KR102301138B1 - 옥신토모듈린 생산용 융합태그 - Google Patents

옥신토모듈린 생산용 융합태그 Download PDF

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KR102301138B1
KR102301138B1 KR1020200168669A KR20200168669A KR102301138B1 KR 102301138 B1 KR102301138 B1 KR 102301138B1 KR 1020200168669 A KR1020200168669 A KR 1020200168669A KR 20200168669 A KR20200168669 A KR 20200168669A KR 102301138 B1 KR102301138 B1 KR 102301138B1
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김성건
김윤정
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주식회사 펩진
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    • C—CHEMISTRY; METALLURGY
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    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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    • C07—ORGANIC CHEMISTRY
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    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand

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Abstract

본 발명의 일실시예는, 융합태그, 상기 융합태그와 옥시토모듈린을 포함하는 융합 폴리펩타이드에 관한 것이다. 본 발명에 따른 신규한 융합태그를 사용하면, 종래 융합태그 대비 옥시토모듈린의 수율을 향상시킬 수 있다. 특히, 상대적으로 분자량이 작으며 분해되기 쉬운 아미노 말단을 갖는 옥시토모듈린을 유전자 재조합 기술을 이용하여 제조하는 경우에 유용하다. 또한, 본 발명의 일실시예에 따른 융합태그를 포함하는 융합 폴리펩타이드는 숙주세포 내에서 내포체 형태로 발현이 유도됨으로써 숙주세포의 단백질 분해효소 등으로부터 보호되어 안정적으로 옥시토모듈린을 생산할 수 있는 장점이 있다. 따라서, 종래의 융합태그를 사용한 경우보다 안정성 및 수율이 향상된 옥시토모듈린 생산 방법을 제공할 수 있다.

Description

옥신토모듈린 생산용 융합태그{FUSION TAG FOR PREPARING OXYNTOMODULIN}
본 발명은 신규한 융합태그로서, 상기 융합태그와 옥시토모듈린을 포함하는 융합 폴리펩타이드에 관한 것이다.
최근 유전공학 및 생명공학 기술의 발달로 대장균, 효모, 동식물세포 등을 이용하여 유용한 외래 단백질이 많이 생산되고, 이들 단백질이 의약품 등으로 널리 이용되고 있다. 구체적으로, 면역 조절제, 효소 저해제 및 호르몬 같은 의약 및 연구용 단백질이나 반응 첨가 효소와 같은 산업용 단백질에 대한 생산 공정 기술 개발 및 산업화가 추진되고 있다.
이중에서도 유전자 재조합 기술은 여러 목적 단백질들의 핵산을 발현벡터에 클로닝하여 재조합 발현벡터를 얻고, 이를 적당한 숙주세포에 형질전환시켜 배양함으로써 목적 단백질 또는 목적 펩타이드를 생산하는 방법이다. 다만, 숙주세포에 내재된 분해 효소(예를 들어, 프로테아제(protease) 또는 펩티다제(peptidase))에 의해 목적 단백질 또는 목적 펩타이드의 전체 또는 일부가 분해되어 수율이 낮아지거나, 융합태그로 사용하는 펩타이드의 크기가 제조하고자 하는 목적 단백질 또는 목적 펩타이드의 크기에 비해 너무 커서 수율이 현저히 떨어지는 문제가 있다.
따라서, 유전자 재조합 기술을 이용하여 목적 단백질 또는 목적 펩타이드를 대량 생산하고자 하는 경우, 목적 단백질 또는 목적 펩타이드를 안정적으로 발현시키면서도 생산 수율을 향상시킬 수 있는 융합태그를 개발하는 것이 중요하다.
본 발명이 이루고자 하는 기술적 과제는 옥시토모듈린을 생산하기 위한 아미노산 서열로 이루어진 융합태그를 제공하는 것이다.
본 발명의 다른 목적은 상기 융합태그와 옥시토모듈린을 포함하는 융합 폴리펩타이드를 제공하는 것이다.
본 발명의 또 다른 목적은 상기 융합 폴리펩타이드를 코딩하는 뉴클레오타이드, 상기 뉴클레오타이드를 포함하는 발현벡터 및 상기 발현벡터를 포함하는 숙주세포를 제공하는 것이다.
본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 기술적 과제로 제한되지 않으며, 언급되지 않은 또 다른 기술적 과제들은 아래의 기재로부터 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자에게 명확하게 이해될 수 있을 것이다.
상기 기술적 과제를 달성하기 위하여, 본 발명의 일측면에 따른 옥시토모듈린 생산용 융합태그는 서열번호1, 서열번호2, 서열번호3, 서열번호4, 서열번호 5 및 서열번호6의 아미노산 서열을 포함할 수 있다.
본 발명의 실시예에 있어서, 상기 융합태그는 옥시토모듈린의 N말단에 위치할 수 있다.
본 발명의 실시예에 있어서, 상기 융합태그는 옥시토모듈린과 함께 발현되어 봉입체(inclusion body)를 형성할 수 있다.
본 발명의 실시예에 있어서, 상기 융합태그는 상기 융합태그와 상기 옥시토모듈린을 연결시켜주는 링커를 더 포함할 수 있다.
본 발명의 실시예에 있어서, 상기 링커는 친화성 태그를 포함할 수 있다.
본 발명의 실시예에 있어서, 상기 링커는 프로테아제 인식 서열을 포함할 수 있다.
본 발명의 실시예에 있어서, 상기 프로테아제 인식 서열은 상기 링커의 C말단에 위치할 수 있다.
본 발명의 실시예에 있어서, 상기 프로테아제 인식 서열은 담배 식각 바이러스 프로테아제 인식 서열, 엔테로키나아제 인식 서열, 유비퀴틴 카르복시 말단 가수분해효소 인식 서열, 인자Xa인식 서열, 퓨린 인식 서열 및 이들의 조합으로 이루어진 군으로부터 선택될 수 있다.
본 발명의 실시예에 있어서, 상기 프로테아제 인식 서열은 서열번호 9의 아미노산 서열을 포함할 수 있다.
본 발명의 다른 측면에 따른 뉴클레오타이드는 본 발명에 따른 융합태그; 및 옥시토모듈린을 포함할 수 있다.
본 발명의 또다른 측면에 따른 발현벡터는 본 발명에 따른 뉴클레오타이드를 포함할 수 있다.
본 발명의 실시예에 있어서, 상기 발현벡터는 서열번호 171의 염기서열을 포함할 수 있다.
본 발명의 또다른 측면에 따른 숙주세포는 본 발명에 따른 발현벡터를 포함할 수 있다.
본 발명의 실시예에 따른 신규한 융합태그를 사용하면, 종래 융합태그 대비 옥시토모듈린의 수율을 향상시킬 수 있다. 특히, 상대적으로 분자량이 작으며 분해되기 쉬운 아미노 말단을 갖는 옥시토모듈린을 유전자 재조합 기술을 이용하여 제조하는 경우에 유용하다. 또한, 본 발명의 실시예에 따른 융합태그를 포함하는 융합 폴리펩타이드는 숙주세포 내에서 내포체 형태로 발현이 유도됨으로써 숙주세포의 단백질 분해효소 등으로부터 보호되어 안정적으로 옥시토모듈린을 생산할 수 있는 장점이 있다. 따라서, 종래의 융합태그를 사용한 경우보다 안정성 및 수율이 향상된 옥시토모듈린 생산 방법을 제공할 수 있다.
본 발명의 효과는 상기한 효과로 한정되는 것은 아니며, 본 발명의 상세한 설명 또는 특허청구범위에 기재된 발명의 구성으로부터 추론 가능한 모든 효과를 포함하는 것으로 이해되어야 한다.
도 1은 pSK03 플라스미드의 모식도이다(lacIq: lac 억제자, Ptrc: trc 프로모터, RBS: 리보솜 결합부위, Kan R: 카나마이신 내성 유전자, pBR322 ori: pBR322 복제원점).
도2은 재조합 대장균에서 발현된 H6TEV-OXM, IMCD1-H6TEV-OXM, SUMO-H6TEV-OXM의 총세포 분획을 SDS-PAGE로 분석한 결과이다 (레인 M: 마커 단백질, 레인 1: H6TEV-OXM (균주 번호 PG095), 레인 2: IMC-D1-H6TEV-OXM (균주 번호 PG101), 레인 3: SUMO-H6TEV-OXM (균주 번호 PG096)).
도3은 재조합 대장균에서 발현된 IMCD1-H6TEV-OXM의 가용성을 SDS-PAGE로 분석한 결과이다 (레인 M: 마커 단백질, 레인 T: 총 세포 단백질, 레인 S: 가용성 세포 단백질, 레인 I: 불용성 세포 단백질).
도4는 재조합 대장균에서 발현된 OXM 융합 폴리펩타이드들의 총 세포 분획을 SDS-PAGE로 분석한 결과이다 (레인 M: 마커 단백질, 레인 1: H6TEV-OXM (균주 번호 PG095), 레인 2: IMCB1-H6TEV-OXM (균주 번호 PG097), 레인 3: IMCD1-H6TEV-OXM (균주 번호 PG101)).
도5은 크로마토그래피를 통해 정제한 IMC-B1-H6TEV-OXM 융합 폴리펩타이드를 SDS-PAGE로 분석한 결과이다. (레인 M: 마커 단백질, 레인 S: 정제 전 시료, 레인 FT: 통과액 분획, 레인 E: 용출액 분획). 화살표는 IMC-B1-H6TEV-OXM 융합 폴리펩타이드를 나타낸다.
도6은 TEV 프로테아제를 이용하여 정제된 IMCB1-H6TEV-OXM 를 절단한 후 분획을 SDS-PAGE로 분석한 결과이다 (레인 M: 마커 단백질, 레인 C: TEV 프로테아제를 처리하지 않은 시료, 레인 T: TEV 프로테아제를 처리한 시료).
도7는 역상 크로마토그래피를 이용하여 IMC-B1-H6TEV-OXM 로부터 절단된 IMC-B1-H6TEV와 OXM을 분리한 분획들을 SDS-PAGE로 분석한 결과이다 (레인 M: 마커 단백질, 레인 S: 정제 전 시료, 레인 FT: 통과액 분획, 레인 E: 용출액 분획).
도8은 본 발명에 따라 정제된 OXM의 분자량을 측정한 결과를 나타낸 것이다.
이하에서는 첨부한 도면을 참조하여 본 발명을 설명하기로 한다. 그러나 본 발명은 여러 가지 상이한 형태로 구현될 수 있으며, 따라서 여기에서 설명하는 실시예로 한정되는 것은 아니다. 그리고 도면에서 본 발명을 명확하게 설명하기 위해서 설명과 관계없는 부분은 생략하였으며, 명세서 전체를 통하여 유사한 부분에 대해서는 유사한 도면 부호를 붙였다.
명세서 전체에서, 어떤 부분이 다른 부분과 "연결(접속, 접촉, 결합)"되어 있다고 할 때, 이는 "직접적으로 연결"되어 있는 경우 뿐 아니라, 그 중간에 다른 부재를 사이에 두고 "간접적으로 연결"되어 있는 경우도 포함한다. 또한 어떤 부분이 어떤 구성요소를 "포함"한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성요소를 제외하는 것이 아니라 다른 구성요소를 더 구비할 수 있다는 것을 의미한다.
본 명세서에서 사용한 용어는 단지 특정한 실시예를 설명하기 위해 사용된 것으로, 본 발명을 한정하려는 의도가 아니다. 단수의 표현은 문맥상 명백하게 다르게 뜻하지 않는 한, 복수의 표현을 포함한다. 본 명세서에서, "포함하다" 또는 "가지다" 등의 용어는 명세서상에 기재된 특징, 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것이 존재함을 지정하려는 것이지, 하나 또는 그 이상의 다른 특징들이나 숫자, 단계, 동작, 구성요소, 부품 또는 이들을 조합한 것들의 존재 또는 부가 가능성을 미리 배제하지 않는 것으로 이해되어야 한다.
이하 첨부된 도면을 참고하여 본 발명의 실시예를 상세히 설명하기로 한다.
본 발명의 일측면에 따른 옥시토모듈린 생산용 융합태그는 서열번호1, 서열번호2, 서열번호3, 서열번호4, 서열번호 5 및 서열번호6으로 이루어진 군으로부터 선택되는 하나의 아미노산 서열을 포함할 수 있다.
