|
实施例5 |
实施例6 |
实施例7 |
实施例8 |
实施例9 |
蛋白浓度(质量%) |
19.5 |
20.0 |
20.5 |
21.5 |
22.0 |
有无在干燥室内制备 |
有 |
有 |
有 |
有 |
无 |
有无在干燥室内保管 |
有 |
有 |
有 |
有 |
无 |
如图2所示,实施例9(在未进行湿度管理的环境中制造、保存的蛋白浓度为22.0质量%)的纺丝原液的温度与粘度的关系和实施例7(在干燥室中制造、保存的蛋白浓度为20.5质量%)的纺丝原液的温度与粘度的关系大致相同。认为其原因在于,在对实施例9的纺丝原液进行制造、保存的期间,大气中的水分混入到实施例9的纺丝原液中。由这些结果确认了,在纺丝原液的制造时或保存时,通过降低环境气体的相对湿度、抑制纺丝原液的吸水,可以高度地维持纺丝原液的粘度。
(参考试验1)
对上述获得的蛛丝蛋白(粉体)进行真空干燥(全干)之后,将全干状态的蛛丝蛋白(粉体)暴露在温度为25℃、相对湿度为72%RH的大气中,研究含水率的变化。将其结果示于图3中。由图3可知,全干状态的蛛丝蛋白(粉体)约20分钟时水分率达到约13质量%(平衡水分率)。由此可知,对粉体的蛛丝蛋白进行真空干燥之后制成纺丝原液是重要的。在未进行真空干燥时,在室温或相对湿度高的环境下保存的蛛丝蛋白的粉体中所吸附的水分直接进入到纺丝原液中。
(参考试验2)
为了确认因水分混入到纺丝原液中所导致的纺丝原液的粘度降低是否是仅因纺丝原液的水分而发生稀释所导致的,进行以下的试验。首先,准备温度50℃下的粘度与实施例1的纺丝原液为同等程度的餐具用洗涤剂。接着,按照以所准备的餐具用洗涤剂为100质量%时的餐具用洗涤剂中的水含量达到3质量%的方式,在餐具用洗涤剂中添加水分,在温度50℃下测定粘度,研究水分添加前和添加后的粘度的变化率。
其结果,添加水分前的餐具用洗涤剂在温度50℃下的粘度为143mPa·s、水分添加后的餐具用洗涤剂在温度50℃下的粘度为133mPa·s。餐具用洗涤剂因水分添加而导致温度50℃下的粘度的降低率达到7%。与此相对,实施例1的纺丝原液在温度50℃下的粘度为153mPa·s,通过水分添加而水含量达到3质量%的实施例4的纺丝原液在温度50℃下的粘度为89mPa·s。溶解有蛛丝蛋白的纺丝原液因水分添加导致的温度50℃下的粘度降低率为42%。由此清楚可知,在溶解蛛丝蛋白等多肽而成的极性溶剂溶液中因水分混入所导致的粘度降低并非是仅因水分而发生稀释所导致的。
多肽的极性溶剂溶液的粘度因水分在溶液中的混入(含有)而极端地降低认为是由于以下的理由。构成蛋白(多肽)分子的氨基酸具有多种侧链。因此,水分子进入到蛋白的极性溶剂溶液中时,在蛋白分子的侧链之间形成氢键,蛋白分子发生凝聚。由此,认为蛋白的溶解性降低、蛋白的极性溶剂溶液的粘度降低。因而,相反若从蛋白的极性溶剂溶液中将水分子除去,则可以抑制蛋白分子的凝聚、提高蛋白在极性溶剂溶液中的溶解性,其结果可以提高极性溶剂溶液的粘度。
产业上的可利用性
本发明的极性溶剂溶液在湿式纺纱、膜流延、凝胶、颗粒、网状物、各种成型物中是有用的。
序列表自由文本
序列号1~4、12、13 氨基酸序列
序列号5~7 碱基序列
序列号8~11 引物序列
序 列 表
<110> 丝芭博株式会社
<120> 极性溶剂溶液及其制造方法
<130> H4382
<150> JP 2015-80226
<151> 2015-04-09
<160> 13
<170> PatentIn version 3.3
<210> 1
<211> 1154
<212> PRT
<213> Artificial
<220>
<223> ADF3Kai-Large-NRSH1
<400> 1
Met His His His His His His His His His His Ser Ser Gly Ser Ser
1 5 10 15
Leu Glu Val Leu Phe Gln Gly Pro Ala Arg Ala Gly Ser Gly Gln Gln
20 25 30
Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly
35 40 45
Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr
50 55 60
Gly Pro Gly Ser Gly Gln Gln Gly Pro Ser Gln Gln Gly Pro Gly Gln
65 70 75 80
Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala
85 90 95
Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro
100 105 110
Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ser Ser Ala Ala Ala Ala Ala
115 120 125
Ala Gly Gly Asn Gly Pro Gly Ser Gly Gln Gln Gly Ala Gly Gln Gln
130 135 140
Gly Pro Gly Gln Gln Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala
145 150 155 160
Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly
165 170 175
Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala
180 185 190
Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gly Pro Gly Gln Gln
195 200 205
Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala
210 215 220
Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly
225 230 235 240
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly
245 250 255
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
260 265 270
Tyr Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro
275 280 285
Tyr Gly Pro Gly Ala Ser Ala Ala Ser Ala Ala Ser Gly Gly Tyr Gly
290 295 300
Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln
305 310 315 320
Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly
325 330 335
Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
340 345 350
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly
355 360 365
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
370 375 380
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
385 390 395 400
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
405 410 415
