KR20210044241A - 에피머라제 효소 및 그의 용도 - Google Patents

에피머라제 효소 및 그의 용도 Download PDF

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KR20210044241A
KR20210044241A KR1020217006658A KR20217006658A KR20210044241A KR 20210044241 A KR20210044241 A KR 20210044241A KR 1020217006658 A KR1020217006658 A KR 1020217006658A KR 20217006658 A KR20217006658 A KR 20217006658A KR 20210044241 A KR20210044241 A KR 20210044241A
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윌리엄 슈뢰더
파드메쉬 벤키타수브라마니안
스리다르 고빈다라잔
마크 웰치
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Abstract

본 개시내용은 프룩토스로부터 알룰로스의 상업적 규모의 생성에 유용한 에피머라제 효소를 제공한다. 개시된 효소("에피머라제 변이체")는, 모 효소와 비교하여 약 1.5배 내지 2배의 개선된 촉매 활성을 갖도록 조작된 부르크홀데리아 멀티보란스(Burkholderia multivorans) CGD1 자일로스 이성질화효소의 변이체이다.

Description

에피머라제 효소 및 그의 용도
관련 출원의 상호 참조
본 출원은 그 전체가 본 명세서에 참고로 포함된 2018년 8월 8일 제출된 제62/716,204호에 대한 우선권을 주장한다. 본 개시내용에 인용된 각각의 참고문헌, 특허, 및 공개 특허 출원은 본 명세서에 그 전체가 참고로 포함된다.
본 출원은 파일명이 "00820000058sequencelisting"인, 2019년 8월 5일에 생성된 178 kb의 텍스트 파일을 참고문헌으로 포함하며, 이는 본 출원의 서열목록이다.
기술분야
본 개시내용은 대체로 알룰로스의 생성에 관한 것이다.
Izumori et al., "Ketose 3-epimerase produced by Arthrobacter globiformis, method for converting ketose using the epimerase, and method for manufacture of ketose using the epimerase," WO 2014109254. Woodyer & Armentrout, "Recombinant ketose 3-epimerases for conversion of fructose to psicose," WO 2014049373
Baba et al., "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection," Molecular Systems Biology (2006) doi: l0/l038/msb4l00050, 11 pages Chu et al, "Expression, purification and activity of D-tagatose 3-epimerase from Clostridium bolteae," Shipin Gongye Keji 33, 198-201, 2012 Kim et al., "Characterization of an Agrobacterium tumefaciens D-psicose 3-epimerase that converts D-fructose to D-psicose," Applied and Environmental Microbiology 72, 981-85, 2006 Li et al., "Overexpression of -psicose 3-epimerase from Clostridium cellulolyticum H10 in Bacillus subtilis and its prospect for -psicose production," Advance Journal of Food Science and Technology 5, 264-69, 2013 Mu et al., "A strategy for bioproduction of rare sugars: Izumoring," Zhongguo Shengwu Gongcheng Zazhi 27, 129-36, 2007 Mu et al., "Recent advances on applications and biotechnological production of D-psicose," Applied Microbiology and Biotechnology 94, 1461-67, 2012 Phan et al, "Development of a strong intracellular expression system for Bacillus subtilis by optimizing promoter elements," J Biotechnol. 157, 167-72, 2012 Wagner et al, "Practical Aspects of Integrated Operation of Biotransformation and SMB Separation for Fine Chemical Synthesis," Organic Process Research & Development 16, 323-30, 2012 Zhang et al., "Characterization of a Metal-Dependent D-Psicose 3-Epimerase from a Novel Strain, Desmospora sp. 8437," J. Agricultural and Food Chemistry 61, 11468-76, 2013 Zhang et al., "Characterization of D-tagatose-3-epimerase from Rhodobacter sphaeroides that converts d-fructose into D-psicose," Biotechnology Letters 31, 857-62, 2009 Zhou et al., "Immobilization of D-tagatose 3-epimerase," Shipin Gongye Keji 33, 197-200, 2012
본 개시내용은 프룩토스로부터 알룰로스의 상업적 규모의 생성에 유용한 에피머라제 효소를 제공한다. 개시된 효소("에피머라제 변이체")는, 모 효소와 비교하여 약 1.5배 내지 2배의 개선된 촉매 활성을 갖도록 조작된 부르크홀데리아 멀티보란스(Burkholderia multivorans) CGD1 자일로스 이성질화효소(서열번호 6)의 변이체이다. 아래 표 1은 첨부된 서열 목록에서 각각의 에피머라제 변이체의 아미노산 서열을 식별하고, 그 변이체를 인코딩하는 핵산 서열의 예를 제공한다. 각각의 에피머라제 변이체와 모 효소(서열번호 6) 간의 아미노산 차이는 표 2에 요약되어 있으며, 여기서 아미노산 번호는 서열번호 6의 아미노산 번호를 나타낸다.
[표 1]
아미노산 서열 및 에피머라제 변이체의 코딩 서열의 예
Figure pct00001
[표 2]
에피머라제 변이체들
Figure pct00002
핵산, 벡터 및 숙주 미생물
본 개시내용은 상술한 개시된 에피머라제 변이체를 인코딩하는 핵산을 제공한다. 서열목록은 이들 변이체를 인코딩하는 뉴클레오티드 서열의 예를 제공하지만, 에피머라제 변이체를 인코딩하는 임의의 뉴클레오티드 서열이 사용될 수 있다.
뉴클레오티드 서열은, 당업계에 잘 알려진 바와 같은 미생물의 다양한 종 또는 균주에서의 발현에 대해 최적화될 수 있다. 적합한 미생물은 바실러스 리체니포르미스(Bacillus licheniformis), 사카로미세스 세레비시에(Saccharomyces cerevisiae), 스키조사카로미세스 폼베(Schizosaccharomyces pombe), 슈도모나스 퓨티다(Pseudomonas putida), 피치아 종(Pichia sp.), 아스페르길루스 종 (Aspergillus sp.), 트리코더마 레세이(Trichoderma reesei), 코리네박테리움 글루타미쿰(Corynebacterium glutamicum), 대장균(E. coli), 및 바실러스 서브틸리스 (B. subtilis)를 포함하지만, 이에 제한되지는 않는다. 이들 생물에서 임의의 단백질을 발현하기 위한 프로모터 및 기타 필요한 조절 서열을 함유하는 벡터는 알려져 있으며, 당업자가 쉽게 이용가능하다.
