggtt ctgatggaaa attgcatcgt 360
tagcaaatac ctgaaacagt tcgttccgga tcgcagcgtt tgcttcgttg aagaaggcca 420
gaaattctac atcgttctgg aagacgggca gaaaatcgag gtgccggaag atgttaataa 480
agccctgaaa gccaccgtta gcgatgttaa acattgggcc ggctacctga ccgaagatgg 540
cgaacatgtt atcgatctgc tgaaaccggc cccgggcccg catttctacg ttaatctgct 600
gatcggcaat cgcctgggct tcaaacgcac cctccagacc accccgaaaa gcgttgttga 660
tcgcttcggc cgcggcagct tccgcagcca tgccgccacc caggttctgg ccacccgctt 720
cgatatgcgc caggaagaaa atggcttccc ggccaatcgc cagttctacc tgtacgaaga 780
tggcaaacaa atcttctaca gcgccctgat cgatgataat atcgttgaag ccacctgcaa 840
acatagctgc aatcgcaccg ttatcaaata caaaaccgcc tgcaatctgg aaatcacccg 900
caccatcttc ctggttccgc ataaaaaagg cttcccgctg gccaccgaac tccagcgcat 960
cgaaatcaaa aatgccagcg ataaagcccg caatctgagc atcacctaca ccggcatgtt 1020
cggcaccggc gccgttcatg ccatcttcga agatgttacc tacaccaatg ttatcatgca 1080
gagcgccgcc ctgtacaatg ataaaggcga gttcatcggc atcaccccgg attactaccc 1140
ggaagagttc aaacaggata cccgcttcgt taccatgatc gttcgcaatg gcgatgagaa 1200
gagcttccca cagagcttct gcaccgatta caatgatttc gttggcaccg gcaccctgga 1260
acatccggcc ggcggctgca atctgaataa taaactgaat cgcaaaggcc cgggcttctt 1320
cgccctgggc gccccgttca ccgttgaacc gggcaaaacc gttatcatcg ataccttcac 1380
cggcctgagc agcagcaagg ataacgagaa ttacagcgat gccgttatgc tgcgcgaact 1440
ggataatctg ctgcgctact tcgaaaaaag cgagagcgta gaggaaaccc tgaatgaaat 1500
catcaatttc catgaaaatt acggcaaata cttccagttc aataccggca ataaactgtt 1560
cgatagcggc ttcaatcgca atctggcctt ccaggttctg taccagacct tcatgagccg 1620
cagcttcggc cagacccaga aaggctaccg cgaaatcggc ttccgcgaaa tccaggatct 1680
gttcgccagc atgtactact tcatcaatat cggctaccag gatttcgtta aagaactgct 1740
gttcgaatgg accgccaatg tttacaaaat gggctacgcc aatcataatt tctactgggt 1800
tggcaaacag ccgggcctgt acagcgatga tagcctgtgg ctgctccagg cctactaccg 1860
ctacatcatc tacaccaaag ataccagcgt tctgaatgaa gaagttccgg ttgccgatgg 1920
caataatgaa aaacgcgccg ttcgcgaaac tctgaaagca attatccagt acagcgcctg 1980
catcagcgtt ggcgatcatg gcctgccgct gctggatctg gccgattgga atgattgcct 2040
gaaaatcgcc agcaatagca tcgatggcgc caccaaagaa aaactgtact acgaacagct 2100
gaaaaaaacc aatggcaaat acggcgatcg cttcatgagc gattacagcg aaagcgttat 2160
gaatgccttc ctgctgaaac tggccatcga tcatctggcc gaaatcgcca ccctggataa 2220
tgatacccag ctggcccagc agatgagcga actgagcaaa gaagttaccg atcgcatcca 2280
gaaacatgcc tggaaagaaa atttcttcgc ccgcgttctg atcaatcgct acaaagatgg 2340
cagctacacc tacctgggcg ccaaaggcga taaactgagc gccgatccga atatcgatgg 2400
cgtttacttc ctgaatagct tcgcctggag cgttctgagc gatgttgcca ccgatgaaca 2460
gatcgccatc atggttgatg ttatcaaaaa acatctgctg accccgtacg gcctgcgcct 2520
ggttaccccg gccgatctga ataaaatcgc caatgatacc gccaccggcc attacttctt 2580
cggcgatcgc gaaaatggcg ccgttttcaa acatgccagc atgatggccg ttgccgccct 2640
gatcaaagcc gccaaaaaag ttaaagataa cgagctggcc aaagaaatgg cccgcatcgc 2700
ctacttcatg atcgatctgg ttctgccgta caaaaatctg gaaaatccgt tccaggttgc 2760
cggcaatccg cgcatctgca cccagtacat caataccgat accggcgaaa atatcggccc 2820
gctgctgagc ggcaccgcca cctggctgaa tctgaatctg atcagcctgg ccggcatcga 2880
atacacccgc gatggcatca gcttcaatcc gatcctgcgc gaagaagaaa cccagctgaa 2940