재조합 미생물 시스템을 이용하여 여러가지 목적 펩타이드 또는 목적 단백질을 생산하는 경우, 그 목적 물질의 물성에 따라 숙주 세포 내 효소에 의한 분해, 낮은 발현 수준, 부적절한 단백질 폴딩 및/또는 낮은 mRNA 안정성 등으로 인해 수율이 저하될 우려가 있다. 예를 들어, 종래 융합 태그로 활용되는 MBP (maltose binding protein), 글투타티온 S-전달효소 (glutathione-S-transferase), 티오레독신(thioredoxin), SUMO, 유비퀴틴 등은 각각 397, 216, 106, 101, 76 개의 아미노 산을 갖는데, 비교적 저분자량의 목적 펩타이드 등을 생산 시 수율이 좋지 못한 문제가 있었다.
이에 반해, 본 발명에 따른 상기 융합태그는 상대적으로 분자량이 작은 펩타이드로서, 이를 적용하여 GLP-1, GLP-2 또는 옥시토모듈린 등의 목적 펩타이드 또는 목적 단백질을 생산하는 경우 기존의 융합태그들을 활용한 경우보다 향상된 수율로 목적 펩타이드 또는 목적 단백질을 수득할 수 있다. 예를 들어, 융합태그 및 목적 펩타이드 또는 목적 단백질로 이루어진 재조합 융합 폴리펩타이드에서 옥시토모듈린의 비율을 아래 표 1에 개략적으로 나타내었다.
융합 태그 아미노산 개수 분자량 (kDa) 목적 펩타이드 비율 (%)*
MBP(Maltose binding protein) 397 44.2 8
글루타티온 S-전달효소 216 23.8 12
서열번호 1내지 6의 아미노산 서열을 포함하는 융합태그 78 ~ 281 8.6 ~ 28.8 9 ~ 28
*옥시토모듈린을 기준으로 링커를 포함하여 계산
표 1에 나타난 바와 같이, 종래의 융합태그들은 재조합 융합 폴리펩타이드에서 옥시토모듈린의 비율이 8% 내지 12%에 그치는 반면, 본 발명에 따른 융합태그가 융합된 융합 폴리펩타이드에서의 옥시토모듈린의 비율은 9% 내지 28%로 확인된다. 따라서, 동일한 농도의 융합 폴리펩타이드로부터 얻을 수 있는 옥시토모듈린의 양이 종래 융합태그 대비 더 높으므로 최종 생산 수율이 향상될 수 있다.
또한, 본 발명에 따른 융합태그는 목적 펩타이드 또는 목적 단백질을 포함하는 융합 폴리펩타이드의 불용성 발현을 유도하여 융합 폴리펩타이드가 불용성 내포체로서 숙주세포 내로 고농도로 축적되게 할 수 있다. 따라서, 대장균(Escherichia coli) 등의 숙주세포 내에서 프로테아제 및 펩티다제에 의해 전부 혹은 일부가 분해 혹은 절단될 우려가 있는 목적 펩타이드 또는 목적 단백질을 고수율로 수득하기 용이한 장점이 있다.
본 발명에서 사용된 용어 "목적 펩타이드 또는 목적 단백질"은 재조합 생산 시스템을 통해 수득하고자 하는 펩타이드 또는 단백질을 의미하며, 세포내 효소에 의한 분해, 낮은 발현 수준, 부적절한 단백질 폴딩 및/또는 낮은 mRNA 안정성 등의 다양한 이유로 인해 고수율로 발현되기 어려운 것으로 확인된 펩타이드 또는 단백질이다.
상기 목적 펩타이드 또는 목적 단백질은 본 발명에 따른 융합태그와 융합을 통해, 발현수준이 향상될 뿐만 아니라, 불용성 내포체 형태로 세포 내에 축적됨으로 써 숙주 세포 내 효소에 의한 분해로부터 보호될 수 있어, 결과적으로 더 높은 수율로 수득될 수 있다. 구체적으로, 상기 목적 펩타이드 또는 목적 단백질은 옥신토모듈린(oxyntomodulin, OXM)일 수 있다.
상기 옥신토모듈린(OXM)은 글루카곤의 전구체인 프리-글루카곤(pre-glucagon)으로부터 만들어지는 펩타이드로서, GLP-1의 음식물 섭취 저해, 포만감 증진 효력과 글루카곤의 지방분해 기능을 가질 수 있다. 또한, GLP-1과 글루카곤의 두 펩타이드의 수용체에 모두 결합할 수 있는 이중 기능성을 가질 수 있다.
본 발명의 일실시예에 따르면, 상기 융합태그는 서열번호 1을 포함하는 아미노산을 갖는 펩타이드일 수 있다.
구체적으로, 상기 서열번호 1의 아미노산 서열을 포함하는 융합태그는 Intramolecular chaperone(IMC; 분자내 샤페론)으로, 상기 IMC는 성숙단백질의 안정성이나 활성에는 필요하지 않지만 성숙단백질의 폴딩에는 필수적이며, 성숙단백질의 N-말단에서 생합성되는 수십 내지 수백 개의 결합형 아미노산 프로펩타이드일수 있다. 또한, 상기 융합태그는 다양하게 변형된 펩타이드, 즉 변이체를 포함할 수 있다. 상기 변형은 융합태그의 기능을 변형시키지 않는 내에서 하나 이상의 단백질을 치환, 결실 또는 추가하는 방법을 통하여 수행될 수 있다. 이러한 다양한 펩타이드는 서열번호 1의 아미노산 서열 중 어느 하나와 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 또는 99% 이상 동일한 것일 수 있다.
본 발명의 일실시예에 따르면, 상기 융합태그는 서열번호 2, 서열번호3, 서열번호 4, 서열번호 5 또는 서열번호 6을 포함하는 아미노산을 갖는 펩타이드일 수 있다.
상기 서열번호 2의 아미노산 서열을 포함하는 융합태그는 IMC-Fragment B1으로, 상기 IMC의 변이체일 수 있으며, 상기 서열번호 3의 아미노산 서열을 포함하는 융합태그는 IMC-Fragment B2으로, 상기 IMC의 변이체일 수 있다. 상기 IMC-Fragment B1, B2 (IMCB1, IMCB2)는 각각 서브틸리신(subtilisin) 폴리펩타이드의 51 및 96개의 잔기를 포함할 수 있다. 또한, 서열번호 4의 아미노산 서열을 포함하는 융합태그는 IMC-Fragment C1으로, 상기 IMC의 변이체일 수 있으며, 상기 서열번호 5의 아미노산 서열을 포함하는 융합태그는 IMC-Fragment C2으로, 상기 IMC의 변이체일 수 있다.
상기 IMC-Fragment C1, C2 (IMCC1, IMCC2)는 각각 서브틸리신(subtilisin) 폴리펩타이드의 118 및 146 개의 잔기를 포함할 수 있다.
또한, 서열번호 6의 아미노산 서열을 포함하는 융합태그는 IMC-Fragment D1 (IMCD1)으로, 상기 IMC의변이체일 수 있다. 상기 IMC-Fragment D1은 서브틸리신(subtilisin) 폴리펩타이드의 203개의 잔기를 포함할 수 있다.
본 발명의 일실시예에 따르면, 상기 IMC변이체들은 상기 IMC를 기반으로 서브틸리신 폴리펩타이드가 추가된 것으로서 상기 IMC 및 그 변이체들의 융합태그로서의 효능은 본 발명에서의 실시예들을 살펴볼 때 입증이 되었고, 또한 상기 IMC및 그 변이체들은 그 구조적 연관성으로 인해 어느 하나의 융합태그의 효능이 다른 융합태그에서도 충분히 기대할 수 있다는 점은 당업자에게 자명하다고 할 수 있다.
본 발명의 또 다른 측면은, 전술한 바와 같은 융합태그, 옥시토모듈린 및 상기 융합태그와 상기 옥시토모듈린 사이에 링커를 포함하는 융합 폴리펩타이드를 제공할 수 있다.
상기 링커는 친화성 태그를 포함할 수 있다. 본 발명에서 사용된 용어 "친화성 태그"는 재조합 융합 폴리펩타타이드 또는 이를 코딩하는 핵산에 도입될 수 있는 펩타이드 또는 핵산 서열로서 다양한 목적으로 사용될 수 있으며, 예를 들어 목적 펩타이드 또는 목적 단백질의 정제 효율을 높이기 위한 것일 수 있다. 본 발명에서 사용될 수 있는 친화성 태그는 당 업계에 공지된 임의의 적합한 물질을 원하는 바에 따라 사용할 수 있다. 예를 들어, 본 발명에 사용되는 친화성 태그는 폴리히스티딘 태그(서열번호8)일 수 있다.
또한, 상기 링커는 프로테아제 인식 서열을 포함할 수 있다. 프로테아제란, 특정 아미노산 서열을 인식하여, 인식한 서열 내의 펩타이드 결합이나 그 서열의 마지막 아미노산과 융합된 폴리펩타이드의 첫번째 아미노산과의 펩타이드 결합을 절단하는 단백질 분해효소이다. 본 발명에 따른 융합 폴리펩타이드는 프로테아제 인식 서열을 갖는 링커를 포함함으로써, 최종 단계에서 폴리 펩타이드 정제 시 절단 효소 인식 서열이 포함된 아미노 말단(친화성 태그를 사용한 경우에는 친화성 태그도 포함)과 옥시토모듈린의 N-말단을 분리시켜 옥시토모듈린을 회수할 수 있다.
구체적으로, 상기 프로테아제 인식 서열은 담배 식각 바이러스(TEV) 프로테아제 인식 서열, 엔테로키나아제 인식 서열, 유비퀴틴 가수분해효소 인식 서열, 인자Xa, 퓨린 및 이들의 조합으로 이루어진 군으로부터 선택되는 인식 서열일 수 있다. 바람직하게, 상기 프로테아제 인식 서열은 서열번호9의 아미노산을 포함할 수 있다.
본 발명의 다른 측면은, 전술한 융합태그; 및 옥시토모듈린을 코딩하는 뉴클레오타이드를 제공할 수 있다.
본 발명의 또 다른 측면은, 전술한 전술한 융합태그; 및 옥시토모듈린을 코딩하는 뉴클레오타이드를 포함하는 발현 벡터를 제공할 수 있다. 본 발명의 일실시예에 따르면, 상기 발현벡터는 Trc 프로모터, lac 오퍼레이터, 히스티딘태그 코딩 서열, T7 터미네이터, rrnB 터미네이터, 카나마이신 저항 서열, pBR 오리진, lacl q서열을 포함할 수 있다(도 1). 본 발명의 다른 일실시예에 따르면, 상기 발현벡터는 서열번호 171의 염기서열을 포함할 수 있다.
본 발명에서 사용된 용어 "벡터"는 숙주 세포에 도입되어 숙주 세포 유전체 내로 재조합 및 삽입될 수 있다. 상기 벡터는 선형 핵산, 플라스미드, 파지미드, 코스미드, RNA 벡터, 바이러스 벡터 및 이의 유사체들을 포함한다. 바이러스 벡터의 예로는 레트로바이러스, 아데노바이러스, 및 아데노-관련 바이러스를 포함할 수 있다. 또한, 상기 플라스미드는 항생제 내성 유전자와 같은 선별마커를 포함할 수 있고, 플라스미드를 유지하는 숙주 세포는 선택적인 조건하에서 배양될 수 있다.
또한, 본 발명의 융합태그 및 옥시토모듈린을 코딩하는 뉴클레오타이드 서열을 포함하는 발현 벡터는 유도성 발현 벡터의 일부로서 제공될 수 있다. 예를 들어, 발현벡터로 형질전환된 숙주 세포를 배양하고, 발현 벡터로부터 융합 단백질의 발현을 유도한다. 발현 벡터는 플라스미드일 수 있다. 예를 들어, 발현 벡터는 재조합 융합 단백질을 암호화하고 있는 유전자와 선별 마커에 대한 유전자를 추가로 포함하는 플라스미드이고, 숙주 세포는 플라스미드를 유지하도록 하는 선별 조건하에서 배양된다.