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly
420 425 430
Gln Gly Ala Tyr Gly Pro Gly Ala Ser Ala Ala Ala Gly Ala Ala Gly
435 440 445
Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
450 455 460
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
465 470 475 480
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Tyr Gly
485 490 495
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
500 505 510
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
515 520 525
Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ala Ser Ala Ala Val Ser
530 535 540
Val Ser Arg Ala Arg Ala Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln
545 550 555 560
Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Tyr Gly Pro Gly
565 570 575
Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly
580 585 590
Gln Gln Gly Pro Ser Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly
595 600 605
Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly
610 615 620
Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro
625 630 635 640
Tyr Gly Pro Gly Ser Ser Ala Ala Ala Ala Ala Ala Gly Gly Asn Gly
645 650 655
Pro Gly Ser Gly Gln Gln Gly Ala Gly Gln Gln Gly Pro Gly Gln Gln
660 665 670
Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro
675 680 685
Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly
690 695 700
Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr
705 710 715 720
Gly Pro Gly Ser Gly Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln
725 730 735
Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly
740 745 750
Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
755 760 765
Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala
770 775 780
Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Tyr Gly Gln Gln Gly
785 790 795 800
Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala
805 810 815
Ser Ala Ala Ser Ala Ala Ser Gly Gly Tyr Gly Pro Gly Ser Gly Gln
820 825 830
Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro
835 840 845
Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser
850 855 860
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
865 870 875 880
Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala
885 890 895
Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly
900 905 910
Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
915 920 925
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
930 935 940
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Ala Tyr Gly
945 950 955 960
Pro Gly Ala Ser Ala Ala Ala Gly Ala Ala Gly Gly Tyr Gly Pro Gly
965 970 975
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
980 985 990
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
995 1000 1005
Gln Gln Gly Pro Gly Gln Gln Gly Pro Tyr Gly Pro Gly Ala Ser
1010 1015 1020
Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln
1025 1030 1035
Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly
1040 1045 1050
Gln Gly Pro Tyr Gly Pro Gly Ala Ala Ser Ala Ala Val Ser Val
1055 1060 1065
Gly Gly Tyr Gly Pro Gln Ser Ser Ser Val Pro Val Ala Ser Ala
1070 1075 1080
Val Ala Ser Arg Leu Ser Ser Pro Ala Ala Ser Ser Arg Val Ser
1085 1090 1095
Ser Ala Val Ser Ser Leu Val Ser Ser Gly Pro Thr Lys His Ala
1100 1105 1110
Ala Leu Ser Asn Thr Ile Ser Ser Val Val Ser Gln Val Ser Ala
1115 1120 1125
Ser Asn Pro Gly Leu Ser Gly Cys Asp Val Leu Val Gln Ala Leu
1130 1135 1140
Leu Glu Val Val Ser Ala Leu Val Ser Ile Leu
1145 1150
<210> 2
<211> 660
<212> PRT
<213> Artificial