tttcacgctg aaggcgccga agtgcagcta caaattcagc atcaccaaac cggttggctt 3000
cgcccgcatg gaaagcagcg aatacgaact gttcgttgat ggccagaaaa tcgataatac 3060
cgttatcccg atgtacaccg atgaaaaaga acatatcgtt accctgaaat tcaaataact 3120
gcagagggtc cctaagtcta aaacgaaagg cccagtcttt cgactgagcc tttcgtttta 3180
tttgaccgga tgtcctcttc ctcctgggct gacttcaggt gctacatttg aagagataaa 3240
ttgcactgaa atctagagcg gttcagtaga aaagatcaaa ggatcttctt gagatccttt 3300
ttttctgcgc gtaatctttt gccctgtaaa cgaaaaaacc acctggggag gtggtttgat 3360
cgaaggttaa gtcagttggg gaactgctta accgtggtaa ctggctttcg cagagcacag 3420
caaccaaatc tgtccttcca gtgtagccgg actttggcgc acacttcaag agcaaccgcg 3480
tgtttagcta aacaaatcct ctgcgaactc ccagttacca atggctgctg ccagtggcgt 3540
tttaccgtgc ttttccgggt tggactcaag tgaacagtta ccggataagg cgcagcagtc 3600
gggctgaacg gggagttctt gcttacagcc cagcttggag cgaacgacct acaccgagcc 3660
gagataccag tgtgtgagct atgagaaagc gccacacttc ccgtaaggga gaaaggcgga 3720
acaggtatcc ggtaaacggc agggtcggaa caggagagcg caagagggag cgacccgccg 3780
gaaacggtgg ggatctttaa gtcctgtcgg gtttcgcccg tactgtcaga ttcatggttg 3840
agcctcacgg ctcccacaga tgcaccggaa aagcgtctgt ttatgtgaac tctggcagga 3900
gggcggagcc tatggaaaaa cgccaccggc gcggccctgc tgttttgcct cacatgttag 3960
tcccctgctt atccacggaa tctgtgggta actttgtatg tgtccgcagc gcccgccgca 4020
gtctcacgcc cggagcgtag cgaccgagtg acggcctatt tggcttggca cgtaagaggt 4080
tccaactttc accataatga aataagatca ctaccgggcg tattttttga gttatcgaga 4140
ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat 4200
atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttac tcaatgtacc 4260
tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag 4320
cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggag 4380
ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac 4440
accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat 4500
ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc 4560
tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt 4620
ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcact 4680
atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat 4740
Claims (2)
1.一种高效转运纤维寡糖的LacY突变体，其特征在于：所述LacY突变体为对野生型的LacY进行以下位点突变：A177V/V197A，所述野生型的LacY氨基酸序列如SEQID NO.1所示。
2.权利要求1所述的LacY突变体在纤维素发酵中的应用。
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109943581A (zh) * | 2017-12-20 | 2019-06-28 | 深圳先进技术研究院 | 一种质粒以及噬菌体辅助的连续定向进化系统和定向进化方法 |
| CN111269294A (zh) * | 2018-12-05 | 2020-06-12 | 广州中国科学院先进技术研究所 | 纤维寡糖转运蛋白LacY的突变位点、突变转运蛋白LacY及其制备方法和应用 |
| CN112239489A (zh) * | 2020-10-20 | 2021-01-19 | 广西科学院 | 一种海栖热袍菌纤维寡糖转运蛋白、其编码基因及应用 |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109943581A (zh) * | 2017-12-20 | 2019-06-28 | 深圳先进技术研究院 | 一种质粒以及噬菌体辅助的连续定向进化系统和定向进化方法 |
| CN111269294A (zh) * | 2018-12-05 | 2020-06-12 | 广州中国科学院先进技术研究所 | 纤维寡糖转运蛋白LacY的突变位点、突变转运蛋白LacY及其制备方法和应用 |
| CN112239489A (zh) * | 2020-10-20 | 2021-01-19 | 广西科学院 | 一种海栖热袍菌纤维寡糖转运蛋白、其编码基因及应用 |
Non-Patent Citations (2)
| Title |
|---|
| Enhancing phage display of antibody fragments using gIII-amber suppression;Mi-young Oh 等;Gene;第386卷(第1-2期);81-89 * |
| 嗜碱芽孢杆菌N16-5木寡糖结合蛋白XynE的功能表征;张周刚等;微生物学报;第55卷(第1期);40-49 * |
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