상기 발현 벡터는 단백질 코딩 서열 이외에 프로모터, 리보솜 결합 부위(RBS), 전사 종결 인자, 및 번역 개시 및 정지 신호 등의 단백질 발현과 관련된 조절 요소를 포함할 수 있다. 예를 들어, 유용한 목적 펩타이드의 발현 수준에 영향을 주는 RBS는 다양한 생물의 유전자나 합성을 통해 얻을 수 있다.
또한, 본 발명에서 사용되는 프로모터는 조절되는 유도성 프로모터로서, 예를 들어, trc 프로모터, lac 프로모터, tac 프로모터 및 trc 프로모터로부터 유래한 프로모터일 수 있다. 구체적으로, trc 프로모터는 일반적으로 lac 계열 프로모터를 사용하는 발현 시스템에는 발현 조절자로서 lacI 유전자가 발현 시스템에 존재할 수 있다. 보통 항시적으로 발현되는 유전자인 lacI 유전자는 lac 패밀리 프로모터의 lac 작동 인자(operator)에 결합하는 Lac 억제인자 단백질 LacI 단백질을 코딩한다. 따라서, lac 계열 프로모터가 이용되는 경우, lac 유전자는 또한 발현시스템에 포함되고 발현될 수 있다.
본 발명에서 사용된 용어, 목적 단백질의 "유전자 발현" 또는 "발현"은, DNA서열의 전사, mRNA 전사체의 번역 및 융합 단백질 생산물 또는 이의 단편의 분비를 의미하는 것으로 이해된다.
본 발명에서 사용된 용어 "숙주 세포"는 재조합 발현 벡터가 도입될 수 있는 원핵 및 진핵 세포를 나타낸다. 본 발명에서 사용된 용어, "형질주입된", "형질전환된" 및 "형질감염된"은 당업계에 공지된 기술을 사용하여 세포 내로 핵산(예를 들어, 벡터)을 도입하는 것을 의미한다.
본 발명에서 사용된 용어, 목적 단백질의 "유전자 발현" 또는 "발현"은, DNA서열의 전사, mRNA 전사체의 번역, 및 융합 단백질 생산물을 의미한다.
본 발명의 다른 측면은, 상기 발현 벡터를 포함하는 숙주 세포를 제공할 수 있다. 적절한 숙주 세포는, 본 발명의 DNA 서열이 형질주입 또는 형질감염되어, 목적 펩타이드 또는 목적 단백질의 발현 및/또는 분비에 이용될 수 있으며, 본 발명에 사용될 수 있는 바람직한 숙주 세포는 대장균 세포, 불사의 하이브리도마 세포, NS/0 골수종 세포, 293 세포, 중국 햄스터 난소 세포(CHO cell), HeLa 세포, 인간 양수 유래 세포(CapT cell) 또는 COS 세포를 포함한다. 예를 들어, 본 발명에서 융합 폴리펩타이드를 발현시키는데 사용된 숙주 균주는 대장균 BL21(DE3)이며, 상기 유전자 및 이들의 사용 방법은 당 업계에 공지되어 있다.
또한, 재조합 융합 펩타이드 또는 단백질을 발현하기 위한 최적의 조건을 결정하기 위해 고처리량 스크리닝(high throughput screening)을 수행할 수 있다. 스크리닝에서 변화될 수 있는 조건은, 숙주 세포, 숙주 세포의 유전적 배경 (예를 들어, 다양한 프로테아제와 같은 특정 유전자의 결실), 발현 구조물 내 프로모터의 유형, 발현 구조물 내 RBS의 유형, 배양 온도, 유도성 프로모터가 사용될 때 유도 시 세포 탁도, lac 계열 프로모터가 사용될 때 유도에 사용되는 IPTG의 농도, 단백질 유도기간, 배양물에 유도제 첨가 후 배양 온도, 배양물의 교반 속도, 플라스미드 유지에 대한 선별 방법, 용기 내 배양물의 부피, 및 세포 용해 방법을 포함한다.
또한, "최적의 숙주 균주" 또는 "최적의 발현 시스템"은 유전형(genotype)이 상이한 다양한 숙주 세포와 비교하여 발현된 재조합 융합 단백질의 품질, 발현 양, 발현 수준 및/또는 위치를 기준으로 하여 확인하거나 선택할 수 있다. 따라서, 최적의 숙주 균주는 목적 펩타이드 또는 목적 단백질의 생산에 적합하도록 재조합 융합 단백질을 생산하는 균주이다. 목적 펩타이드 또는 목적 단백질의 생산에 대한 높은 적합성은 생산되는 목적 펩타이드 또는 목적 단백질에 따라 달라질 수 있지만 재조합 융합 단백질의 품질, 발현 양, 발현 수준, 예를 들어 목적 펩타이드 또는 목적 단백질의 세포 내 분해 혹은 부분적인 절단 유무가 발생되었는지, 어느 정도의 발현양 혹은 발현 수준인지, 단백질이 적합하게 또는 바람직하게 폴딩되거나 불용성 내포체로 축적되었는지 등을 포함한다. 즉, 최적의 숙주 균주 또는 최적의 발현 시스템은 대조군 숙주 균주나 발현 시스템과 비교하여 특정 수준 이상의 생산 수율, 발현 양, 발현 수준, 회수 가능한 재조합 융합 단백질의 양, 적합하게 불용성 내포체 형태로 축적된 재조합 융합 단백질의 양을 생산할 수 있다.
이하, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 본 발명의 실시예에 대하여 상세히 설명한다. 그러나 본 발명은 여러 가지 상이한 형태로 구현될 수 있으며 여기에서 설명하는 실시예에 한정되지 않는다.
실시예 1. 융합태그 및 옥시토모듈린 발현을 포함하는 융합 폴리펩타이드 생산을 위한 pSK03플라스미드 제작
본 발명에서 융합태그 및 옥시토모듈린을 포함하는 융합 폴리펩타이드의 재조합 생산을 위한 발현 플라스미드인 pSK03은 pTrcHis2A (Invitrogen), pET26b (Novagen) 및 pGNX4 (삼양제넥스)의 유전자들을 조합하여 제작되었다. 아래 표 2의 프라이머를 이용하여 각 유적자 부위를 PCR로 증폭 후 In-fusion 클로닝 방법을 이용하여 pSK01과 pSK02를 제작한 후 pSK03을 최종 제작하였다. PCR 증폭은 Pfu DNA 중합효소를 사용하여 94℃ 5분 이후 94℃ 1분, 55℃ 1분 및 72℃ 5분에서 30 사이클을 반복하고 5분 72℃ 10분 조건에서 마무리하여 실시하였다. 증폭된 유전자는 Dpn I 제한효소를 이용하여 메틸기를 제거하고, T4 DNA 중합효소를 이용하여 유전자 절편을 연결하였다. 또한 Pfu DNA 중합효소는 바이오니아, T4 DNA 중합효소는 NEB에서 구입하였다.
프라이머 번호 서열번호 프라이머 명칭 DNA 서열
1 서열번호 80 lacI-5UTR-F cgtgactgggtcatggctgcg
2 서열번호 81 rrnB-term-IF-R aaggcccagtctttcgactga
3 서열번호 82 rrnB-term-IF-F tcagtcgaaagactgggcctt
4 서열번호 83 lacI-5UTR-R cgcagccatgacccagtcacg
5 서열번호 84 pSG01-pelB-IF-F aataaggaggaataacatatgaaatacctgctgccg
6 서열번호 85 6his-pSG01-IF-R gaccgtttaaactcagtggtggtggtggtggtgctc
7 서열번호 86 pSK01-IF-F tgagtttaaacggtctccagc
8 서열번호 87 pSK01-IF-R cgcagccatgacccagtcacg
9 서열번호 88 T7ter-rrnBter-F gtcttgaggggttttttgagcggtctgataaaacag
10 서열번호 89 lacO-R gggaattgttatccgctcacaattcc
11 서열번호 90 lacO-F ggaattgtgagcggataacaattccc
12 서열번호 91 T7ter-rrnBter-R ctgttttatcagaccgctcaaaaaacccctcaagac
실시예 1.1. pSK01 플라스미드의 제작
pTrcHis2A의 lac 억제자, trc 프로모터, lac 작동자(operator), 리보솜 결합부위 (RBS) 및 rrn 터미네이터 유전자를 포함하는 절편은 프라이머 1(서열번호 80)과 프라이머 2(서열번호 81)를 사용하고 pGNX4의 rrnB 터미네이터, 카나마이신 내성유전자, pBR322 복제원점의 자리가 포함된 절편은 프라이머 3(서열번호 82)과 프라이머 4(서열번호 83)을 사용하여 PCR 증폭을 통해 획득하였다. 상기 2개의 DNA는 PCR purification kit를 이용하여 해당 DNA만 정제하였다. 그 결과물인 두개의 DNA 절편을 T4 DNA 중합효소를 사용하여 연결시켰고 대장균 MC1061세포에 형질전환 시켰다. 형질전환체 내의 pSK01 플라스미드를 추출 후 DNA 염기서열 분석을 통해 상기 2 두개의 DNA가 정확하게 연결된 것을 확인하였다.
실시예 1.2. pSK02 플라스미드 제작
상기 과정을 통해 제작된 pSK01의 다중 클로닝 부위 (Multiple Cloning Site)가 pET26b의 PelB 신호서열 (signal peptide)과 다중 클로닝 부위로 대체된 pSK02 플라스미드를 다음 방법으로 제작하였다. pSK01의 다중 클로닝 부위를 제외한 나머지 유전자 부분를 증폭하기 위해 프라이머 5(서열번호 84)와 프라이머 6(서열번호 85)을 사용하였고 pET26b의 PelB 신호서열과 다중 클로닝 부위를 증폭하기 위해 프라이머 7(서열번호 86)과 8(서열번호 87)을 사용하였다. PCR을 통해 증폭된 상기 두 DNA는PCR purification kit를 이용하여 해당 DNA만 정제하였다. 그 결과물인 두개의 DNA 절편을 T4 DNA 중합효소를 사용하여 연결시켰고 대장균 MC1061세포에 형질전환 시켰다. 형질전환체 내의 pSK02 플라스미드를 추출 후 DNA 염기서열 분석을 통해 pSK01의 다중 클로닝 부위가 제거되고 pET26b의 PelB 신호서열과 다중 클로닝 부위가 정확히 삽입된 것을 확인하였다.
실시예 1.3. pSK03 플라스미드 제작
상기 과정을 통해 제작된 pSK02의 다중 클로닝 부위와 rrn 터미네이터 유전자 사이에 T7 터미네이터 유전자가 삽입된 pSK03플라스미드를 다음 방법으로 제작하였다. 프라이머 9(서열번호 88)와 프라이머 10(서열번호 89)을 사용하여 pSK02의 Iac 작동자(operator) 부터 다중 클로닝 부위를 제외한 나머지 유전자 부분를 증폭하였다. pET26 내 Iac 작동 인자(operator), 리보솜 결합부위(RBS), pelB signal peptide, T7 터미네이터를 포함하는 유전자를 프라이머 11(서열번호 90)과 프라이머 12(서열번호 91)를 사용하여 증폭하였다. PCR을 통해 증폭된 상기 두 DNA는PCR purification kit를 이용하여 DNA만 정제하였다. 그 결과물인 두개의 DNA 절편을 T4 DNA 중합효소를 사용하여 연결시켰고 대장균 MC1061세포에 형질전환 시켰다. 형질전환체 내의 pSK03 플라스미드를 추출 후 DNA 염기서열 분석을 통해 pSK03 플라스미드는 다중 클로닝 부위와 rrn 터미네이터 유전자 사이에 T7 터미네이터 유전자가 정확히 삽입된 것을 확인하였다. 도 1은 pSK03 플라스미드의 모식도를 나타낸다.
실시예 2. OXM 융합 폴리펩타이드의 제조 및 생산
실시예 2.1. OXM 융합 폴리펩타이드 발현 플라스미드의 제작
OXM융합 폴리펩타이드에 대한 유전자는 overlap extension polymerase chain reaction (OE-PCR) 방법을 이용하여 합성하였다. 이때, 상기hGH융합 태그는 아미노 말단 융합태그로서 IMC(서열번호 1), IMC-Fragment B1 (IMCB1, 서열번호 2), IMC-Fragment B2 (IMCB2, 서열번호 3), IMC-Fragment C1 (IMCC1, 서열번호 4), IMC-Fragment C2 (IMCC2, 서열번호 5), IMC-Fragment D1 (IMCD1, 서열번호 6) 중 하나와6 히스티딘 태그(서열번호 8) 및 TEV 프로테아제 인식 서열(서열번호 9) 및 OXM의 아미노산 서열(서열번호 71)이 포함되어 있다.