<220>
<223> ADF3Kai
<400> 2
Met His His His His His His His His His His Ser Ser Gly Ser Ser
1 5 10 15
Leu Glu Val Leu Phe Gln Gly Pro Ala Arg Ala Gly Ser Gly Gln Gln
20 25 30
Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly
35 40 45
Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr
50 55 60
Gly Pro Gly Ser Gly Gln Gln Gly Pro Ser Gln Gln Gly Pro Gly Gln
65 70 75 80
Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala
85 90 95
Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro
100 105 110
Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ser Ser Ala Ala Ala Ala Ala
115 120 125
Ala Gly Gly Asn Gly Pro Gly Ser Gly Gln Gln Gly Ala Gly Gln Gln
130 135 140
Gly Pro Gly Gln Gln Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala
145 150 155 160
Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly
165 170 175
Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala
180 185 190
Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gly Pro Gly Gln Gln
195 200 205
Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala
210 215 220
Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly
225 230 235 240
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly
245 250 255
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
260 265 270
Tyr Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro
275 280 285
Tyr Gly Pro Gly Ala Ser Ala Ala Ser Ala Ala Ser Gly Gly Tyr Gly
290 295 300
Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln
305 310 315 320
Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly
325 330 335
Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
340 345 350
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly
355 360 365
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
370 375 380
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
385 390 395 400
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
405 410 415
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly
420 425 430
Gln Gly Ala Tyr Gly Pro Gly Ala Ser Ala Ala Ala Gly Ala Ala Gly
435 440 445
Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
450 455 460
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
465 470 475 480
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Tyr Gly
485 490 495
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
500 505 510
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
515 520 525
Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ala Ser Ala Ala Val Ser
530 535 540
Val Gly Gly Tyr Gly Pro Gln Ser Ser Ser Val Pro Val Ala Ser Ala
545 550 555 560
Val Ala Ser Arg Leu Ser Ser Pro Ala Ala Ser Ser Arg Val Ser Ser
565 570 575
Ala Val Ser Ser Leu Val Ser Ser Gly Pro Thr Lys His Ala Ala Leu
580 585 590
Ser Asn Thr Ile Ser Ser Val Val Ser Gln Val Ser Ala Ser Asn Pro
595 600 605
Gly Leu Ser Gly Cys Asp Val Leu Val Gln Ala Leu Leu Glu Val Val
610 615 620
Ser Ala Leu Val Ser Ile Leu Gly Ser Ser Ser Ile Gly Gln Ile Asn
625 630 635 640
Tyr Gly Ala Ser Ala Gln Tyr Thr Gln Met Val Gly Gln Ser Val Ala
645 650 655
Gln Ala Leu Ala
660
<210> 3
<211> 1183
<212> PRT
<213> Artificial
<220>
<223> ADF3Kai-Large
<400> 3
Met His His His His His His His His His His Ser Ser Gly Ser Ser
1 5 10 15
Leu Glu Val Leu Phe Gln Gly Pro Ala Arg Ala Gly Ser Gly Gln Gln
20 25 30
Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly
35 40 45
Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr
50 55 60
Gly Pro Gly Ser Gly Gln Gln Gly Pro Ser Gln Gln Gly Pro Gly Gln
65 70 75 80
Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala
85 90 95
Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro
100 105 110
Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ser Ser Ala Ala Ala Ala Ala
115 120 125
Ala Gly Gly Asn Gly Pro Gly Ser Gly Gln Gln Gly Ala Gly Gln Gln
130 135 140
Gly Pro Gly Gln Gln Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala
145 150 155 160
Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly
165 170 175
Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala
180 185 190
Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gly Pro Gly Gln Gln
195 200 205
Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala
210 215 220
Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly
225 230 235 240
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly
245 250 255
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
260 265 270
Tyr Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro
275 280 285
Tyr Gly Pro Gly Ala Ser Ala Ala Ser Ala Ala Ser Gly Gly Tyr Gly
290 295 300
Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln
305 310 315 320
Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly
325 330 335
Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
340 345 350
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly
355 360 365
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
370 375 380
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
385 390 395 400
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
405 410 415
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly
420 425 430
Gln Gly Ala Tyr Gly Pro Gly Ala Ser Ala Ala Ala Gly Ala Ala Gly
435 440 445
Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
450 455 460
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
465 470 475 480
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Tyr Gly
485 490 495
Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly
500 505 510
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
515 520 525
Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ala Ser Ala Ala Val Ser
530 535 540
Val Ser Arg Ala Arg Ala Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln
545 550 555 560
Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Tyr Gly Pro Gly
565 570 575
Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly
580 585 590
Gln Gln Gly Pro Ser Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly
595 600 605
Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly
610 615 620
Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro
625 630 635 640
Tyr Gly Pro Gly Ser Ser Ala Ala Ala Ala Ala Ala Gly Gly Asn Gly
645 650 655
Pro Gly Ser Gly Gln Gln Gly Ala Gly Gln Gln Gly Pro Gly Gln Gln
660 665 670
Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro
675 680 685
Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly
690 695 700
Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr
705 710 715 720
Gly Pro Gly Ser Gly Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln
725 730 735
Gly Pro Tyr Gly Pro Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly
740 745 750
Tyr Gly Pro Gly Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
755 760 765
Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala
770 775 780
Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Tyr Gly Gln Gln Gly
785 790 795 800
Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala
805 810 815
Ser Ala Ala Ser Ala Ala Ser Gly Gly Tyr Gly Pro Gly Ser Gly Gln