상술한 에피머라제 변이체를 인코딩하는 핵산은, 코딩 서열이 바람직한 숙주 미생물에서의 발현을 위해 적합한 조절 서열에 작동 가능하게 연결된 벡터 내에 포함될 수 있다. 조절 서열은 적합한 mRNA 리보솜 결합 부위, 및 전사와 번역의 종결을 조절하기 위한 서열을 포함하고, 프로모터 또는 오퍼레이터와 같은 다른 요소를 포함할 수 있다. 일단 숙주 미생물 내로 형질전환되면, 벡터는 숙주 게놈에 독립적으로 복제 또는 작용할 수 있거나 게놈 그 자체 내로 통합될 수 있다. 사용된 벡터는 특별히 제한되지 않으며, 숙주 내에서 복제할 수 있는 한, 당업계에 알려진 임의의 벡터일 수 있다.
벡터는 적어도 하나의 선택가능한 마커, 예컨대 항생제 내성 유전자를 포함할 수 있다. 적합한 항생제는, 예를 들어 아미카신, 암피실린, 오그멘틴(아목실린 + 클라불론산), 세파졸린, 세폭시틴, 세프타지딤, 세프티오푸르, 세팔로틴, 클로람페니콜, 엔로플록사신, 플로르페니콜, 젠타마이신, 이미페넴, 카나마이신, 페니실린, 사라플록시신, 스펙티노마이신, 스트렙토마이신, 테트라사이클린, 티카르실린, 및 틸미코신을 포함한다.
벡터는 상술한 에피머라제 변이체 중 하나 이상을 생성하기 위해 미생물을 조작하는 데 사용될 수 있다. 미생물로 벡터를 전달하는 방법은 잘 알려져 있으며, 예를 들어 인산칼슘 형질감염, DEAE-덱스트란 매개 형질감염, 현미주입 (microinjection), 지질-매개 형질감염, 전기천공법, 접합(conjugation) 및 감염을 포함한다.
알룰로스의 생성 방법
본 명세서에 개시된 에피머라제 변이체는 당업계에 잘 알려진 방법을 이용하여 프룩토스로부터 알룰로스의 상업적 규모의 생성을 위해 사용될 수 있다. 예를 들어, US 2017/0101637, US 2017/0298400, US 2016/0304853, 및 US 2016/0281076 참조.
일부 구현예에서, 에피머라제 변이체 또는 에피머라제 변이체를 포함하는 미생물로부터의 추출물은 고체 기질(matrix)에 결합되고, 프룩토스를 포함하는 유입 용액은 기질 위를 통과하여 프룩토스의 적어도 일부를 알룰로스로 전환시키며, 이 알룰로스는 배출 스트림으로부터 회수될 수 있다. 선택적으로, 알룰로스는 배출 스트림 내 다른 성분들로부터 분리될 수 있고 농축될 수 있다.
효소 결합에 적합한 많은 고체 기질은 당업계에 잘 알려져 있으며, 알지네이트(예를 들어, 나트륨 알지네이트), AMBERLITETM 수지(예를 들어, XAD®2, XAD®4, XAD®8, XAD®16(Sigma Aldrich), SEPHADEX® 수지 또는 DUOLITETM 수지(예를 들어, A568; Dow Chemical); 및 Purolite CR8415 및 ECR 8314(Purolite)를 포함한다. 또한 WO 2016/160573 및 WO 2016/191267 참조. 지지체 상에 에피머라제를 고정시키는 방법은 당업자에게 잘 알려져 있다. 예를 들어, 미국 특허 8,375,106 참조.
다른 구현예에서, 에피머라제 변이체를 발현하는 미생물은, 미국 특허 8,735,106에 설명된 바와 같은 알지네이트 비즈 상에, 또는 클레이, 탄소, 규조토, 또는 폴리아크릴아미드와 같은 수성 겔 상에 침투 및 고정될 수 있다.
당업자는 첨부된 청구범위의 범주에 속하는 상술한 구현예들의 수많은 변형 및 치환이 있음을 이해할 것이다.
도 1은 야생형 에피머라제(204020, 서열번호 6) 및 에피머라제 변이체들 230338(서열번호 16), 277285(서열번호 90), 277286(서열번호 92), 277287(서열번호 94), 277288(서열번호 96), 277289(서열번호 98), 및 277290(서열번호 100)에 의해 전환된 프룩토스의 분율에 대한 아미노산 치환의 효과를 보여주는 그래프이다. 각각의 쌍의 왼쪽 막대, 200 mM; 각각의 쌍의 오른쪽 막대, 50%(중량/중량).
도 2는 야생형 에피머라제(204020, 서열번호 6) 및 에피머라제 변이체들 230338(서열번호 16), 277285(서열번호 90), 277286(서열번호 92), 277287(서열번호 94), 277288(서열번호 96), 277289(서열번호 98), 및 277290(서열번호 100)의 발현을 나타내는 그래프이다.
도 3은 야생형 에피머라제(204020, 서열번호 6) 및 에피머라제 변이체들 230338(서열번호 16), 277285(서열번호 90), 277286(서열번호 92), 277287(서열번호 94), 277288(서열번호 96), 277289(서열번호 98), 및 277290(서열번호 100)의 비 활성도를 나타내는 그래프이다. 각각의 쌍의 왼쪽 막대, 200 mM; 각각의 쌍의 오른쪽 막대, 50% DS.
실시예 1. 알룰로스 에피머라제의 상업적 생성을 위한 마커가 없는 바실러스 서브틸리스 균주의 생성
본 실시예는 알룰로스 에피머라제를 발현하는 마커가 없는 바실러스 서브틸리스 균주의 생성을 설명한다. 간략하게는, 제1 단계에서, 바실러스 서브틸리스 균주는 BMCGD1 에피머라제를 인코딩하고 항생제 내성 마커를 포함하는 카세트로 형질전환되었다. 이 카세트는 바실러스 염색체로 재조합되어, 큰 포자형성 유전자 클러스터 및 리신 생합성 유전자 lysA를 포함하는, 8 kb의 DNA를 녹아웃시켰다(knocked out). 제2 단계에서, 제2 카세트를 바실러스 서브틸리스 염색체로 재조합하여, lysA 유전자를 복원하고 항생제 내성을 인코딩하는 DNA를 제거하였다. 바실러스 서브틸리스의 형질전환 전에, Keio 균주 은행으로부터의 대장균 균주 39 A10을 사용하여 플라스미드 DNA를 계대배양하였다. 관련 표현형은 다르게는 대장균의 야생형 K-12 균주에서 DNA 메틸라제 HsdM의 결손이다.
상세하게는, 5120 bp의 카세트(서열번호 1; 아이오와주 코랄빌 소재 IDT의 합성 DNA)를 합성하고, 표준 암피실린 내성 pIDT 벡터 내로 클로닝하였다. 합성 조각은 바실러스 서브틸리스 염색체 상 lysA의 업스트림 700 bp, 항생제 마커 cat (651 bp), DNA-결합 단백질 lacI(1083 bp), 및 알룰로스 에피머라제(894 bp)를 인코딩하였으며, dacF에서 상동성인 700 bp를 포함하였다. 이 벡터를 대장균 균주 39 A10(Baba 등, 2006) 내로 형질전환시켰으며, 플라스미드 DNA를 제조하여 바실러스 서브틸리스 균주 1A751 및 1A976 내로 형질전환시켰다.