음성 대조군으로서 사용된 OXM 융합 폴리펩타이드(H6TEV-OXM, 서열번호 72)는 아미노 말단 융합태그가 포함되어 있지 않고, 6 히스티딘 태그(서열번호 8), TEV 프로테아제 인식 서열(서열번호 9) 및 OXM의 아미노산 서열(서열번호71)이 포함되어 있다.
양성 대조군으로 사용된 OXM 융합 폴리펩타이드(SUMO-H6TEV-OXM, 서열번호 79)는 아미노 말단 융합태그 SUMO(서열번호 7) 와 6 히스티딘 태그(서열번호 8) 및 TEV 프로테아제 인식 서열(서열번호 9) 및 OXM의 아미노산 서열(서열번호 71)이 포함되어 있다.
각 융합 폴리펩타이드의 유전자는 제한 효소 NdeI 및 XhoI의 인식 서열과 1개의 종결 코돈을 포함한다. 상기 OXM융합 폴리펩타이드를 코딩하는 뉴클레오타이드 서열은 서열번호 164 내지 169에 해당하고 아미노산 서열은 서열번호 73 내지 78에 해당한다. 또한, 음성 및 양성 대조군의 뉴클레오타이드 서열은 서열번호 163및 서열번호 170에 해당하고 아미노산 서열은 서열번호 72 및 서열번호 79에 해당한다.
아래 표3에 기재한 OXM 융합 폴리펩타이드 발현 플라스미드 pSGK850, pSGK763, pSGK797, pSGK798, pSGK799, pSGK800, pSGK801를 제작하기 위해서, OE-PCR로 합성된 OXM 융합단백질 유전자 단편을 NdeI 및 XhoI 제한 효소를 이용하여 절단하고, trc 프로모터, lac 작동자 및 lacI 유전자를 포함하여 IPTG에 의한 발현 조절이 가능한 발현 벡터pSK03에 클로닝하였다.
균주 숙주세포 플라스미드 재조합 융합 폴리펩타이드
PG095 E. coli BL21(DE3) pSGK797 H6TEV-OXM
PG096 E. coli BL21(DE3) pSGK798 SUMO-H6TEV-OXM
PG097 E. coli BL21(DE3) pSGK799 IMCB1-H6TEV-OXM
PG098 E. coli BL21(DE3) pSGK800 IMCB2-H6TEV-OXM
PG099 E. coli BL21(DE3) pSGK801 IMCC1-H6TEV-OXM
PG100 E. coli BL21(DE3) pSGK802 IMCC2-H6TEV-OXM
PG101 E. coli BL21(DE3) pSGK803 IMCD1-H6TEV-OXM
제작된 OXM 융합 폴리펩타이드 발현 플라스미드들은 DNA 염기서열 확인을 통해 정확한 클로닝 여부를 확인하였다. 제작된 OXM 융합 폴리펩타이드 발현 플라스미드들을 이용하여 염화칼슘을 이용한 화학적인 방법으로 대장균 BL21(DE3) 세포를 형질전환하였다. OXM 융합 폴리펩타이드의 발현 플라스미드들이 형질도입된 대장균들은 카나마이신(kanamycin)이 50 ㎍/㎖의 농도로 포함된 LB 고체배지에서 콜로니를 형성하였다. 형질전환된 대장균들은 각각 카나마이신이 50 ㎍/㎖의 농도로 존재하는 LB 액체 배지에 배양한 후, 50% 글리세롤을 배양액과 동일한 부피로 첨가하여 세포 농축액(cell stock)을 만들고, 이를 -80℃ 온도의 냉동고에 보관하였다.
실시예 2.2. 형질전환된 세포의 배양 및 OXM의 발현
-80℃ 온도에서 보관된 OXM 융합 폴리펩타이드의 발현 플라스미드들이 형질전환된 대장균의 세포 농축액을 실온에서 녹인 후 50 ㎕를 카나마이신이 50 ㎍/㎖의 농도로 존재하는 LB 액체 배지 5 ㎖이 포함된 시험관에 첨가하여 37℃ 온도의 진탕배양기에서 12시간 동안 종균 배양하였다. 종균 배양된 대장균 2 ㎖는 카나마이신이 50 ㎍/㎖의 농도로 존재하는 LB 액체 배지 200 ㎖이 포함된 플라스크에 첨가하여 37℃ 온도의 진탕배양기에서 배양하였다. 배양 약 3시간 후 세포의 광학밀도(OD600)가 약 1.0이 되었을 때, IPTG의 최종 농도가 0.1 mM이 되도록 첨가하여 OXM 융합 폴리펩타이드의 발현을 유도하였다. 발현 유도 4시간 후에 광학 밀도를 측정하여 세포의 광학 밀도를 측정하였다.
실시예 2.3. 발현 수준 비교 분석을 위한 시료의 제조
발현 유도가 된 세포의 광학밀도가 10.0이 되도록 세포를 농축하고 50 mM 인산 나트륨, pH 7.2 완충액으로 재현탁시킨 후 초음파 세포 파쇄기(ultrasonic processor, Cole-Parmer)를 이용하여 세포를 파쇄하였다. 파쇄된 세포는 총 세포 분획으로 표지하였다. 세포 파쇄액은 12,000×g, 4℃ 온도에서 15분간 원심분리하였다. 상등액을 회수하고 가용성 분획으로 표지하였다. 불용성 분획은 500 ㎕의 50 mM 인산 나트륨, pH 7.2 완충액으로 초음파 세포 파쇄기를 이용하여 재현탁하고 불용성 분획으로 표지하였다.
실시예 2.4. SDS-PAGE 분석을 통한 생산된 OXM 융합 폴리펩타이드의 확인
50 ㎕의 총 세포, 가용성 및 불용성 분획을 각각 50 ㎕의 2배 농축된 SDS 시료 완충액(SDS sample buffer 2× concentrate, Sigma)에 혼합한 후, 95℃ 온도에서 5분간 가열하여 각 시료의 단백질이 변성될 수 있도록 하였다. 시료 내 변성된 단백질들은 16% SDS-PAGE 겔 및 TANK 완충액을 이용하여 분자량 크기에 따라 겔 내에서 분리될 수 있도록 하였다. SDS-PAGE 후 겔을 쿠마시 블루(Coomassie blue) R-250이 포함된 염색 완충액으로 염색한 후, 탈색 완충액으로 탈색하여 염색된 단백질만 보일 수 있도록 하였다.
도2을 참조하면, 음성 대조군인 융합태그가 포함되지 않은 H6TEV-OXM(6.2 kDa의 분자량)는 타 OXM융합 폴리펩타이드들과 비교하여 가장 낮은 발현 수준을 보였다. H6TEV-OXM의 아미노 말단에 IMCD1이 융합된 IMCD1-H6TEV-OXM(34.9 kDa의 분자량)의 발현 수준은 H6TEV-OXM대비 매우 향상된 것이 확인되었다. 또한 양성 대조군인 SUMO-H6TEV-OXM(17.3 kDa의 분자량)와 비교해도 IMCD1-H6TEV-OXM의 발현수준이 월등히 높은 것으로 확인되었다. IMCD1-H6TEV-OXM은 재조합 대장균에서 불용성 내포제 형태로 발현되는 것을 확인되었다(도3).
서열번호 2의 아미노 말단 융합태그인 IMCB1이 융합된 OXM 융합 폴리펩타이드인 IMCB1-H6TEV-OXM(19.9 kDa의 분자량)은 IMCD1-H6TEV-OXM과 유사한 발현 수준을 보였다(도4).
실시예 3. IMCB1-H6TEV-OXM의 회수 및 정제
실시예 3.1. 세포 파쇄 및 불용성 내포체 회수
플라스크 규모에서 발현된 세포의 냉동된 세포 펠렛을 50 ㎖의 50 mM 인산 나트륨, pH 7.0 완충액을 첨가하여 해동하였다. 재현탁된 세포는 초음파 세포 파쇄기(ultrasonic processor, Cole-Parmer)를 이용하여 파쇄하였다. 용해된 세포는 12,000 rpm(12,000×g) 조건에서 30분간 원심분리하고 상등액을 제거하여 재조합 융합 폴리펩타이드가 포함된 불용성 내포체 분획을 회수하였다.
실시예 3.2. 불용성 내포체 가용화
회수된 불용성 내포체 분획에 20 ㎖의 내포체 가용화 완충액(6 M 구아니딘, 20 mM 인산나트륨, 염화나트륨, 20 mM 이미다졸, pH 7.4)을 첨가하고 4℃ 온도에서 1시간 동안 진탕배양하여 불용성 분획 내 내포체 형태의 재조합 융합 폴리펩타이드가 가용화될 수 있도록 하였다. 가용화된 불용성 분획 시료는 12,000Х에서 30분간 원심분리하고 상등액을 막필터(0.45/0.2 ㎛)를 통해 여과하였다.
실시예 3.3. IMCB1-H6TEV-OXM의 정제
가용화된 불용성 분획 내의 IMCB1-H6TEV-OXM의 위하여 S9 시료 펌프(Sample pump S9) 및 F9-C 분획 수집기(Fraction collector F9-C)이 장착된 AKTA pure 25 크로마토그라피 시스템(GE Healthcare)을 이용하였다. 가용화된 불용성 분획 시료는 내포체 가용화 완충액(8 M 요소, 20 mM 트리스, 500 mM 염화나트륨, 50 mM 이미다졸, pH 7.4)으로 미리 평형화된 HisTrap FF 1 ㎖ 컬럼(GE Healthcare)에 주입하였다. 주입이 완료된 후 컬럼은 5배 컬럼 부피의 평형 완충액으로 세척하고, 5배 컬럼 부피의 용출 완충액(8 M 요소, 20 mM 트리스, 500 mM 염화나트륨, 500 mM 이미다졸, pH 7.4)을 단계적으로 100%가 되도록 증가시켜 컬럼의 수지에 결합되어 있는 IMCB1-H6TEV-OXM을 용출하였다. 용출 후 얻어진 분획의 결과를 각각 도5에 나타내었다.
실시예 4. 프로테아제 처리를 통한 링커 서열 절단
약 5 ㎖의 정제된 IMCB1-H6TEV-OXM 융합 폴리펩타이드의 분획을 합친 후 희석 완충액(50 mM 트리스, pH 8.0)을 첨가하여 구아니딘의 농도가 1 M이 되도록 희석하였다. 희석된 재조합 융합 폴리펩타이드에 TEV 프로테아제의 최종 농도가 500 nM이 되도록 첨가하고 실온에서 12시간 동안 절단 반응이 진행될 수 있도록 하였다.
TEV 프로테아제에 의한 절단 여부를 확인하기 위해, 절단 후 SDS-PAGE 분석을 수행하여 그 결과를 도6에 나타내었다. 한편, 도6을 참조하면, 19.9 kDa의 IMCB1-H6TEV-OXM는 거의 100%의 수율로 N-말단 융합태그, 6 히스티딘 태그 및 TEV 프로테아제 인식서열 융합체인 IMCB1-H6TEV 단편(15.5 kDa)과 목적 펩타이드인 OXM(4.4 kDa)단편으로 분리되는 것을 확인하였다.
실시예 5. OXM의 정제
IMCB1-H6TEV-OXM 융합 폴리펩타이드에서, TEV 프로테아제의 절단에 의해 유리된 GLP-1K28R의 정제를 위하여 S9 시료 펌프(Sample pump S9) 및 F9-C 분획 수집기(Fraction collector F9-C)이 장착된 AKTA pure 25 크로마토그라피 시스템(GE Healthcare)을 이용하였다. TEV 프로테아제로 절단된 각각의 시료를 결합 완충액(10 mM 인산 칼륨, 2% 아세톤니트릴 pH 7.0)으로 투석하여 완충액 교환 후 동일 완충액으로 미리 평형화된 Resource RPC ㎖ 컬럼(GE Healthcare)에 주입하였다. GLP-1K28R은 컬럼에 결합되지 않으므로 통과액(Flow-through) 분획을 수집하였고 용출 완충액(10 mM 인산 칼륨, 70% 아세톤니트릴 pH 7.0)을100%가 되도록 컬럼에 통과시켜 컬럼의 수지에 결합되어 있는IMCB1-H6TEV및 타 불순물들을 제거하였다. 분획 수집기의 통과액 분획 및 OXM을 포함하는 용출 분획들은 상기에 기재된 SDS-PAGE 방법으로 분석하였다.