820 825 830
Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro
835 840 845
Gly Ala Ser Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser
850 855 860
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
865 870 875 880
Gln Gln Gly Pro Gly Gly Gln Gly Pro Tyr Gly Pro Gly Ala Ser Ala
885 890 895
Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln Gln Gly
900 905 910
Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
915 920 925
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
930 935 940
Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly Gln Gly Ala Tyr Gly
945 950 955 960
Pro Gly Ala Ser Ala Ala Ala Gly Ala Ala Gly Gly Tyr Gly Pro Gly
965 970 975
Ser Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro
980 985 990
Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly
995 1000 1005
Gln Gln Gly Pro Gly Gln Gln Gly Pro Tyr Gly Pro Gly Ala Ser
1010 1015 1020
Ala Ala Ala Ala Ala Ala Gly Gly Tyr Gly Pro Gly Ser Gly Gln
1025 1030 1035
Gln Gly Pro Gly Gln Gln Gly Pro Gly Gln Gln Gly Pro Gly Gly
1040 1045 1050
Gln Gly Pro Tyr Gly Pro Gly Ala Ala Ser Ala Ala Val Ser Val
1055 1060 1065
Gly Gly Tyr Gly Pro Gln Ser Ser Ser Val Pro Val Ala Ser Ala
1070 1075 1080
Val Ala Ser Arg Leu Ser Ser Pro Ala Ala Ser Ser Arg Val Ser
1085 1090 1095
Ser Ala Val Ser Ser Leu Val Ser Ser Gly Pro Thr Lys His Ala
1100 1105 1110
Ala Leu Ser Asn Thr Ile Ser Ser Val Val Ser Gln Val Ser Ala
1115 1120 1125
Ser Asn Pro Gly Leu Ser Gly Cys Asp Val Leu Val Gln Ala Leu
1130 1135 1140
Leu Glu Val Val Ser Ala Leu Val Ser Ile Leu Gly Ser Ser Ser
1145 1150 1155
Ile Gly Gln Ile Asn Tyr Gly Ala Ser Ala Gln Tyr Thr Gln Met
1160 1165 1170
Val Gly Gln Ser Val Ala Gln Ala Leu Ala
1175 1180
<210> 4
<211> 24
<212> PRT
<213> Artificial
<220>
<223> His tag and start codon
<400> 4
Met His His His His His His His His His His Ser Ser Gly Ser Ser
1 5 10 15
Leu Glu Val Leu Phe Gln Gly Pro
20
<210> 5
<211> 1983
<212> DNA
<213> Artificial
<220>
<223> ADF3Kai
<400> 5
atgcatcacc atcatcatca tcaccaccac cattcctcgg gctcatcctt ggaagtgtta 60
tttcaaggac cagcacgagc cggttcggga caacaagggc ctggccagca gggcccaggt 120
caacaagggc caggacagca gggtccttat gggcccggcg caagcgcagc agctgcggcc 180
gctggtggct atggtcctgg ctccggtcaa cagggccctt cgcaacaagg tcccgggcag 240
caaggtcctg gtggccaggg tccctacggg ccgggggcga gtgcggcagc agccgctgca 300
ggcggttatg gtccaggaag cggacagcaa ggtccgggag gtcaaggtcc gtatggccca 360
ggctctagcg cggctgccgc tgccgcgggt ggcaacggac cagggagcgg acaacagggc 420
gcgggacaac agggtccagg acagcaaggc ccaggggcgt cggcggctgc agcggcggcc 480
ggaggctatg gacccggctc aggacaacag ggaccgggtc aacaaggacc cggtggccaa 540
ggcccctatg gcccgggcgc cagcgcggcc gcagccgccg cgggcgggta cggccccggt 600
agcggccagg gaccaggtca gcaggggcca ggaggtcagg gcccatacgg tccgggcgca 660
tccgcggcgg cggcagcggc aggtggctac ggtcccggaa gcggccaaca ggggccaggg 720
caacaaggac caggacaaca aggtcctggg ggccaaggac cgtatggacc aggagcatca 780
gctgcagccg cggcagctgg cggttacggt ccaggctacg gccagcaggg tccgggtcag 840
cagggaccgg gaggccaggg gccttatggc cctggcgctt ccgcagccag tgccgcttct 900
ggaggatacg ggccgggaag cggtcagcaa ggccctggcc aacaaggacc tggaggccaa 960
gggccctacg gcccaggagc ctcggcagcc gcagctgccg caggtgggta tgggccaggt 1020
agcgggcaac aagggccggg tcagcaagga ccggggcaac agggacctgg gcagcaagga 1080
cccgggggtc aaggcccgta cggacctggt gcgtctgcag ctgctgctgc ggctggtgga 1140
tatggtccgg gatcggggca gcagggtccc ggtcagcagg gccctggtca gcaagggcca 1200
ggccaacagg gacccggaca acaaggcccg ggtcaacagg gtcctggaca gcaggggccg 1260