클로람페니콜로 보충된 LB 상에서 형질전환체를 선택하였다. pIDT에 대한 레플리콘(replicon)은 대장균에서 작용성이지만, 바실러스 서브틸리스와 같은 그람 양성 세균에서는 작용하지 않는다. 따라서 발생하는 콜로니는 염색체 내로의 통합 이벤트(event)를 나타내었다. 균주 1A751에서, 플레이트 상에서 콜로니 형태학을 이용하여 단일 및 이중 재조합 이벤트를 구분하였다. 이중 재조합 이벤트는 포자형성에 필요한 유전자를 녹아웃시키는 반면, 단일 재조합은 그렇지 않을 것이다. LB 플레이트 상에서 3일 후, 포자형성가능한 콜로니는 불투명한 갈색이었다; 포자형성-결핍 콜로니는 더욱 반투명하였다.
알룰로스 에피머라제 카세트를 갖는 바실러스 서브틸리스 균주 1A976은 히스티딘 및 리신에 대해 영양요구성이며 자일로스 유도에 대해 매우 높은 형질전환 효율을 달성할 수 있다. 1925 bp 합성 DNA(서열번호 2)는 프라이머(서열번호 3, 서열번호 4) 및 Taq 폴리머라제(Promega)에 의해 증폭되었다. 이 PCR 생성물은 에피머라제 카세트 내에서 빠짐으로써 결실된 lysA 유전자, 및 lacI에 대한 상동성인 500 bp를 인코딩하였다. 이 DNA의 성공적인 이중 재조합 이벤트는 리신에 대해 독립영양성이며 클로람페니콜에 대해 민감성인 콜로니를 생성할 것이며; 즉 전체 cat 유전자가 손실될 것이다.
형질전환체는 히스티딘으로 보충된 Davis 최소 배지 상에서 선택되었다. 발생한 콜로니는 PCR에 의해 및 클로람페니콜이 있는 LB 및 없는 LB 상에서 스트리킹함으로써 특성화되었다. 도입된 DNA를 증폭시키고 클로람페니콜 민감성인 균주를 추가로 특성화하여, 염색체 DNA를 추출하였다.
클로람페니콜 내성 알룰로스를 함유하는 균주 1A751을 이 염색체 DNA를 이용하여 형질전환시키고 히스티딘으로 보충된 Davis 최소 배지 상에서 선택하였다. 형질전환체를 클로람페니콜이 있는 LB 및 클로람페니콜이 없는 LB 상에서 스트리킹하고 아래 설명되는 바와 같이 효소적으로 특성화하였다.
실시예 2. 에피머라제 단백질 수준의 분석
4 내지 12% 비스-트리스 뉴페이지(NUPAGE®) 겔(Invitrogen) 상에서 폴리아크릴아미드 겔 전기영동에 의해 조 가용성 용해물에서의 에피머라제 단백질 수준을 분석하였다. 단백질 수준을 단백질 정량 표준을 이용하여, 심플리블루 (SIMPLYBLUETM) 세이프 스테인(Invitrogen)으로 염색된 겔의 밀도측정에 의해 결정하였다.
실시예 3. 바실러스 서브틸리스 에서의 BMCGD1 에피머라제의 발현
BMCGD1 에피머라제 유전자를 포함하는 pHT254 작제물을 바실러스 서브틸리스 (서열번호 5)에서의 발현에 대해 코돈 최적화하고, 바실러스 서브틸리스 균주 DP1077을 형질전환하는 데 사용하였다. 균주 DP1077은 바실러스 유전 스톡 센터 (Bacillus Genetic Stock Center)의 균주 1A976(Em his nprE18 aprE3 eglS(DELTA)102 bglT/bglS(DELTA)EV lacA::PxylA-comK)의 포자형성-결함 (spoIIG::ZeoR) 유도체이다. 포자형성 결함에 더하여, 상기 균주는 각각 nprE 및 aprE 유전자에서의 돌연변이의 결과로서, 천연 프로테아제 및 서브틸리신을 분비하는 능력에 결함이 있다. 균주는 추가로, 컴피턴트(competent) 세포의 단순 생성을 위해 자일로스-유도성 프로모터의 제어 하에, 컴피턴스 인자 comK가 배치하는 발현 카세트를 보유한다.
형질전환체를 선택하여, 1% 글루코스 및 5 μg/ml 클로람페니콜이 보충된 Mn2+ 및 Co2+가 없는 맞춤형 Azure 배지(Teknova), 또는 2 g/L 합성 완전 아미노산 혼합물(MP Biomedicals), 1% 글루코스, 및 5 ㎍/ml 클로람페니콜이 보충된 Davis 최소 배지(HiMedia) 중 어느 하나에서 배양하였다. 배양물을 37℃에서 16시간 동안 성장시켰다. 이 배양물 40 μL를 사용하여 2 ml의 신선한 매질을 접종하고, 결과 배양물을 37℃에서 미드-로그(mid-log) 성장(600 nm에서의 약 0.7의 OD)까지 인큐베이션시켰다. 이어서 1 mM IPTG로 배양물을 유도하고, 37℃에서 4시간 동안 또는 24℃에서 20시간 동안 인큐베이션을 지속하였다.
세포를 원심분리에 의해 수확하고, 2회 동결 및 해동하고, 페리프렙스 (PERIPREPSTM) 완전 용해 프로토콜(Epicentre)을 사용하여 용해시켰다. 조 용해물의 원심분리 후 상청액 분획을 수집하여 용해물 중의 가용성 단백질을 제조하였다.
실시예 2에서 상술한 바와 같이 조 및 가용성 용해물에서 에피머라제 단백질 수준을 폴리아크릴아미드 겔 전기영동에 의해 분석하였다.
실시예 4. 에피머라제 변이체의 생성
US 8,635,029에 기재된 방법에 기초하여, 아미노산 변이를 인코딩하는 뉴클레오티드 변화를 BMCGD1 최적화된 코딩 서열 내로 도입하고 에피머라제 변이체를 위한 코딩 서열을 생성하는 데 사용하였다. 이들 코딩 서열을 pHT254(MoBiTech, Inc.) 내로 조작하였다. 이 벡터는 바실러스 서브틸리스의 groESL 오페론에 선행하는 프로모터로부터 유래된 강력한 Pgrac100 프로모터로부터 관심 대상의 유전자를 발현시킨다. 이는 IPTG 및 강한 리보좀성 결합 부위에 의한 유도를 가능하게 하는 lac 오퍼레이터에 융합된 개선된 조절 요소를 함유한다. 뉴클레오티드는 UP 요소인, -35 및 -15 영역을 포함하는 groESL 프로모터의 보존된 영역에서 최적화되었다 (Phan 등, 2012). 각각의 코딩 서열은 BamHI 및 XmaI 제한 부위에서 발현 벡터 pHT254 내로 클로닝되었다.