Resource RPC수지에 결합된IMCB1-H6TEV및 타 불순물들은 낮은 농도의 아세톤니트릴에서 용출되고 OXM은 높은 농도의 아세톤니트릴에 의해 용출되는 것을 확인하였다. 아세톤니트릴의 농도를 조절함으로써 OXM을 용이하게 분리 가능하므로 역상 크로마토그래피를 이용하여OXM을 고순도로 정제할 수 있다. 분획 수집기의 통과액 및 용출 분획은 상기에 기재된 SDS-PAGE 방법으로 분석하였다.
도7를 참조하면, OXM은 Resource RPC수지에 결합된 후 고농도의 아세톤니트릴에 의해 용출되는 것을 확인하였다. 따라서 IMCB1-H6TEV와 OXM은 아세톤니트릴의 농도를 조절함으로써 용이하게 분리 가능하므로 역상 크로마토그래피를 이용하여 OXM의 분리 정제를 용이하게 할 수 있다.
실시예 6. 절단 후 정제된 OXM의 분자량 분석
완전한 형태의 IMCB1-H6TEV-GLP-OXM의 발현 유무, TEV 프로테아제에 의한 정확한 절단 및 절단 후 얻어진 OXM의 변형(modification) 유무를 확인하기 위해 MALTI-TOF MS를 이용한 분자량 분석을 실시하였다. 본 발명에 따라 수득된 OXM의 분자량을 측정한 결과를 도8에 나타내었다.
도8을 참고하면, IMCB1-H6TEV-OXM으로부터 수득된 OXM의 분자량은 4450.0 Da으로 측정되어 이론적인 분자량과 동일하였고 대장균 내에서 단백질 분해효소(proteolytic enzyme)에 의한 아미노 혹은 카르복시 말단의 부분적인 절단이나 분해 없이 완전한 형태로 발현된 것을 확인할 수 있었다. 따라서 TEV 프로테아제는 IMCB1-H6TEV-OXM내 인식서열인 ENLYFQ 서열을 인식하고 마지막 아미노산인 Q(글루타민)와 OXM의 첫번째 아미노산인 H(히스티딘) 사이의 펩타이드 결합을 정확하게 절단하는 것으로 판단된다.
실시예 7. 정제된 OXM의 역상 HPLC 분석
본 발명을 통해 생산된 재조합 OXM을 역상 HPLC로 분석한 결과 재조합 OXM의 순도는 95% 이상으로 확인되었다.
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다. 예를 들어, 단일형으로 설명되어 있는 각 구성 요소는 분산되어 실시될 수도 있으며, 마찬가지로 분산된 것으로 설명되어 있는 구성 요소들도 결합된 형태로 실시될 수 있다.
본 발명의 범위는 후술하는 특허청구범위에 의하여 나타내어지며, 특허청구범위의 의미 및 범위 그리고 그 균등 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.
<110> KIM, SUNG GUN <120> FUSION TAG FOR PREPARING OXYNTOMODULIN <130> P20200483KR <150> KR 10-2019-0161763 <151> 2019-12-06 <160> 171 <170> KoPatentIn 3.0 <210> 1 <211> 78 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence for N-terminal fusion partner (IMC) <400> 1 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr 65 70 75 <210> 2 <211> 129 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence for N-terminal fusion partner (IMC-Fragment B1) <400> 2 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val <210> 3 <211> 174 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence for N-terminal fusion partner (IMC-Fragment B2) <400> 3 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu 165 170 <210> 4 <211> 196 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence for N-terminal fusion partner (IMC-Fragment C1) <400> 4 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn 195 <210> 5 <211> 224 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence for N-terminal fusion partner (IMC-Fragment C2) <400> 5 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 <210> 6 <211> 281 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence for N-terminal fusion partner (IMC-Fragment D1) <400> 6 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val 275 280 <210> 7 <211> 98 <212> PRT <213> Artificial Sequence <220> <223> SUMO <400> 7 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly <210> 8 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> Histidine 6 <400> 8 His His His His His His 1 5 <210> 9 <211> 6 <212> PRT <213> Artificial Sequence <220> <223> TEV protease recognition sequence <400> 9 Glu Asn Leu Tyr Phe Gln 1 5 <210> 10 <211> 12 <212> PRT <213> Artificial Sequence <220> <223> H6TEV <400> 10 His His His His His His Glu Asn Leu Tyr Phe Gln 1 5 10 <210> 11 <211> 90 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV <400> 11 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln 85 90 <210> 12 <211> 141 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV <400> 12 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln 130 135 140 <210> 13 <211> 186 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV <400> 13 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln 180 185 <210> 14 <211> 208 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV <400> 14 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 <210> 15 <211> 236 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV <400> 15 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln 225 230 235 <210> 16 <211> 293 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV <400> 16 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln 290 <210> 17 <211> 31 <212> PRT <213> Artificial Sequence <220> <223> Liraglutide precursor peptide (GLP-1K28R) <400> 17 His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly 1 5 10 15 Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Arg Gly Arg Gly 20 25 30 <210> 18 <211> 44 <212> PRT <213> Artificial Sequence <220> <223> H6TEV-GLP-1K28R <400> 18 Met His His His His His His Glu Asn Leu Tyr Phe Gln His Ala Glu 1 5 10 15 Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala 20 25 30 Lys Glu Phe Ile Ala Trp Leu Val Arg Gly Arg Gly 35 40 <210> 19 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV-GLP-1K28R <400> 19 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln His Ala Glu Gly Thr Phe 85 90 95 Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe 100 105 110 Ile Ala Trp Leu Val Arg Gly Arg Gly 115 120 <210> 20 <211> 172 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV-GLP-1K28R <400> 20 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln His Ala Glu 130 135 140 Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala 145 150 155 160 Lys Glu Phe Ile Ala Trp Leu Val Arg Gly Arg Gly 165 170 <210> 21 <211> 217 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV-GLP-1K28R <400> 21 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln His Ala Glu Gly Thr Phe 180 185 190 Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe 195 200 205 Ile Ala Trp Leu Val Arg Gly Arg Gly 210 215 <210> 22 <211> 239 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-GLP-1K28R <400> 22 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly 210 215 220 Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Arg Gly Arg Gly 225 230 235 <210> 23 <211> 267 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV-GLP-1K28R <400> 23 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln His Ala Glu Gly 225 230 235 240 Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys 245 250 255 Glu Phe Ile Ala Trp Leu Val Arg Gly Arg Gly 260 265 <210> 24 <211> 324 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV-GLP-1K28R <400> 24 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser 290 295 300 Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val 305 310 315 320 Arg Gly Arg Gly <210> 25 <211> 141 <212> PRT <213> Artificial Sequence <220> <223> SUMO-H6TEV-GLP-1K28R <400> 25 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly His His His His His His Glu Asn Leu Tyr Phe Gln His Ala 100 105 110 Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly Gln Ala 115 120 125 Ala Lys Glu Phe Ile Ala Trp Leu Val Arg Gly Arg Gly 130 135 140 <210> 26 <211> 34 <212> PRT <213> Artificial Sequence <220> <223> hPTH1-34 (Teriparatide) <400> 26 Ser Val Ser Glu Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn 1 5 10 15 Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His 20 25 30 Asn Phe <210> 27 <211> 47 <212> PRT <213> Artificial Sequence <220> <223> H6TEV-hPTH1-34 <400> 27 Met His His His His His His Glu Asn Leu Tyr Phe Gln Ser Val Ser 1 5 10 15 Glu Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu 20 25 30 Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe 35 40 45 <210> 28 <211> 124 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV-hPTH1-34 <400> 28 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln Ser Val Ser Glu Ile Gln 85 90 95 Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 100 105 110 Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe 115 120 <210> 29 <211> 175 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV-hPTH1-34 <400> 29 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln Ser Val Ser 130 135 140 Glu Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu 145 150 155 160 Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe 165 170 175 <210> 30 <211> 220 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV-hPTH1-34 <400> 30 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln Ser Val Ser Glu Ile Gln 180 185 190 Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg Val Glu 195 200 205 Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe 210 215 220 <210> 31 <211> 242 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-hPTH1-34 <400> 31 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 Ser Val Ser Glu Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn 210 215 220 Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His 225 230 235 240 Asn Phe <210> 32 <211> 270 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV-hPTH1-34 <400> 32 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln Ser Val Ser Glu 225 230 235 240 Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met Glu Arg 245 250 255 Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe 260 265 270 <210> 33 <211> 327 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV-hPTH1-34 <400> 33 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln Ser Val Ser Glu Ile Gln Leu Met His Asn Leu 290 295 300 Gly Lys His Leu Asn Ser Met Glu Arg Val Glu Trp Leu Arg Lys Lys 305 310 315 320 Leu Gln Asp Val His Asn Phe 325 <210> 34 <211> 144 <212> PRT <213> Artificial Sequence <220> <223> SUMO-H6TEV-hPTH1-34 <400> 34 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly His His His His His His Glu Asn Leu Tyr Phe Gln Ser Val 100 105 110 Ser Glu Ile Gln Leu Met His Asn Leu Gly Lys His Leu Asn Ser Met 115 120 125 Glu Arg Val Glu Trp Leu Arg Lys Lys Leu Gln Asp Val His Asn Phe 130 135 140 <210> 35 <211> 33 <212> PRT <213> Artificial Sequence <220> <223> Teduglutide (GLP-2A2G) <400> 35 His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn 1 5 10 15 Leu Ala Ala Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr 20 25 30 Asp <210> 36 <211> 46 <212> PRT <213> Artificial Sequence <220> <223> H6TEV-GLP-2A2G <400> 36 Met His His His His His His Glu Asn Leu Tyr Phe Gln His Gly Asp 1 5 10 15 Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn Leu Ala Ala 20 25 30 Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr Asp 35 40 45 <210> 37 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV-GLP-2A2G <400> 37 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln His Gly Asp Gly Ser Phe 85 90 95 Ser Asp Glu Met Asn Thr Ile Leu Asp Asn Leu Ala Ala Arg Asp Phe 100 105 110 Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr Asp 115 120 <210> 38 <211> 174 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV-GLP-2A2G <400> 38 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln His Gly Asp 130 135 140 Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn Leu Ala Ala 145 150 155 160 Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr Asp 165 170 <210> 39 <211> 186 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV-GLP-2A2G <400> 39 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln 180 185 <210> 40 <211> 241 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-GLP-2A2G <400> 40 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn 210 215 220 Leu Ala Ala Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr 225 230 235 240 Asp <210> 41 <211> 269 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV-GLP-2A2G <400> 41 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln His Gly Asp Gly 225 230 235 240 Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn Leu Ala Ala Arg 245 250 255 Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr Asp 260 265 <210> 42 <211> 326 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV-GLP-2A2G <400> 42 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn 290 295 300 Thr Ile Leu Asp Asn Leu Ala Ala Arg Asp Phe Ile Asn Trp Leu Ile 305 310 315 320 Gln Thr Lys Ile Thr Asp 325 <210> 43 <211> 143 <212> PRT <213> Artificial Sequence <220> <223> SUMO-H6TEV-GLP-2A2G <400> 43 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly His His His His His His Glu Asn Leu Tyr Phe Gln His Gly 100 105 110 Asp Gly Ser Phe Ser Asp Glu Met Asn Thr Ile Leu Asp Asn Leu Ala 115 120 125 Ala Arg Asp Phe Ile Asn Trp Leu Ile Gln Thr Lys Ile Thr Asp 130 135 140 <210> 44 <211> 165 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-INF-beta1b <400> 44 Ser Tyr Asn Leu Leu Gly Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser 1 5 10 15 Gln Lys Leu Leu Trp Gln Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys 20 25 30 Asp Arg Met Asn Phe Asp Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln 35 40 45 Phe Gln Lys Glu Asp Ala Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn 50 55 60 Ile Phe Ala Ile Phe Arg Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu 65 70 75 80 Thr Ile Val Glu Asn Leu Leu