ggccaacaag gccctgggca acagggtccg gggggacagg gggcctatgg gcctggcgca 1320
tctgccgccg ctggcgcagc cggtgggtac gggcctgggt caggtcaaca ggggcctggt 1380
caacaaggcc ccgggcaaca gggccccggc cagcaaggtc cagggcagca gggcccggga 1440
cagcaagggc ctggacaaca ggggcccgga cagcagggac cttacgggcc cggtgcgagc 1500
gcagcggccg ccgccgcagg gggatatggc cccggatcgg gccagcaggg accaggccag 1560
caaggacctg gccaacaggg cccggggggt caggggccgt atggtcccgg cgctgcaagt 1620
gctgcagtgt ccgttggagg ttacggccct cagtcttcgt ctgttccggt ggcgtccgca 1680
gttgcgagta gactgtcttc acctgctgct tcatcgcgag tatcgagcgc tgtttcgtct 1740
cttgtctcgt cgggtcccac gaaacatgcc gccctttcaa atacgatttc atctgtagtg 1800
tcccaagtta gtgcaagtaa cccggggtta tccggatgcg acgttctcgt tcaggcactc 1860
ctagaagtag tatccgcgtt ggtgagcatc ttaggcagct cctcgatagg tcaaataaac 1920
tatggtgctt cagcccagta tacacagatg gtgggacaga gcgtcgcgca ggcattggct 1980
taa 1983
<210> 6
<211> 3552
<212> DNA
<213> Artificial
<220>
<223> ADF3Kai-Large
<400> 6
atgcatcacc atcatcatca tcaccaccac cattcctcgg gctcatcctt ggaagtgtta 60
tttcaaggac cagcacgagc cggttcggga caacaagggc ctggccagca gggcccaggt 120
caacaagggc caggacagca gggtccttat gggcccggcg caagcgcagc agctgcggcc 180
gctggtggct atggtcctgg ctccggtcaa cagggccctt cgcaacaagg tcccgggcag 240
caaggtcctg gtggccaggg tccctacggg ccgggggcga gtgcggcagc agccgctgca 300
ggcggttatg gtccaggaag cggacagcaa ggtccgggag gtcaaggtcc gtatggccca 360
ggctctagcg cggctgccgc tgccgcgggt ggcaacggac cagggagcgg acaacagggc 420
gcgggacaac agggtccagg acagcaaggc ccaggggcgt cggcggctgc agcggcggcc 480
ggaggctatg gacccggctc aggacaacag ggaccgggtc aacaaggacc cggtggccaa 540
ggcccctatg gcccgggcgc cagcgcggcc gcagccgccg cgggcgggta cggccccggt 600
agcggccagg gaccaggtca gcaggggcca ggaggtcagg gcccatacgg tccgggcgca 660
tccgcggcgg cggcagcggc aggtggctac ggtcccggaa gcggccaaca ggggccaggg 720
caacaaggac caggacaaca aggtcctggg ggccaaggac cgtatggacc aggagcatca 780
gctgcagccg cggcagctgg cggttacggt ccaggctacg gccagcaggg tccgggtcag 840
cagggaccgg gaggccaggg gccttatggc cctggcgctt ccgcagccag tgccgcttct 900
ggaggatacg ggccgggaag cggtcagcaa ggccctggcc aacaaggacc tggaggccaa 960
gggccctacg gcccaggagc ctcggcagcc gcagctgccg caggtgggta tgggccaggt 1020
agcgggcaac aagggccggg tcagcaagga ccggggcaac agggacctgg gcagcaagga 1080
cccgggggtc aaggcccgta cggacctggt gcgtctgcag ctgctgctgc ggctggtgga 1140
tatggtccgg gatcggggca gcagggtccc ggtcagcagg gccctggtca gcaagggcca 1200
ggccaacagg gacccggaca acaaggcccg ggtcaacagg gtcctggaca gcaggggccg 1260
ggccaacaag gccctgggca acagggtccg gggggacagg gggcctatgg gcctggcgca 1320
tctgccgccg ctggcgcagc cggtgggtac gggcctgggt caggtcaaca ggggcctggt 1380
caacaaggcc ccgggcaaca gggccccggc cagcaaggtc cagggcagca gggcccggga 1440
cagcaagggc ctggacaaca ggggcccgga cagcagggac cttacgggcc cggtgcgagc 1500
gcagcggccg ccgccgcagg gggatatggc cccggatcgg gccagcaggg accaggccag 1560
caaggacctg gccaacaggg cccggggggt caggggccgt atggtcccgg cgctgcaagt 1620
gctgcagtgt ccgtttctag agcacgagcc ggttcgggac aacaagggcc tggccagcag 1680
ggcccaggtc aacaagggcc aggacagcag ggtccttatg ggcccggcgc aagcgcagca 1740
gctgcggccg ctggtggcta tggtcctggc tccggtcaac agggcccttc gcaacaaggt 1800
cccgggcagc aaggtcctgg tggccagggt ccctacgggc cgggggcgag tgcggcagca 1860
gccgctgcag gcggttatgg tccaggaagc ggacagcaag gtccgggagg tcaaggtccg 1920
tatggcccag gctctagcgc ggctgccgct gccgcgggtg gcaacggacc agggagcgga 1980
caacagggcg cgggacaaca gggtccagga cagcaaggcc caggggcgtc ggcggctgca 2040
gcggcggccg gaggctatgg acccggctca ggacaacagg gaccgggtca acaaggaccc 2100
ggtggccaag gcccctatgg cccgggcgcc agcgcggccg cagccgccgc gggcgggtac 2160