이어서 상술한 바와 같이 벡터를 바실러스 서브틸리스 DB1077 균주 내로 형질전환시켰다. 형질전환체를 5 ㎍/mL 클로람페니콜을 함유하는 LB 한천 매질 상에서 선택하였다.
96개의 바실러스 서브틸리스 형질전환체를 선택하여 Davis 최소 배지에 두었다. Davis 최소 배지는 시약 등급의 물을 사용하여, 최종 부피 1 L, 10.6 g의 최소 브로쓰 Davis w/o 덱스트로스(HiMedia 카탈로그 번호 M390-500G)와 2 g의 합성 완전 아미노산 혼합물(MP Biomedicals 카탈로그 번호 4400-022)로 만들고, 121℃에서 15분 동안 오토클레이빙하였다. 사용 전에, 글루코스를 1%까지 첨가하고, 클로람페니콜을 5 ㎍/ml까지 첨가하였다.
실시예 5. 에피머라제 활성도에 대한 형질전환체의 스크리닝
D-프룩토스에서 D-알룰로스 에피머라제로의 스크리닝을 위하여, 형질전환체를 선택하여 2 g/L 합성 완전 아미노산(MP Biomedicals), 1% 글루코스, 및 5 ㎍/mL 클로람페니콜로 보충된 600 μL Davis 최소 배지(HiMedia)에 두었다. 세포를 37℃에서 미드-로그로 성장시키고, 이어서 IPTG로 24℃에서 20시간 동안 유도하였다. 세포를 원심분리에 의해 수확하고, 페리프렙스 용해 프로토콜(Epicentre)을 사용하여 최종 부피 75 μL로 용해시켰다. 가용성 단백질 발현은 4% 내지 12% 비스-트리스 (Bis-Tris) 뉴페이지 겔(Invitrogen) 상에서 폴리아크릴아미드 겔 전기영동에 의해 분석하고, 단백질 수준을 정량 표준에 대한 밀도 측정에 의해 결정하였다. 회수된 가용성 에피머라제는 1 내지 10 ㎍/mL의 배양물 범위였다.
바실러스 서브틸리스 유래 용해물의 에피머라제 활성도에 대하여, 반응물을 100 μL 부피로 분석하였다: 10% 부피/부피 정화된 용해물, 저(200 mM) 또는 고 (50% 중량/중량) 프룩토스, 및 5 mM MgCl2. 반응물을 70℃에서 2시간 동안 인큐베이션시키고, 10% 부피의 2% HCl을 첨가하고, 4℃까지 냉각시켜 중단시켰다. HPLC 분석 전에 10 kDa 분자량 컷오프(Pall)의 PES 멤브레인을 통해 반응물을 여과했다.
결과는 표 3에 나타낸다.
[표 3]
에피머라제 변이체의 발현 및 프룩토스에서 알룰로스로의 2 hr 전환 프로파일
Figure pct00003
실시예 6. 야생형 에피머라제와, 1, 2 또는 3개의 아미노산 치환을 갖는 에피머라제 변이체의 비교 분석
변이체 230338에서 1, 2, 또는 3개의 아미노산 치환을 갖는 에피머라제 변이체(D89K, S154T, 및 C167I)에 의해 알룰로스로 전환된 프룩토스의 분획을 두 가지 조건(10% 부피/부피 정화된 용해물, 저(200 mM) 또는 고(50% 중량/중량) 프룩토스, 50 mM MES, pH 5.0, 및 5 mM MgCl2) 하에서 시험하고 야생형 에피머라제와 비교하였다. 결과는 도 1에 나타나 있으며, 여기서 각각의 막대 쌍의 왼쪽 막대는, 200 mM 프룩토스로 시작하여 1시간 후 프룩토스의 알룰로스로의 전환 분율을 나타내고, 각각의 막대 쌍의 오른쪽 막대는 50% 중량/중량 프룩토스로 출발하여 2시간 후 프룩토스의 알룰로스로의 전환 분율을 나타낸다. 각각의 변이체에 존재하는 아미노산 치환은 막대 그래프 위의 격자에 나타나 있고, 변이체의 아미노산 서열은 서열번호 16(변이체 230338), 서열번호 90(변이체 277285), 서열번호 92(변이체 277286), 서열번호 94(변이체 277287), 서열번호 96(변이체 277288), 서열번호 98(변이체 27289), 및 서열번호 100(변이체 27290)로 제공된다. "204020"은 야생형 에피머라제(서열번호 6)이다. 에피머라제 발현(㎍/ml)은 도 2에 나타나 있다. 비 활성도(에피머라제의 ㎍/ml 당 전환된 프룩토스)는 도 3에 나타나 있다.
본 실시예는 효소가 평가된 조건 하에서 3중 변이체가 촉매적으로 가장 활성이 있음을 입증한다. 3개의 치환(D89K, S154T, 및 C167I)을 모두 갖는 변이체는 가장 높은 발현을 가졌으며, 200 mM 프룩토스 및 50%(중량/중량) 프룩토스 모두에서 가장 활성이 높은 효소였다.