Ala Asn Val Tyr His Gln Ile Asn His 85 90 95 Leu Lys Thr Val Leu Glu Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg 100 105 110 Gly Lys Leu Met Ser Ser Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile 115 120 125 Leu His Tyr Leu Lys Ala Lys Glu Tyr Ser His Cys Ala Trp Thr Ile 130 135 140 Val Arg Val Glu Ile Leu Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr 145 150 155 160 Gly Tyr Leu Arg Asn 165 <210> 45 <211> 178 <212> PRT <213> Artificial Sequence <220> <223> H6TEV-INF-beta1b <400> 45 Met His His His His His His Glu Asn Leu Tyr Phe Gln Ser Tyr Asn 1 5 10 15 Leu Leu Gly Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser Gln Lys Leu 20 25 30 Leu Trp Gln Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys Asp Arg Met 35 40 45 Asn Phe Asp Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln Phe Gln Lys 50 55 60 Glu Asp Ala Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn Ile Phe Ala 65 70 75 80 Ile Phe Arg Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu Thr Ile Val 85 90 95 Glu Asn Leu Leu Ala Asn Val Tyr His Gln Ile Asn His Leu Lys Thr 100 105 110 Val Leu Glu Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg Gly Lys Leu 115 120 125 Met Ser Ser Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile Leu His Tyr 130 135 140 Leu Lys Ala Lys Glu Tyr Ser His Cys Ala Trp Thr Ile Val Arg Val 145 150 155 160 Glu Ile Leu Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr Gly Tyr Leu 165 170 175 Arg Asn <210> 46 <211> 255 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV-INF-beta1b <400> 46 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln Ser Tyr Asn Leu Leu Gly 85 90 95 Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser Gln Lys Leu Leu Trp Gln 100 105 110 Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys Asp Arg Met Asn Phe Asp 115 120 125 Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln Phe Gln Lys Glu Asp Ala 130 135 140 Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn Ile Phe Ala Ile Phe Arg 145 150 155 160 Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu Thr Ile Val Glu Asn Leu 165 170 175 Leu Ala Asn Val Tyr His Gln Ile Asn His Leu Lys Thr Val Leu Glu 180 185 190 Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg Gly Lys Leu Met Ser Ser 195 200 205 Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile Leu His Tyr Leu Lys Ala 210 215 220 Lys Glu Tyr Ser His Cys Ala Trp Thr Ile Val Arg Val Glu Ile Leu 225 230 235 240 Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr Gly Tyr Leu Arg Asn 245 250 255 <210> 47 <211> 306 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV-INF-beta1b <400> 47 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln Ser Tyr Asn 130 135 140 Leu Leu Gly Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser Gln Lys Leu 145 150 155 160 Leu Trp Gln Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys Asp Arg Met 165 170 175 Asn Phe Asp Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln Phe Gln Lys 180 185 190 Glu Asp Ala Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn Ile Phe Ala 195 200 205 Ile Phe Arg Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu Thr Ile Val 210 215 220 Glu Asn Leu Leu Ala Asn Val Tyr His Gln Ile Asn His Leu Lys Thr 225 230 235 240 Val Leu Glu Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg Gly Lys Leu 245 250 255 Met Ser Ser Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile Leu His Tyr 260 265 270 Leu Lys Ala Lys Glu Tyr Ser His Cys Ala Trp Thr Ile Val Arg Val 275 280 285 Glu Ile Leu Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr Gly Tyr Leu 290 295 300 Arg Asn 305 <210> 48 <211> 351 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV-INF-beta1b <400> 48 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln Ser Tyr Asn Leu Leu Gly 180 185 190 Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser Gln Lys Leu Leu Trp Gln 195 200 205 Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys Asp Arg Met Asn Phe Asp 210 215 220 Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln Phe Gln Lys Glu Asp Ala 225 230 235 240 Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn Ile Phe Ala Ile Phe Arg 245 250 255 Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu Thr Ile Val Glu Asn Leu 260 265 270 Leu Ala Asn Val Tyr His Gln Ile Asn His Leu Lys Thr Val Leu Glu 275 280 285 Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg Gly Lys Leu Met Ser Ser 290 295 300 Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile Leu His Tyr Leu Lys Ala 305 310 315 320 Lys Glu Tyr Ser His Cys Ala Trp Thr Ile Val Arg Val Glu Ile Leu 325 330 335 Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr Gly Tyr Leu Arg Asn 340 345 350 <210> 49 <211> 373 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-INF-beta1b <400> 49 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 Ser Tyr Asn Leu Leu Gly Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser 210 215 220 Gln Lys Leu Leu Trp Gln Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys 225 230 235 240 Asp Arg Met Asn Phe Asp Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln 245 250 255 Phe Gln Lys Glu Asp Ala Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn 260 265 270 Ile Phe Ala Ile Phe Arg Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu 275 280 285 Thr Ile Val Glu Asn Leu Leu Ala Asn Val Tyr His Gln Ile Asn His 290 295 300 Leu Lys Thr Val Leu Glu Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg 305 310 315 320 Gly Lys Leu Met Ser Ser Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile 325 330 335 Leu His Tyr Leu Lys Ala Lys Glu Tyr Ser His Cys Ala Trp Thr Ile 340 345 350 Val Arg Val Glu Ile Leu Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr 355 360 365 Gly Tyr Leu Arg Asn 370 <210> 50 <211> 401 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV-INF-beta1b <400> 50 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln Ser Tyr Asn Leu 225 230 235 240 Leu Gly Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser Gln Lys Leu Leu 245 250 255 Trp Gln Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys Asp Arg Met Asn 260 265 270 Phe Asp Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln Phe Gln Lys Glu 275 280 285 Asp Ala Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn Ile Phe Ala Ile 290 295 300 Phe Arg Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu Thr Ile Val Glu 305 310 315 320 Asn Leu Leu Ala Asn Val Tyr His Gln Ile Asn His Leu Lys Thr Val 325 330 335 Leu Glu Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg Gly Lys Leu Met 340 345 350 Ser Ser Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile Leu His Tyr Leu 355 360 365 Lys Ala Lys Glu Tyr Ser His Cys Ala Trp Thr Ile Val Arg Val Glu 370 375 380 Ile Leu Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr Gly Tyr Leu Arg 385 390 395 400 Asn <210> 51 <211> 458 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV-INF-beta1b <400> 51 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln Ser Tyr Asn Leu Leu Gly Phe Leu Gln Arg Ser 290 295 300 Ser Asn Phe Gln Ser Gln Lys Leu Leu Trp Gln Leu Asn Gly Arg Leu 305 310 315 320 Glu Tyr Cys Leu Lys Asp Arg Met Asn Phe Asp Ile Pro Glu Glu Ile 325 330 335 Lys Gln Leu Gln Gln Phe Gln Lys Glu Asp Ala Ala Leu Thr Ile Tyr 340 345 350 Glu Met Leu Gln Asn Ile Phe Ala Ile Phe Arg Gln Asp Ser Ser Ser 355 360 365 Thr Gly Trp Asn Glu Thr Ile Val Glu Asn Leu Leu Ala Asn Val Tyr 370 375 380 His Gln Ile Asn His Leu Lys Thr Val Leu Glu Glu Lys Leu Glu Lys 385 390 395 400 Glu Asp Phe Thr Arg Gly Lys Leu Met Ser Ser Leu His Leu Lys Arg 405 410 415 Tyr Tyr Gly Arg Ile Leu His Tyr Leu Lys Ala Lys Glu Tyr Ser His 420 425 430 Cys Ala Trp Thr Ile Val Arg Val Glu Ile Leu Arg Asn Phe Tyr Phe 435 440 445 Ile Asn Arg Leu Thr Gly Tyr Leu Arg Asn 450 455 <210> 52 <211> 275 <212> PRT <213> Artificial Sequence <220> <223> SUMO-H6TEV-INF-beta1b <400> 52 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly His His His His His His Glu Asn Leu Tyr Phe Gln Ser Tyr 100 105 110 Asn Leu Leu Gly Phe Leu Gln Arg Ser Ser Asn Phe Gln Ser Gln Lys 115 120 125 Leu Leu Trp Gln Leu Asn Gly Arg Leu Glu Tyr Cys Leu Lys Asp Arg 130 135 140 Met Asn Phe Asp Ile Pro Glu Glu Ile Lys Gln Leu Gln Gln Phe Gln 145 150 155 160 Lys Glu Asp Ala Ala Leu Thr Ile Tyr Glu Met Leu Gln Asn Ile Phe 165 170 175 Ala Ile Phe Arg Gln Asp Ser Ser Ser Thr Gly Trp Asn Glu Thr Ile 180 185 190 Val Glu Asn Leu Leu Ala Asn Val Tyr His Gln Ile Asn His Leu Lys 195 200 205 Thr Val Leu Glu Glu Lys Leu Glu Lys Glu Asp Phe Thr Arg Gly Lys 210 215 220 Leu Met Ser Ser Leu His Leu Lys Arg Tyr Tyr Gly Arg Ile Leu His 225 230 235 240 Tyr Leu Lys Ala Lys Glu Tyr Ser His Cys Ala Trp Thr Ile Val Arg 245 250 255 Val Glu Ile Leu Arg Asn Phe Tyr Phe Ile Asn Arg Leu Thr Gly Tyr 260 265 270 Leu Arg Asn 275 <210> 53 <211> 86 <212> PRT <213> Artificial Sequence <220> <223> Insulin glargine precursor (INSG) <400> 53 Phe Val Asn Gln His Leu Cys Gly Ser His Leu Val Glu Ala Leu Tyr 1 5 10 15 Leu Val Cys Gly Glu Arg Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg 20 25 30 Glu Ala Glu Asp Leu Gln Val Gly Gln Val Glu Leu Gly Gly Gly Pro 35 40 45 Gly Ala Gly Ser Leu Gln Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys 50 55 60 Arg Gly Ile Val Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln 65 70 75 80 Leu Glu Asn Tyr Cys Gly 85 <210> 54 <211> 99 <212> PRT <213> Artificial Sequence <220> <223> H6TEV-INSG <400> 54 Met His His His His His His Glu Asn Leu Tyr Phe Gln Phe Val Asn 1 5 10 15 Gln His Leu Cys Gly Ser His Leu Val Glu Ala Leu Tyr Leu Val Cys 20 25 30 Gly Glu Arg Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala Glu 35 40 45 Asp Leu Gln Val Gly Gln Val Glu Leu Gly Gly Gly Pro Gly Ala Gly 50 55 60 Ser Leu Gln Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly Ile 65 70 75 80 Val Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu Asn 85 90 95 Tyr Cys Gly <210> 55 <211> 176 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV-INSG <400> 55 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln Phe Val Asn Gln His Leu 85 90 95 Cys Gly Ser His Leu Val Glu Ala Leu Tyr Leu Val Cys Gly Glu Arg 100 105 110 Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala Glu Asp Leu Gln 115 120 125 Val Gly Gln Val Glu Leu Gly Gly Gly Pro Gly Ala Gly Ser Leu Gln 130 135 140 Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly Ile Val Glu Gln 145 150 155 160 Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu Asn Tyr Cys Gly 165 170 175 <210> 56 <211> 227 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV-INSG <400> 56 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln Phe Val Asn 130 135 140 Gln His Leu Cys Gly Ser His Leu Val Glu Ala Leu Tyr Leu Val Cys 145 150 155 160 Gly Glu Arg Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala Glu 165 170 175 Asp Leu Gln Val Gly Gln Val Glu Leu Gly Gly Gly Pro Gly Ala Gly 180 185 190 Ser Leu Gln Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly Ile 195 200 205 Val Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu Asn 210 215 220 Tyr Cys Gly 225 <210> 57 <211> 272 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV-INSG <400> 57 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln Phe Val Asn Gln His Leu 180 185 190 Cys Gly Ser His Leu Val Glu Ala Leu Tyr Leu Val Cys Gly Glu Arg 195 200 205 Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala Glu Asp Leu Gln 210 215 220 Val Gly Gln Val Glu Leu Gly Gly Gly Pro Gly Ala Gly Ser Leu Gln 225 230 235 240 Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly Ile Val Glu Gln 245 250 255 Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu Asn Tyr Cys Gly 260 265 270 <210> 58 <211> 294 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-INSG <400> 58 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 Phe Val Asn Gln His Leu Cys Gly Ser His Leu Val Glu Ala Leu Tyr 210 215 220 Leu Val Cys Gly Glu Arg Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg 225 230 235 240 Glu Ala Glu Asp Leu Gln Val Gly Gln Val Glu Leu Gly Gly Gly Pro 245 250 255 Gly Ala Gly Ser Leu Gln Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys 260 265 270 Arg Gly Ile Val Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln 275 280 285 Leu Glu Asn Tyr Cys Gly 290 <210> 59 <211> 322 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV-INSG <400> 59 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln Phe Val Asn Gln 225 230 235 240 His Leu Cys Gly Ser His Leu Val Glu Ala Leu Tyr Leu Val Cys Gly 245 250 255 Glu