ggccccggta gcggccaggg accaggtcag caggggccag gaggtcaggg cccatacggt 2220
ccgggcgcat ccgcggcggc ggcagcggca ggtggctacg gtcccggaag cggccaacag 2280
gggccagggc aacaaggacc aggacaacaa ggtcctgggg gccaaggacc gtatggacca 2340
ggagcatcag ctgcagccgc ggcagctggc ggttacggtc caggctacgg ccagcagggt 2400
ccgggtcagc agggaccggg aggccagggg ccttatggcc ctggcgcttc cgcagccagt 2460
gccgcttctg gaggatacgg gccgggaagc ggtcagcaag gccctggcca acaaggacct 2520
ggaggccaag ggccctacgg cccaggagcc tcggcagccg cagctgccgc aggtgggtat 2580
gggccaggta gcgggcaaca agggccgggt cagcaaggac cggggcaaca gggacctggg 2640
cagcaaggac ccgggggtca aggcccgtac ggacctggtg cgtctgcagc tgctgctgcg 2700
gctggtggat atggtccggg atcggggcag cagggtcccg gtcagcaggg ccctggtcag 2760
caagggccag gccaacaggg acccggacaa caaggcccgg gtcaacaggg tcctggacag 2820
caggggccgg gccaacaagg ccctgggcaa cagggtccgg ggggacaggg ggcctatggg 2880
cctggcgcat ctgccgccgc tggcgcagcc ggtgggtacg ggcctgggtc aggtcaacag 2940
gggcctggtc aacaaggccc cgggcaacag ggccccggcc agcaaggtcc agggcagcag 3000
ggcccgggac agcaagggcc tggacaacag gggcccggac agcagggacc ttacgggccc 3060
ggtgcgagcg cagcggccgc cgccgcaggg ggatatggcc ccggatcggg ccagcaggga 3120
ccaggccagc aaggacctgg ccaacagggc ccggggggtc aggggccgta tggtcccggc 3180
gctgcaagtg ctgcagtgtc cgttggaggt tacggccctc agtcttcgtc tgttccggtg 3240
gcgtccgcag ttgcgagtag actgtcttca cctgctgctt catcgcgagt atcgagcgct 3300
gtttcgtctc ttgtctcgtc gggtcccacg aaacatgccg ccctttcaaa tacgatttca 3360
tctgtagtgt cccaagttag tgcaagtaac ccggggttat ccggatgcga cgttctcgtt 3420
caggcactcc tagaagtagt atccgcgttg gtgagcatct taggcagctc ctcgataggt 3480
caaataaact atggtgcttc agcccagtat acacagatgg tgggacagag cgtcgcgcag 3540
gcattggctt aa 3552
<210> 7
<211> 3465
<212> DNA
<213> Artificial
<220>
<223> ADF3Kai-Large-NRSH1
<400> 7
atgcatcacc atcatcatca tcaccaccac cattcctcgg gctcatcctt ggaagtgtta 60
tttcaaggac cagcacgagc cggttcggga caacaagggc ctggccagca gggcccaggt 120
caacaagggc caggacagca gggtccttat gggcccggcg caagcgcagc agctgcggcc 180
gctggtggct atggtcctgg ctccggtcaa cagggccctt cgcaacaagg tcccgggcag 240
caaggtcctg gtggccaggg tccctacggg ccgggggcga gtgcggcagc agccgctgca 300
ggcggttatg gtccaggaag cggacagcaa ggtccgggag gtcaaggtcc gtatggccca 360
ggctctagcg cggctgccgc tgccgcgggt ggcaacggac cagggagcgg acaacagggc 420
gcgggacaac agggtccagg acagcaaggc ccaggggcgt cggcggctgc agcggcggcc 480
ggaggctatg gacccggctc aggacaacag ggaccgggtc aacaaggacc cggtggccaa 540
ggcccctatg gcccgggcgc cagcgcggcc gcagccgccg cgggcgggta cggccccggt 600
agcggccagg gaccaggtca gcaggggcca ggaggtcagg gcccatacgg tccgggcgca 660
tccgcggcgg cggcagcggc aggtggctac ggtcccggaa gcggccaaca ggggccaggg 720
caacaaggac caggacaaca aggtcctggg ggccaaggac cgtatggacc aggagcatca 780
gctgcagccg cggcagctgg cggttacggt ccaggctacg gccagcaggg tccgggtcag 840
cagggaccgg gaggccaggg gccttatggc cctggcgctt ccgcagccag tgccgcttct 900
ggaggatacg ggccgggaag cggtcagcaa ggccctggcc aacaaggacc tggaggccaa 960
gggccctacg gcccaggagc ctcggcagcc gcagctgccg caggtgggta tgggccaggt 1020
agcgggcaac aagggccggg tcagcaagga ccggggcaac agggacctgg gcagcaagga 1080
cccgggggtc aaggcccgta cggacctggt gcgtctgcag ctgctgctgc ggctggtgga 1140
tatggtccgg gatcggggca gcagggtccc ggtcagcagg gccctggtca gcaagggcca 1200
ggccaacagg gacccggaca acaaggcccg ggtcaacagg gtcctggaca gcaggggccg 1260
ggccaacaag gccctgggca acagggtccg gggggacagg gggcctatgg gcctggcgca 1320
tctgccgccg ctggcgcagc cggtgggtac gggcctgggt caggtcaaca ggggcctggt 1380
caacaaggcc ccgggcaaca gggccccggc cagcaaggtc cagggcagca gggcccggga 1440