SEQUENCE LISTING <110> Archer Daniels Midland Company <120> EPIMERASE ENZYMES AND THEIR USE <130> 008200.00058 <140> <141> <150> US 62/716,204 <151> 2018-08-08 <160> 100 <170> FastSEQ for Windows Version 4.0 <210> 1 <211> 5120 <212> DNA <213> Artificial Sequence <220> <223> cassette <400> 1 taagatgaaa aacgctgaca ttgactgggg acaggtcagt gaccagctag ataaagcgaa 60 agataagatt acgaaattca tagaatcaga tgaaggcaaa aacttcatcc agaaagtcat 120 tgatttcttc gtatctatct ggaatgcgat tgtatctata ttcaaataag aaaagcgccg 180 aaaaatcggc gtttctttta ttgctttaca cccgttttaa atggcagatc aagttttaca 240 atatcttcgt atgtttctcg cttcacgact aaatgagcct caccgttttc gacaaataca 300 acggcgggtc tcggaatacg gttataattg ttggccatgc tgtatccata agcgcctgta 360 caaaaaacgg caagaagatc gccttctttt acttccggca ggtcaatatc ccaaatcagc 420 atatctccgc tttcacagca ctttccggca attgataccg ttttgtcatg cgcttctccg 480 atcctgttgg ctgccgcagc ttcatattta gcttggtaaa gcgcaggacg aatattgtcg 540 ttcatgcctc cgtctacagc cacatattgg cggacacccg gcacttcttt ttgagagcca 600 accgtataaa gagttgtgcc tgcgtctccc acgagagaac ggcccggttc gatccaaatt 660 tccggaatgt caaaaccgta acgggaagca ttttctttca gcatgcttaa gttattggta 720 tgactggttt taagcgcaaa aaaagttgct ttttcgtacc tattaatgta tcgttttaga 780 aaaccgactg taaaaagtac agtcggcatt atctcatatt ataaaagcca gtcattaggc 840 ctatctgaca attcctgaat agagttcata aacaatcctg catgataacc atcacaaaca 900 gaatgatgta cctgtaaaga tagcggtaaa tatattgaat tacctttatt aatgaatttt 960 cctgctgtaa taatgggtag aaggtaatta ctattattat tgatatttaa gttaaaccca 1020 gtaaatgaag tccatggaat aatagaaaga gaaaaagcat tttcaggtat aggtgttttg 1080 ggaaacaatt tccccgaacc attatatttc tctacatcag aaaggtataa atcataaaac 1140 tctttgaagt cattctttac aggagtccaa ataccagaga atgttttaga tacaccatca 1200 aaaattgtat aaagtggctc taacttatcc caataaccta actctccgtc gctattgtaa 1260 ccagttctaa aagctgtatt tgagtttatc acccttgtca ctaagaaaat aaatgcaggg 1320 taaaatttat atccttcttg ttttatgttt cggtataaaa cactaatatc aatttctgtg 1380 gttatactaa aagtcgtttg ttggttcaaa taatgattaa atatctcttt tctcttccaa 1440 ttgtctaaat caattttatt aaagttcatt tgatatgcct cctaaatttt tatctaaagt 1500 gaatttagga ggcttacttg tctgctttct tcattagaat caatcctttt ttaaaagtca 1560 atattactgt aacataaata tatattttaa aaatatccca ctttatccaa ttttcgtttg 1620 ttgaactaat gggtgcttta gttgaagaat aaagaccaca ttaaaaaatg tggtcttttg 1680 tgttttttta aaggatttga gcgtagcgaa aaatcctttt ctttcttatc ttgataataa 1740 gggtaactat tgccgatcgt ccattccgac agcatcgcca gtcactatgg cgtgctgcta 1800 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 1860 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 1920 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcaggcctta 1980 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 2040 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg 2100 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct 2160 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga 2220 tggtggttga cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg 2280 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca 2340 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg 2400 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt 2460 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta 2520 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca 2580 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat 2640 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat 2700 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg 2760 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt 2820 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg 2880 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg 2940 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt 3000 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga 3060 catcgtataa cgttactggt ttcatcaaaa tcgtctccct ccgtttgaat atttgattga 3120 tcgtaaccag atgaagcact ctttccacta tccctacagt gttatggctt gaacaatcac 3180 gaaacaataa ttggtacgta cgatctttca gccgactcaa acatcaaatc ttacaaatgt 3240 agtctttgaa agtattacat atgtaagatt taaatgcaac cgttttttcg gaaggaaatg 3300 atgacctcgt ttccaccgga attagcttgg taccaaagga ggtaaggatc actagaaaat 3360 tttttaaaaa atctcttgac attggaaggg agatatgtta ttataagaat tgcggaattg 3420 tgagcggata acaattccca tataaaggag gaaggatcca tgaataagtt aggtgtgcac 3480 gcattagttt gggaagcggg ttggagtcgt gatgagtgtg ctcgtgccat cgcacgcacc 3540 gcggaaacgg gatttgactt cattgaagtc ccagcgttag atccggcaag cattgatgca 3600 gagtttactc gcagagagtt agaacgccat gggttgggag taaccttttc gcttggactc 3660 gatgcgcaaa cagacattag ttctggtgat ccggaacgcg cagcgcgggg taaagctaaa 3720 ttagacgatg tcttacgtgt agcccgggat tgtggagcga ctcatgtctg tggaattctc 3780 tatagtgcat ttcaaaaaaa tgcagttcct acgacccgtg cgggtgtcgc catggccgcg 3840 gacattttag gacaggtagc tgatactgca gcacaatacg gtattacttt ggggttagaa 3900 gtcgttaacc gttatgagtc aaatgtattg aacaccgcgt cacagggtgt tgaactttgc 3960 gagcgtattg gtcgtcccaa cgtaaaagtg catttagata cgtatcacat gaatattgaa 4020 gagagcgata ttatgagcgc gatcagagat acgggagaca ggctagggta tttccatatt 4080 ggcgattctc atcgtggtta cctgggttcc ggaaacgtcg actttactgc cgtatttagg 4140 gcgctggtat tttctggata tacgggaccg attacatttg agtcattctc cagtagagtc 4200 gtaggacaac cattagaggg aattctggca atttggcgta atttatggga agattcccgc 4260 gatctagcta gtcacgcctt agcatatact cgtgtccagt taaaatctgc tcaagaagcc 4320 ctaaaacaag ccgagcgcag tcgcttaccg tgataacccg ggcagcccgc ctaatgagcg 4380 ggcttttttc acgtcacgcg tccatggaga tctaagcttt cactttttgc ttcagggttt 4440 cagccgtatg gtgatcgagt tcgcctgtta atcgaataca aagcacagat tctttgacat 4500 tcatgtcaat tccaaggctc atgctcattc ctccttgata tgatcggata atgagtgttt 4560 cgatttcgac ggaatgaatt ccttcaccgt gacaaaacta gtggtcattc ggcataatta 4620 cttaaatttt gtccagtctc ccatcgtccg ctttaagaat gtgataaacc cggctttctt 4680 catatcttct tttgcagcaa caggactttc agcgagtact tctccatcct ttttcagaac 4740 aagagtgcca agctcttggc ctttttgaat cggagcacta atattgtcct tcatcttgat 4800 ttcttttttc acatcgttca tatcctcgcc ttttttcgtc aatattgaaa tcggctcaga 4860 tgtagtgagt tcgataaatt tttgtttccc ttttttgacc tttacttttg ctactgtttg 4920 atttcgttta tataaaggat gcgtttcata ttggctaaag gcgaagtcaa gcatttttgt 4980 cacttgcgcg tttctttctt taggcgtgct cgctccgaat acaaccgcta tggcccgcat 5040 gtttcctttt ttagccgaag cagtcagaca atatttcgct tcgcctgtat agcctgtttt 5100 tacgccgtct acaccaggat 5120 <210> 2 <211> 1925 <212> DNA <213> Artificial Sequence <220> <223> cassette <400> 2 ttattgcttt acacccgttt taaatggcag atcaagtttt acaatatctt cgtatgtttc 60 tcgcttcacg actaaatgag cctcaccgtt ttcgacaaat acaacggcgg gtctcggaat 120 acggttataa ttgttggcca tgctgtatcc ataagcgcct gtacaaaaaa cggcaagaag 180 atcgccttct tttacttccg gcaggtcaat atcccaaatc agcatatctc cgctttcaca 240 gcactttccg gcaattgata ccgttttgtc atgcgcttct ccgatcctgt tggctgccgc 300 agcttcatat ttagcttggt aaagcgcagg acgaatattg tcgttcatgc ctccgtctac 360 agccacatat tggcggacac ccggcacttc tttttgagag ccaaccgtat aaagagttgt 420 gcctgcgtct cccacgagag aacggcccgg ttcgatccaa atttccggaa tgtcaaaacc 480 gtaacgggaa gcattttctt tcacagcttc gataattttt tcaacgtatt cagtggcatg 540 aagcggttca tcatcttccg tataacgaat gccgaaacct cctccaagat tcagcacctt 600 ggatacaaat gaatatgaat ctctccattc gtctagtttt ttgaagattt tttccgctgc 660 taacacaaaa ccggccgtat caaagatttg cgagccgata tggcaatgga cacccagcag 720 ctgaatgtgt tccgattgta atacttgttc aatggcccgt tcagtttgtc cgttatgaag 780 atcgaaacca aactttgaat cttcctggcc cgttgtaatg tagtcatgcg tatgcgcttc 840 tactccgggc gtgatccgaa gaagaacatc gatggagtga cccgtttctt tacataggtc 900 ttcaagaagc gcgatttcat agaaattatc caccacaatg cagccgatgc ggtgctcaag 960 cgccatccgc agttcttccc tgctcttatt gtttccatga aagtggatgc gttctgccgg 1020 aaagcctgct gcaacagccg tatatagctc tcctccggat acgacatcta aagaaagtcc 1080 ctcttcctca gcgagctgaa tcattgcgac tgatgagaat gctttgctcg catatgccac 1140 ctgtgctttc agccctgcag aaataaacgc ctgcttaaag cttttagcac gctcacgtat 1200 taaagccaca tcatatacgt aaagaggtgt accatatttc tccgctaaat agagagcatc 1260 cacacctccg atttctaaat gaccatgttg attttgtctg cttgtgccgt gtaagaacaa 1320 tgtcattccc tctttctccg cttatggatg aaaggacagt tttatggact gtcccttcat 1380 aattaagaca tacattatca tactcttcag tcagtttcaa tgcgatcact gcccgctttc 1440 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 1500 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg 1560 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc 1620 cagcaggcga aaatcctgtt tgatggtggt tgacggcggg atataacatg agctgtcttc 1680 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat 1740 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat 1800 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc 1860 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg 1920 cagac 1925 <210> 3 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 ttattgcttt acacccgttt taaatgg 27 <210> 4 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 tctcactcgc aatcaaattc agccg 25 <210> 5 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> codon optimized sequence for xylose isomerase [Burkholderia multivorans CGD1] <400> 5 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attagacgat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgagt caaatgtatt gaacaccgcg 480 tcacagggtg ttgaactttg cgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 6 <211> 297 <212> PRT <213> Burkholderia multivorans <400> 6 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 7 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 230323 <400> 7 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaaat tccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcggcag gtaaagctaa attagacgat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaactttg cgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 8 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 230323 <400> 8 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Ile Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp His Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser Asn Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 9 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 230329 <400> 9 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaaat tccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attagacgat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 10 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 230329 <400> 10 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Ala Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser His Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Ala Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 11 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 230333 <400> 11 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcggcag gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgagt caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 12 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 230333 <400> 12 Met Asn Lys Phe Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Leu His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Pro Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 13 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 230335 <400> 13 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcggcag gtaaagctaa attagacgat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 14 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 230335 <400> 14 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Val Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Glu Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Leu Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 15 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 230338 <400> 15 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 