Arg Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala Glu Asp 260 265 270 Leu Gln Val Gly Gln Val Glu Leu Gly Gly Gly Pro Gly Ala Gly Ser 275 280 285 Leu Gln Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly Ile Val 290 295 300 Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu Asn Tyr 305 310 315 320 Cys Gly <210> 60 <211> 379 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV-INSG <400> 60 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln Phe Val Asn Gln His Leu Cys Gly Ser His Leu 290 295 300 Val Glu Ala Leu Tyr Leu Val Cys Gly Glu Arg Gly Phe Phe Tyr Thr 305 310 315 320 Pro Lys Thr Arg Arg Glu Ala Glu Asp Leu Gln Val Gly Gln Val Glu 325 330 335 Leu Gly Gly Gly Pro Gly Ala Gly Ser Leu Gln Pro Leu Ala Leu Glu 340 345 350 Gly Ser Leu Gln Lys Arg Gly Ile Val Glu Gln Cys Cys Thr Ser Ile 355 360 365 Cys Ser Leu Tyr Gln Leu Glu Asn Tyr Cys Gly 370 375 <210> 61 <211> 196 <212> PRT <213> Artificial Sequence <220> <223> SUMO-H6TEV-INSG <400> 61 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly His His His His His His Glu Asn Leu Tyr Phe Gln Phe Val 100 105 110 Asn Gln His Leu Cys Gly Ser His Leu Val Glu Ala Leu Tyr Leu Val 115 120 125 Cys Gly Glu Arg Gly Phe Phe Tyr Thr Pro Lys Thr Arg Arg Glu Ala 130 135 140 Glu Asp Leu Gln Val Gly Gln Val Glu Leu Gly Gly Gly Pro Gly Ala 145 150 155 160 Gly Ser Leu Gln Pro Leu Ala Leu Glu Gly Ser Leu Gln Lys Arg Gly 165 170 175 Ile Val Glu Gln Cys Cys Thr Ser Ile Cys Ser Leu Tyr Gln Leu Glu 180 185 190 Asn Tyr Cys Gly 195 <210> 62 <211> 191 <212> PRT <213> Artificial Sequence <220> <223> Human growth hormon (hGH) <400> 62 Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 1 5 10 15 Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 20 25 30 Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 35 40 45 Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 50 55 60 Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 65 70 75 80 Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 85 90 95 Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 100 105 110 Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 115 120 125 Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 130 135 140 Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 145 150 155 160 Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 165 170 175 Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 180 185 190 <210> 63 <211> 204 <212> PRT <213> Artificial Sequence <220> <223> H6TEV-hGH <400> 63 Met His His His His His His Glu Asn Leu Tyr Phe Gln Phe Pro Thr 1 5 10 15 Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg 20 25 30 Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr 35 40 45 Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser 50 55 60 Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr 65 70 75 80 Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile 85 90 95 Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn 100 105 110 Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys 115 120 125 Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly 130 135 140 Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp 145 150 155 160 Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu 165 170 175 Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile 180 185 190 Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 195 200 <210> 64 <211> 281 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV-hGH <400> 64 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln Phe Pro Thr Ile Pro Leu 85 90 95 Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln 100 105 110 Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys 115 120 125 Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe 130 135 140 Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys 145 150 155 160 Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp 165 170 175 Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val 180 185 190 Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu 195 200 205 Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg 210 215 220 Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser 225 230 235 240 His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe 245 250 255 Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys 260 265 270 Arg Ser Val Glu Gly Ser Cys Gly Phe 275 280 <210> 65 <211> 332 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV-hGH <400> 65 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln Phe Pro Thr 130 135 140 Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg 145 150 155 160 Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr 165 170 175 Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser 180 185 190 Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr 195 200 205 Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile 210 215 220 Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn 225 230 235 240 Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys 245 250 255 Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly 260 265 270 Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp 275 280 285 Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu 290 295 300 Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile 305 310 315 320 Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 325 330 <210> 66 <211> 377 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV-hGH <400> 66 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln Phe Pro Thr Ile Pro Leu 180 185 190 Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu His Gln 195 200 205 Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys 210 215 220 Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe 225 230 235 240 Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys 245 250 255 Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp 260 265 270 Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val 275 280 285 Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu 290 295 300 Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg 305 310 315 320 Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser 325 330 335 His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe 340 345 350 Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys 355 360 365 Arg Ser Val Glu Gly Ser Cys Gly Phe 370 375 <210> 67 <211> 399 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-hGH <400> 67 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg 210 215 220 Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu 225 230 235 240 Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro 245 250 255 Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg 260 265 270 Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu 275 280 285 Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val 290 295 300 Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp 305 310 315 320 Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu 325 330 335 Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser 340 345 350 Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr 355 360 365 Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe 370 375 380 Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 385 390 395 <210> 68 <211> 427 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV-hGH <400> 68 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln Phe Pro Thr Ile 225 230 235 240 Pro Leu Ser Arg Leu Phe Asp Asn Ala Met Leu Arg Ala His Arg Leu 245 250 255 His Gln Leu Ala Phe Asp Thr Tyr Gln Glu Phe Glu Glu Ala Tyr Ile 260 265 270 Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln Asn Pro Gln Thr Ser Leu 275 280 285 Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser Asn Arg Glu Glu Thr Gln 290 295 300 Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile Ser Leu Leu Leu Ile Gln 305 310 315 320 Ser Trp Leu Glu Pro Val Gln Phe Leu Arg Ser Val Phe Ala Asn Ser 325 330 335 Leu Val Tyr Gly Ala Ser Asp Ser Asn Val Tyr Asp Leu Leu Lys Asp 340 345 350 Leu Glu Glu Gly Ile Gln Thr Leu Met Gly Arg Leu Glu Asp Gly Ser 355 360 365 Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr Tyr Ser Lys Phe Asp Thr 370 375 380 Asn Ser His Asn Asp Asp Ala Leu Leu Lys Asn Tyr Gly Leu Leu Tyr 385 390 395 400 Cys Phe Arg Lys Asp Met Asp Lys Val Glu Thr Phe Leu Arg Ile Val 405 410 415 Gln Cys Arg Ser Val Glu Gly Ser Cys Gly Phe 420 425 <210> 69 <211> 484 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV-hGH <400> 69 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp 290 295 300 Asn Ala Met Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr 305 310 315 320 Tyr Gln Glu Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser 325 330 335 Phe Leu Gln Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro 340 345 350 Thr Pro Ser Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu 355 360 365 Leu Arg Ile Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln 370 375 380 Phe Leu Arg Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp 385 390 395 400 Ser Asn Val Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr 405 410 415 Leu Met Gly Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe 420 425 430 Lys Gln Thr Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala 435 440 445 Leu Leu Lys Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp 450 455 460 Lys Val Glu Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly 465 470 475 480 Ser Cys Gly Phe <210> 70 <211> 305 <212> PRT <213> Artificial Sequence <220> <223> SUMO-H6TEV-hGH <400> 70 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly His His His His His His Glu Asn Leu Tyr Phe Gln Phe Val 100 105 110 Asn Gln Phe Pro Thr Ile Pro Leu Ser Arg Leu Phe Asp Asn Ala Met 115 120 125 Leu Arg Ala His Arg Leu His Gln Leu Ala Phe Asp Thr Tyr Gln Glu 130 135 140 Phe Glu Glu Ala Tyr Ile Pro Lys Glu Gln Lys Tyr Ser Phe Leu Gln 145 150 155 160 Asn Pro Gln Thr Ser Leu Cys Phe Ser Glu Ser Ile Pro Thr Pro Ser 165 170 175 Asn Arg Glu Glu Thr Gln Gln Lys Ser Asn Leu Glu Leu Leu Arg Ile 180 185 190 Ser Leu Leu Leu Ile Gln Ser Trp Leu Glu Pro Val Gln Phe Leu Arg 195 200 205 Ser Val Phe Ala Asn Ser Leu Val Tyr Gly Ala Ser Asp Ser Asn Val 210 215 220 Tyr Asp Leu Leu Lys Asp Leu Glu Glu Gly Ile Gln Thr Leu Met Gly 225 230 235 240 Arg Leu Glu Asp Gly Ser Pro Arg Thr Gly Gln Ile Phe Lys Gln Thr 245 250 255 Tyr Ser Lys Phe Asp Thr Asn Ser His Asn Asp Asp Ala Leu Leu Lys 260 265 270 Asn Tyr Gly Leu Leu Tyr Cys Phe Arg Lys Asp Met Asp Lys Val Glu 275 280 285 Thr Phe Leu Arg Ile Val Gln Cys Arg Ser Val Glu Gly Ser Cys Gly 290 295 300 Phe 305 <210> 71 <211> 37 <212> PRT <213> Artificial Sequence <220> <223> Oxyntomodulin (OXM) <400> 71 His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser 1 5 10 15 Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr Lys Arg Asn 20 25 30 Arg Asn Asn Ile Ala 35 <210> 72 <211> 50 <212> PRT <213> Artificial Sequence <220> <223> H6TEV-OXM <400> 72 Met His His His His His His Glu Asn Leu Tyr Phe Gln His Ser Gln 1 5 10 15 Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser Arg Arg Ala 20 25 30 Gln Asp Phe Val Gln Trp Leu Met Asn Thr Lys Arg Asn Arg Asn Asn 35 40 45 Ile Ala 50 <210> 73 <211> 127 <212> PRT <213> Artificial Sequence <220> <223> IMC-H6TEV-OXM <400> 73 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr His His 65 70 75 80 His His His His Glu Asn Leu Tyr Phe Gln His Ser Gln Gly Thr Phe 85 90 95 Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser Arg Arg Ala Gln Asp Phe 100 105 110 Val Gln Trp Leu Met Asn Thr Lys Arg Asn Arg Asn Asn Ile Ala 115 120 125 <210> 74 <211> 178 <212> PRT <213> Artificial Sequence <220> <223> IMCB1-H6TEV-OXM <400> 74 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val His His His His His His Glu Asn Leu Tyr Phe Gln His Ser Gln 130 135 140 Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser Arg Arg Ala 145 150 155 160 Gln Asp Phe Val Gln Trp Leu Met Asn Thr Lys Arg Asn Arg Asn Asn 165 170 175 Ile Ala <210> 75 <211> 186 <212> PRT <213> Artificial Sequence <220> <223> IMCB2-H6TEV-OXM <400> 75 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu His His 165 170 175 His His His His Glu Asn Leu Tyr Phe Gln 180 185 <210> 76 <211> 245 <212> PRT <213> Artificial Sequence <220> <223> IMCC1-H6TEV-OXM <400> 76 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn His His His His His His Glu Asn Leu Tyr Phe Gln 195 200 205 His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser 210 215 220 Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr Lys Arg Asn 