cagcaagggc ctggacaaca ggggcccgga cagcagggac cttacgggcc cggtgcgagc 1500
gcagcggccg ccgccgcagg gggatatggc cccggatcgg gccagcaggg accaggccag 1560
caaggacctg gccaacaggg cccggggggt caggggccgt atggtcccgg cgctgcaagt 1620
gctgcagtgt ccgtttctag agcacgagcc ggttcgggac aacaagggcc tggccagcag 1680
ggcccaggtc aacaagggcc aggacagcag ggtccttatg ggcccggcgc aagcgcagca 1740
gctgcggccg ctggtggcta tggtcctggc tccggtcaac agggcccttc gcaacaaggt 1800
cccgggcagc aaggtcctgg tggccagggt ccctacgggc cgggggcgag tgcggcagca 1860
gccgctgcag gcggttatgg tccaggaagc ggacagcaag gtccgggagg tcaaggtccg 1920
tatggcccag gctctagcgc ggctgccgct gccgcgggtg gcaacggacc agggagcgga 1980
caacagggcg cgggacaaca gggtccagga cagcaaggcc caggggcgtc ggcggctgca 2040
gcggcggccg gaggctatgg acccggctca ggacaacagg gaccgggtca acaaggaccc 2100
ggtggccaag gcccctatgg cccgggcgcc agcgcggccg cagccgccgc gggcgggtac 2160
ggccccggta gcggccaggg accaggtcag caggggccag gaggtcaggg cccatacggt 2220
ccgggcgcat ccgcggcggc ggcagcggca ggtggctacg gtcccggaag cggccaacag 2280
gggccagggc aacaaggacc aggacaacaa ggtcctgggg gccaaggacc gtatggacca 2340
ggagcatcag ctgcagccgc ggcagctggc ggttacggtc caggctacgg ccagcagggt 2400
ccgggtcagc agggaccggg aggccagggg ccttatggcc ctggcgcttc cgcagccagt 2460
gccgcttctg gaggatacgg gccgggaagc ggtcagcaag gccctggcca acaaggacct 2520
ggaggccaag ggccctacgg cccaggagcc tcggcagccg cagctgccgc aggtgggtat 2580
gggccaggta gcgggcaaca agggccgggt cagcaaggac cggggcaaca gggacctggg 2640
cagcaaggac ccgggggtca aggcccgtac ggacctggtg cgtctgcagc tgctgctgcg 2700
gctggtggat atggtccggg atcggggcag cagggtcccg gtcagcaggg ccctggtcag 2760
caagggccag gccaacaggg acccggacaa caaggcccgg gtcaacaggg tcctggacag 2820
caggggccgg gccaacaagg ccctgggcaa cagggtccgg ggggacaggg ggcctatggg 2880
cctggcgcat ctgccgccgc tggcgcagcc ggtgggtacg ggcctgggtc aggtcaacag 2940
gggcctggtc aacaaggccc cgggcaacag ggccccggcc agcaaggtcc agggcagcag 3000
ggcccgggac agcaagggcc tggacaacag gggcccggac agcagggacc ttacgggccc 3060
ggtgcgagcg cagcggccgc cgccgcaggg ggatatggcc ccggatcggg ccagcaggga 3120
ccaggccagc aaggacctgg ccaacagggc ccggggggtc aggggccgta tggtcccggc 3180
gctgcaagtg ctgcagtgtc cgttggaggt tacggccctc agtcttcgtc tgttccggtg 3240
gcgtccgcag ttgcgagtag actgtcttca cctgctgctt catcgcgagt atcgagcgct 3300
gtttcgtctc ttgtctcgtc gggtcccacg aaacatgccg ccctttcaaa tacgatttca 3360
tctgtagtgt cccaagttag tgcaagtaac ccggggttat ccggatgcga cgttctcgtt 3420
caggcactcc tagaagtagt atccgcgttg gtgagcatct tataa 3465
<210> 8
<211> 20
<212> DNA
<213> Artificial
<220>
<223> T7 promoter primer
<400> 8
taatacgact cactataggg 20
<210> 9
<211> 27
<212> DNA
<213> Artificial
<220>
<223> Rep Xba I primer
<400> 9
tctagaaacg gacactgcag cacttgc 27
<210> 10
<211> 28
<212> DNA
<213> Artificial
<220>
<223> Xba I Rep primer
<400> 10
tctagagcac gagccggttc gggacaac 28
<210> 11
<211> 19
<212> DNA
<213> Artificial
<220>
<223> T7 terminator primer
<400> 11
gctagttatt gctcagcgg 19
<210> 12
<211> 5
<212> PRT
<213> Artificial
<220>
<223> REP6-U1
<220>
<221> UNSURE
<222> (1)..(5)
<223> Xaa at position 4 or 5 is any one of amino acids, in particular,
it preferably is Ala, Ser, Tyr, Gln, Val, Leu or Ile, it more
preferably is Ala, Ser, Tyr, Gln or Val.
<400> 12
Gly Pro Gly Xaa Xaa
1 5
<210> 13
<211> 5
<212> PRT
<213> Artificial
<220>
<223> REP6-U2
<220>
<221> UNSURE
<222> (1)..(5)
<223> Xaa at position 5 is any one of amino acids, in particular, it
preferably is Ala, Ser, Tyr, Gln, Val, Leu or Ile, it more
preferably is Ala, Ser, Tyr, Gln or Val.
<400> 13
Gly Pro Gly Gly Xaa
1 5