16 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 230338 <400> 16 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Gly Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Leu Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Ile Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 17 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 256407 <400> 17 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca ctgcaaaaat atgcagttcc tacgaccgca 360 gatggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 18 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 256407 <400> 18 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Lys Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Leu Gln 100 105 110 Lys Tyr Ala Val Pro Thr Thr Ala Asp Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Thr Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Ile Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 19 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 230339 <400> 19 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaactttg cgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattgttttt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 20 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 230339 <400> 20 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Ser Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Glu Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Thr Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 21 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 248843 <400> 21 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca ctgcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattccgc agatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 22 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 248843 <400> 22 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Phe His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Ile Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Ile 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 23 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 248867 <400> 23 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta agacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggatca 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 24 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 248867 <400> 24 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Glu Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Val 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Val Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 25 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 248891 <400> 25 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcta tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atcaaggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgcacag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 26 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 248891 <400> 26 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ala Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Asp Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Cys Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 27 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 248907 <400> 27 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcta tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta agacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 28 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 248907 <400> 28 Met Asn Lys Leu Gly Val His Ala Leu Leu Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Ile Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Arg Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 29 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 248914 <400> 29 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattatgt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 30 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 248914 <400> 30 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Tyr 20 25 30 Asp Phe Ile Glu Leu Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Glu Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 31 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 248925 <400> 31 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agattttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgaccgca 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggatca 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 32 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 248925 <400> 32 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Glu Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Met 225 230 235 240 Glu Ala Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 33 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 256382 <400> 33 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaata tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca ctgcaaaaaa atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta agacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 34 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 256382 <400> 34 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Tyr 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp His Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Leu Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 35 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 256406 <400> 35 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacgtta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 36 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 256406 <400> 36 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Pro Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Val His Ile Gly 195 200 205 Asp Ser Asn Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 37 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 255290 <400> 37 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 38 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 255290 <400> 38 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Thr Tyr Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Leu 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 39 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 257999 <400> 39 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 40 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 257999 <400> 40 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Gly Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Leu Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Ile 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 41 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261731 <400> 41 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 42 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261731 <400> 42 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp His Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Pro Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Pro Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 43 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261732 <400> 43 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 44 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261732 <400> 44 Met Asn Lys Leu Gly Val His Ala Leu Leu Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Pro Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Val His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 45 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261733 <400> 45 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 46 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261733 <400> 46 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ala Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Ile Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Pro Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 47 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261734 <400> 47 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 48 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261734 <400> 48 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 49 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261735 <400> 49 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 50 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261735 <400> 50 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Ser Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Glu Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Leu 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 51 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261736 <400> 51 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 52 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261736 <400> 52 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Pro Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val Leu Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Ile Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 53 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261737 <400> 53 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 54 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261737 <400> 54 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Ser Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Phe Glu Ser Asn Leu Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 55 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261738 <400> 55 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 56 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261738 <400> 56 Met Asn Lys Ile Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Glu Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Pro Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 57 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261739 <400> 57 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 58 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261739 <400> 58 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Gly Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Asp Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val Leu Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 59 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261740 <400> 59 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 60 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261740 <400> 60 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Ile Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Ser Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Leu Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 61 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261741 <400> 61 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 62 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261741 <400> 62 Met Asn Lys Phe Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Phe His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Arg Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 63 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261742 <400> 63 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 64 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261742 <400> 64 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Leu Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Gly Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Ala Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 65 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261743 <400> 65 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 66 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261743 <400> 66 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Pro Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Ile Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Trp Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 67 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261744 <400> 67 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 68 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261744 <400> 68 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Pro Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Leu Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Glu Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 69 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261745 <400> 69 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 70 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261745 <400> 70 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Pro Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser His Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Trp Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 71 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261746 <400> 71 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agattttact cgcagagagt tagaaaaata tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 72 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261746 <400> 72 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Gly Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Leu Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Arg Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 73 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261747 <400> 73 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccgagaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 74 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261747 <400> 74 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Leu Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Ala Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Leu Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 75 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261748 <400> 75 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaata tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcacg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 76 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261748 <400> 76 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Tyr Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Leu Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Val 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 77 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261749 <400> 77 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccgagaa gcattgatgc agagtttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcacg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 78 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261749 <400> 78 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Leu His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Ile Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Leu 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 79 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261750 <400> 79 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccgagaa gcattgatgc agattttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 80 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261750 <400> 80 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Val Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Val Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Gly Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 81 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261751 <400> 81 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agattttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgaatct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatttac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 82 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261751 <400> 82 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Thr Gly His Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Leu Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 83 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261752 <400> 83 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccgagaa gcattgatgc agattttact cgcagagagt tagaaaaaca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagatggccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 84 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261752 <400> 84 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Phe Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Leu Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Leu Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 85 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261753 <400> 85 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaata tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa tgcgacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgacccgt 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcacg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 86 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261753 <400> 86 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Leu Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Val Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Leu Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 87 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 261754 <400> 87 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaaaaata tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaat atgcagttcc tacgaccgca 360 agaggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacggcagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcacg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 88 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 261754 <400> 88 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Met 225 230 235 240 Glu Cys Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Val Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 89 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 277285 <400> 89 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgagt caaatgtatt gaacaccgcg 480 tcacagggtg ttgaactttg cgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 90 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 277285 <400> 90 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Lys Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 91 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 277286 <400> 91 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attagacgat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaactttg cgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 92 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 277286 <400> 92 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Thr Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 93 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 277287 <400> 93 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attagacgat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgagt caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 94 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 277287 <400> 94 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Ile Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 95 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 277288 <400> 95 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaactttg cgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 96 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 277288 <400> 96 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Lys Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Thr Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Cys Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 97 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 277289 <400> 97 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attagacgat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgaga caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 98 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 277289 <400> 98 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Asp Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Thr Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Ile Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295 <210> 99 <211> 897 <212> DNA <213> Artificial Sequence <220> <223> coding sequence for variant 277290 <400> 99 atgaataagt taggtgtgca cgcattagtt tgggaagcgg gttggagtcg tgatgagtgt 60 gctcgtgcca tcgcacgcac cgcggaaacg ggatttgact tcattgaagt cccagcgtta 120 gatccggcaa gcattgatgc agagtttact cgcagagagt tagaacgcca tgggttggga 180 gtaacctttt cgcttggact cgatgcgcaa acagacatta gttctggtga tccggaacgc 240 gcagcgcggg gtaaagctaa attaaaagat gtcttacgtg tagcccggga ttgtggagcg 300 actcatgtct gtggaattct ctatagtgca tttcaaaaaa atgcagttcc tacgacccgt 360 gcgggtgtcg ccatggccgc ggacatttta ggacaggtag ctgatactgc agcacaatac 420 ggtattactt tggggttaga agtcgttaac cgttatgagt caaatgtatt gaacaccgcg 480 tcacagggtg ttgaacttat tgagcgtatt ggtcgtccca acgtaaaagt gcatttagat 540 acgtatcaca tgaatattga agagagcgat attatgagcg cgatcagaga tacgggagac 600 aggctagggt atttccatat tggcgattct catcgtggtt acctgggttc cggaaacgtc 660 gactttactg ccgtatttag ggcgctggta ttttctggat atacgggacc gattacattt 720 gagtcattct ccagtagagt cgtaggacaa ccattagagg gaattctggc aatttggcgt 780 aatttatggg aagattcccg cgatctagct agtcacgcct tagcatatac tcgtgtccag 840 ttaaaatctg ctcaagaagc cctaaaacaa gccgagcgca gtcgcttacc gtgataa 897 <210> 100 <211> 297 <212> PRT <213> Artificial Sequence <220> <223> variant 277290 <400> 100 Met Asn Lys Leu Gly Val His Ala Leu Val Trp Glu Ala Gly Trp Ser 1 5 10 15 Arg Asp Glu Cys Ala Arg Ala Ile Ala Arg Thr Ala Glu Thr Gly Phe 20 25 30 Asp Phe Ile Glu Val Pro Ala Leu Asp Pro Ala Ser Ile Asp Ala Glu 35 40 45 Phe Thr Arg Arg Glu Leu Glu Arg His Gly Leu Gly Val Thr Phe Ser 50 55 60 Leu Gly Leu Asp Ala Gln Thr Asp Ile Ser Ser Gly Asp Pro Glu Arg 65 70 75 80 Ala Ala Arg Gly Lys Ala Lys Leu Lys Asp Val Leu Arg Val Ala Arg 85 90 95 Asp Cys Gly Ala Thr His Val Cys Gly Ile Leu Tyr Ser Ala Phe Gln 100 105 110 Lys Asn Ala Val Pro Thr Thr Arg Ala Gly Val Ala Met Ala Ala Asp 115 120 125 Ile Leu Gly Gln Val Ala Asp Thr Ala Ala Gln Tyr Gly Ile Thr Leu 130 135 140 Gly Leu Glu Val Val Asn Arg Tyr Glu Ser Asn Val Leu Asn Thr Ala 145 150 155 160 Ser Gln Gly Val Glu Leu Ile Glu Arg Ile Gly Arg Pro Asn Val Lys 165 170 175 Val His Leu Asp Thr Tyr His Met Asn Ile Glu Glu Ser Asp Ile Met 180 185 190 Ser Ala Ile Arg Asp Thr Gly Asp Arg Leu Gly Tyr Phe His Ile Gly 195 200 205 Asp Ser His Arg Gly Tyr Leu Gly Ser Gly Asn Val Asp Phe Thr Ala 210 215 220 Val Phe Arg Ala Leu Val Phe Ser Gly Tyr Thr Gly Pro Ile Thr Phe 225 230 235 240 Glu Ser Phe Ser Ser Arg Val Val Gly Gln Pro Leu Glu Gly Ile Leu 245 250 255 Ala Ile Trp Arg Asn Leu Trp Glu Asp Ser Arg Asp Leu Ala Ser His 260 265 270 Ala Leu Ala Tyr Thr Arg Val Gln Leu Lys Ser Ala Gln Glu Ala Leu 275 280 285 Lys Gln Ala Glu Arg Ser Arg Leu Pro 290 295

Claims (8)

  1. 단백질을 인코딩하는 핵산으로서, 여기서 단백질의 아미노산 서열은, 서열번호 16, 18, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 및 100으로 이루어진 군으로부터 선택되는 핵산.
  2. 제1항의 핵산을 포함하는 미생물.
  3. 제2항에 있어서, 미생물은 바실러스 리체니포르미스(Bacillus licheniformis), 사카로미세스 세레비시에(Saccharomyces cerevisiae), 스키조사카로미세스 폼베(Schizosaccharomyces pombe), 슈도모나스 퓨티다(Pseudomonas putida), 피치아 종(Pichia sp.), 아스페르길루스 종(Aspergillus sp.), 트리코더마 레세이(Trichoderma reesei), 코리네박테리움 글루타미쿰(Corynebacterium glutamicum), 대장균(E. coli), 및 바실러스 서브틸리스(B. subtilis)로 이루어진 군으로부터 선택되는, 미생물.
  4. 서열번호 16, 18, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 및 100으로 이루어진 군으로부터 선택되는 아미노산 서열을 포함하는 단백질.
  5. 제4항의 단백질을 포함하는 고체 기질(matrix).
  6. 제5항의 고체 기질을 포함하고, 고체 기질 위에 프룩토스를 포함하는 유입 용액을 수용하고 알룰로스를 포함하는 배출 용액의 배출을 허용하도록 구성된 컬럼.
  7. (a) 제2항의 미생물을 배양하는 단계; 및
    (b) 단백질을 회수하는 단계
    를 포함하는,
    서열번호 16, 18, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 및 100으로 이루어진 군으로부터 선택되는 아미노산 서열을 포함하는 단백질의 생성 방법.
  8. 프룩토스의 적어도 일부를 알룰로스로 전환시키기에 적합한 시간 동안 및 조건 하에서, 프룩토스를 포함하는 용액을 제4항의 단백질과 접촉시키는 단계를 포함하는 알룰로스의 생성 방법.
KR1020217006658A 2018-08-08 2019-08-07 에피머라제 효소 및 그의 용도 KR20210044241A (ko)

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Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113249287B (zh) * 2021-07-07 2021-09-24 中国科学院天津工业生物技术研究所 一株表达d-阿洛酮糖3-差向异构酶的枯草芽孢杆菌工程菌株及其应用
CN114277049A (zh) * 2021-12-31 2022-04-05 李宪臻 一种异源表达蔗糖异构酶的基因工程菌及应用

Family Cites Families (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8005620B2 (en) 2003-08-01 2011-08-23 Dna Twopointo Inc. Systems and methods for biopolymer engineering
KR20110035805A (ko) 2009-09-30 2011-04-06 씨제이제일제당 (주) 사이코스-에피머화 효소의 고정화 및 이를 이용한 사이코스의 제조방법
US9128961B2 (en) 2010-10-28 2015-09-08 Google Inc. Loading a mobile computing device with media files
GB2508586B (en) 2012-09-27 2020-09-02 Tate & Lyle Ingredients Americas Llc A protein
CN108315316B (zh) 2013-01-08 2021-07-20 松谷化学工业株式会社 球形节杆菌生产的酮糖3-差向异构酶
JP6774875B2 (ja) 2013-09-03 2020-10-28 ロケット フレールＲｏｑｕｅｔｔｅ Ｆｒｅｒｅｓ Ｄ−プシコース ３−エピメラーゼの改良された変異体およびその使用
KR101539097B1 (ko) 2013-12-26 2015-07-23 주식회사 삼양제넥스 사이코스 에피머화 효소를 암호화 하는 폴리뉴클레오타이드, 및 이를 이용하는 사이코스 생산방법
CN103710330B (zh) * 2014-01-03 2015-04-22 江南大学 一种高催化活性的d-阿洛酮糖 3-差向异构酶的突变体酶及其应用
KR101473918B1 (ko) 2014-05-28 2014-12-17 대상 주식회사 사이코스 에피머화 효소, 이의 제조방법 및 이를 이용한 사이코스의 제조방법
KR101577147B1 (ko) * 2014-10-01 2015-12-11 경상대학교산학협력단 사이코스의 생산 방법
WO2016160573A1 (en) 2015-03-27 2016-10-06 Archer Daniels Midland Co. Fructose to allulose conversion
ES2911890T3 (es) * 2015-05-22 2022-05-23 Archer Daniels Midland Co Uso de enzimas epimerasas para la conversión de fructosa en alulosa

Non-Patent Citations (13)

* Cited by examiner, † Cited by third party
Title
Baba et al., "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection," Molecular Systems Biology (2006) doi: l0/l038/msb4l00050, 11 pages
Chu et al, "Expression, purification and activity of D-tagatose 3-epimerase from Clostridium bolteae," Shipin Gongye Keji 33, 198-201, 2012
Izumori et al., "Ketose 3-epimerase produced by Arthrobacter globiformis, method for converting ketose using the epimerase, and method for manufacture of ketose using the epimerase," WO 2014109254.
Kim et al., "Characterization of an Agrobacterium tumefaciens D-psicose 3-epimerase that converts D-fructose to D-psicose," Applied and Environmental Microbiology 72, 981-85, 2006
Li et al., "Overexpression of -psicose 3-epimerase from Clostridium cellulolyticum H10 in Bacillus subtilis and its prospect for -psicose production," Advance Journal of Food Science and Technology 5, 264-69, 2013
Mu et al., "A strategy for bioproduction of rare sugars: Izumoring," Zhongguo Shengwu Gongcheng Zazhi 27, 129-36, 2007
Mu et al., "Recent advances on applications and biotechnological production of D-psicose," Applied Microbiology and Biotechnology 94, 1461-67, 2012
Phan et al, "Development of a strong intracellular expression system for Bacillus subtilis by optimizing promoter elements," J Biotechnol. 157, 167-72, 2012
Wagner et al, "Practical Aspects of Integrated Operation of Biotransformation and SMB Separation for Fine Chemical Synthesis," Organic Process Research & Development 16, 323-30, 2012
Woodyer & Armentrout, "Recombinant ketose 3-epimerases for conversion of fructose to psicose," WO 2014049373
Zhang et al., "Characterization of a Metal-Dependent D-Psicose 3-Epimerase from a Novel Strain, Desmospora sp. 8437," J. Agricultural and Food Chemistry 61, 11468-76, 2013
Zhang et al., "Characterization of D-tagatose-3-epimerase from Rhodobacter sphaeroides that converts d-fructose into D-psicose," Biotechnology Letters 31, 857-62, 2009
Zhou et al., "Immobilization of D-tagatose 3-epimerase," Shipin Gongye Keji 33, 197-200, 2012

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