225 230 235 240 Arg Asn Asn Ile Ala 245 <210> 77 <211> 273 <212> PRT <213> Artificial Sequence <220> <223> IMCC2-H6TEV-OXM <400> 77 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 His His His His His His Glu Asn Leu Tyr Phe Gln His Ser Gln Gly 225 230 235 240 Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser Arg Arg Ala Gln 245 250 255 Asp Phe Val Gln Trp Leu Met Asn Thr Lys Arg Asn Arg Asn Asn Ile 260 265 270 Ala <210> 78 <211> 330 <212> PRT <213> Artificial Sequence <220> <223> IMCD1-H6TEV-OXM <400> 78 Met Ala Gly Lys Ser Ser Thr Glu Lys Lys Tyr Ile Val Gly Phe Lys 1 5 10 15 Gln Thr Met Ser Ala Met Ser Ser Ala Lys Lys Lys Asp Val Ile Ser 20 25 30 Glu Lys Gly Gly Lys Val Gln Lys Gln Phe Lys Tyr Val Asn Ala Ala 35 40 45 Ala Ala Thr Leu Asp Glu Lys Ala Val Lys Glu Leu Lys Lys Asp Pro 50 55 60 Ser Val Ala Tyr Val Glu Glu Asp His Ile Ala His Glu Tyr Ala Gln 65 70 75 80 Ser Phe Pro Tyr Gly Ile Ser Gln Ile Lys Ala Pro Ala Leu His Ser 85 90 95 Gln Gly Tyr Thr Gly Ser Asn Val Lys Val Ala Val Ile Asp Ser Gly 100 105 110 Ile Asp Ser Ser His Pro Asp Leu Asn Val Arg Gly Gly Ala Ser Phe 115 120 125 Val Pro Ser Glu Thr Asn Pro Tyr Gln Asp Gly Ser Ser His Gly Thr 130 135 140 His Val Ala Gly Thr Ile Ala Ala Leu Asn Asn Ser Ile Gly Val Leu 145 150 155 160 Gly Val Ala Pro Ser Ala Ser Leu Tyr Ala Val Lys Val Leu Asp Ser 165 170 175 Thr Gly Ser Gly Gln Tyr Ser Trp Ile Ile Asn Gly Ile Glu Trp Ala 180 185 190 Ile Ser Asn Asn Met Asp Val Ile Asn Met Ser Leu Gly Gly Pro Thr 195 200 205 Gly Ser Thr Ala Leu Lys Thr Val Val Asp Lys Ala Val Ser Ser Gly 210 215 220 Ile Val Val Ala Ala Ala Ala Gly Asn Glu Gly Ser Ser Gly Ser Thr 225 230 235 240 Ser Thr Val Gly Tyr Pro Ala Lys Tyr Pro Ser Thr Ile Ala Val Gly 245 250 255 Ala Val Asn Ser Ser Asn Gln Arg Ala Ser Phe Ser Ser Ala Gly Ser 260 265 270 Glu Leu Asp Val Met Ala Pro Gly Val His His His His His His Glu 275 280 285 Asn Leu Tyr Phe Gln His Ser Gln Gly Thr Phe Thr Ser Asp Tyr Ser 290 295 300 Lys Tyr Leu Asp Ser Arg Arg Ala Gln Asp Phe Val Gln Trp Leu Met 305 310 315 320 Asn Thr Lys Arg Asn Arg Asn Asn Ile Ala 325 330 <210> 79 <211> 147 <212> PRT <213> Artificial Sequence <220> <223> SUMO-H6TEV-OXM <400> 79 Met Ser Asp Ser Glu Val Asn Gln Glu Ala Lys Pro Glu Val Lys Pro 1 5 10 15 Glu Val Lys Pro Glu Thr His Ile Asn Leu Lys Val Ser Asp Gly Ser 20 25 30 Ser Glu Ile Phe Phe Lys Ile Lys Lys Thr Thr Pro Leu Arg Arg Leu 35 40 45 Met Glu Ala Phe Ala Lys Arg Gln Gly Lys Glu Met Asp Ser Leu Arg 50 55 60 Phe Leu Tyr Asp Gly Ile Arg Ile Gln Ala Asp Gln Thr Pro Glu Asp 65 70 75 80 Leu Asp Met Glu Asp Asn Asp Ile Ile Glu Ala His Arg Glu Gln Ile 85 90 95 Gly Gly His His His His His His Glu Asn Leu Tyr Phe Gln His Ser 100 105 110 Gln Gly Thr Phe Thr Ser Asp Tyr Ser Lys Tyr Leu Asp Ser Arg Arg 115 120 125 Ala Gln Asp Phe Val Gln Trp Leu Met Asn Thr Lys Arg Asn Arg Asn 130 135 140 Asn Ile Ala 145 <210> 80 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer (lacI-5UTR-F) <400> 80 cgtgactggg tcatggctgc g 21 <210> 81 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer (rrnB-term-IF-R) <400> 81 aaggcccagt ctttcgactg a 21 <210> 82 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer (rrnB-term-IF-F) <400> 82 tcagtcgaaa gactgggcct t 21 <210> 83 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer (lacI-5UTR-R) <400> 83 cgcagccatg acccagtcac g 21 <210> 84 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> Primer (pSG01-pelB-IF-F) <400> 84 aataaggagg aataacatat gaaatacctg ctgccg 36 <210> 85 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> Primer (6his-pSG01-IF-R) <400> 85 gaccgtttaa actcagtggt ggtggtggtg gtgctc 36 <210> 86 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer (pSK01-IF-F) <400> 86 tgagtttaaa cggtctccag c 21 <210> 87 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer (pSK01-IF-R) <400> 87 cgcagccatg acccagtcac g 21 <210> 88 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> Primer (T7ter-rrnBter-F) <400> 88 gtcttgaggg gttttttgag cggtctgata aaacag 36 <210> 89 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> Primer (lacO-R) <400> 89 gggaattgtt atccgctcac aattcc 26 <210> 90 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> Primer (lacO-F) <400> 90 ggaattgtga gcggataaca attccc 26 <210> 91 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> Primer (T7ter-rrnBter-R) <400> 91 ctgttttatc agaccgctca aaaaacccct caagac 36 <210> 92 <211> 237 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for N-terminal fusion partner (IMC) <400> 92 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atattaa 237 <210> 93 <211> 390 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for N-terminal fusion partner (IMC-Fragment B1 (IMCB1)) <400> 93 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtataa 390 <210> 94 <211> 525 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for N-terminal fusion partner (IMC-Fragment B2 (IMCB2)) <400> 94 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc tttaa 525 <210> 95 <211> 591 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for N-terminal fusion partner (IMC-Fragment C1 (IMCC1)) <400> 95 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatta a 591 <210> 96 <211> 675 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for N-terminal fusion partner (IMC-Fragment C2 (IMCC2)) <400> 96 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gttaa 675 <210> 97 <211> 846 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for N-terminal fusion partner (IMC-Fragment D1 (IMCD1)) <400> 97 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgtaa 846 <210> 98 <211> 294 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO <400> 98 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggt 294 <210> 99 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for Histidine 6 <400> 99 catcatcatc accaccac 18 <210> 100 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for TEV protease recognition sequence <400> 100 gaaaacctgt atttccag 18 <210> 101 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV <400> 101 catcatcatc accaccacga aaacctgtat ttccag 36 <210> 102 <211> 273 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV <400> 102 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag taa 273 <210> 103 <211> 426 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV <400> 103 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagtaa 426 <210> 104 <211> 561 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV <400> 104 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagta a 561 <210> 105 <211> 627 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV <400> 105 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagtaa 627 <210> 106 <211> 711 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV <400> 106 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagta a 711 <210> 107 <211> 882 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV <400> 107 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgcatcatc atcaccacca cgaaaacctg tatttccagt aa 882 <210> 108 <211> 96 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for Liraglutide precursor peptide (GLP-1K28R) <400> 108 catgcggaag gcacctttac cagcgatgtg agcagctatc tggaaggcca ggcggcgaaa 60 gaatttattg cgtggctggt gcgtggccgc ggctaa 96 <210> 109 <211> 135 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV-GLP-1K28R <400> 109 atgcatcatc atcaccacca cgaaaacctg tatttccagc atgcggaagg cacctttacc 60 agcgatgtga gcagctatct ggaaggccag gcggcgaaag aatttattgc gtggctggtg 120 cgtggccgcg gctaa 135 <210> 110 <211> 366 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV-GLP-1K28R <400> 110 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag catgcggaag gcacctttac cagcgatgtg 300 agcagctatc tggaaggcca ggcggcgaaa gaatttattg cgtggctggt gcgtggccgc 360 ggctaa 366 <210> 111 <211> 522 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV-GLP-1K28R <400> 111 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagtaacatg cggaaggcac ctttaccagc gatgtgagca gctatctgga aggccaggcg 480 gcgaaagaat ttattgcgtg gctggtgcgt ggccgcggct aa 522 <210> 112 <211> 561 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV-GLP-1K28R <400> 112 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagta a 561 <210> 113 <211> 627 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV-GLP-1K28R <400> 113 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagtaa 627 <210> 114 <211> 711 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV-GLP-1K28R <400> 114 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagta a 711 <210> 115 <211> 882 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV-GLP-1K28R <400> 115 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgcatcatc atcaccacca cgaaaacctg tatttccagt aa 882 <210> 116 <211> 426 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO-H6TEV-GLP-1K28R <400> 116 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag catgcggaag gcacctttac cagcgatgtg 360 agcagctatc tggaaggcca ggcggcgaaa gaatttattg cgtggctggt gcgtggccgc 420 ggctaa 426 <210> 117 <211> 105 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for hPTH1-34 (Teriparatide) <400> 117 agcgtgagcg aaattcagct gatgcataac ctgggcaaac atctgaacag catggaacgc 60 gtggaatggc tgcgcaaaaa actgcaggat gtgcataact tttaa 105 <210> 118 <211> 144 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV-hPTH1-34 <400> 118 atgcatcatc atcaccacca cgaaaacctg tatttccaga gcgtgagcga aattcagctg 60 atgcataacc tgggcaaaca tctgaacagc atggaacgcg tggaatggct gcgcaaaaaa 120 ctgcaggatg tgcataactt ttaa 144 <210> 119 <211> 375 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV-hPTH1-34 <400> 119 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag agcgtgagcg aaattcagct gatgcataac 300 ctgggcaaac atctgaacag catggaacgc gtggaatggc tgcgcaaaaa actgcaggat 360 gtgcataact tttaa 375 <210> 120 <211> 528 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV-hPTH1-34 <400> 120 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagagcgtga gcgaaattca gctgatgcat aacctgggca aacatctgaa cagcatggaa 480 cgcgtggaat ggctgcgcaa aaaactgcag gatgtgcata acttttaa 528 <210> 121 <211> 663 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV-hPTH1-34 <400> 121 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagag cgtgagcgaa attcagctga tgcataacct gggcaaacat 600 ctgaacagca tggaacgcgt ggaatggctg cgcaaaaaac tgcaggatgt gcataacttt 660 taa 663 <210> 122 <211> 729 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV-hPTH1-34 <400> 122 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagagcgtg agcgaaattc agctgatgca taacctgggc 660 aaacatctga acagcatgga acgcgtggaa tggctgcgca aaaaactgca ggatgtgcat 720 aacttttaa 729 <210> 123 <211> 813 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV-hPTH1-34 <400> 123 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagag cgtgagcgaa 720 attcagctga tgcataacct gggcaaacat ctgaacagca tggaacgcgt ggaatggctg 780 cgcaaaaaac tgcaggatgt gcataacttt taa 813 <210> 124 <211> 531 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV-hPTH1-34 <400> 124 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag catgcggaag gcacctttac cagcgatgtg 360 agcagctatc tggaaggcca ggcggcgaaa gaatttattg cgtggctggt gcgtggccgc 420 ggctaaagcg tgagcgaaat tcagctgatg cataacctgg gcaaacatct gaacagcatg 480 gaacgcgtgg aatggctgcg caaaaaactg caggatgtgc ataactttta a 531 <210> 125 <211> 435 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO-H6TEV-hPTH1-34 <400> 125 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag agcgtgagcg aaattcagct gatgcataac 360 ctgggcaaac atctgaacag catggaacgc gtggaatggc tgcgcaaaaa actgcaggat 420 gtgcataact tttaa 435 <210> 126 <211> 102 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for Teduglutide (GLP-2A2G) <400> 126 catggcgatg gcagctttag cgatgaaatg aacaccattc tggataacct ggcggcgcgc 60 gattttatta actggctgat tcagaccaaa attaccgatt aa 102 <210> 127 <211> 141 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV-GLP-2A2G <400> 127 atgcatcatc atcaccacca cgaaaacctg tatttccagc atggcgatgg cagctttagc 60 gatgaaatga acaccattct ggataacctg gcggcgcgcg attttattaa ctggctgatt 120 cagaccaaaa ttaccgatta a 141 <210> 128 <211> 372 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV-GLP-2A2G <400> 128 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag catggcgatg gcagctttag cgatgaaatg 300 aacaccattc tggataacct ggcggcgcgc gattttatta actggctgat tcagaccaaa 360 attaccgatt aa 372 <210> 129 <211> 525 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV-GLP-2A2G <400> 129 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagcatgcgg atggcagctt tagcgatgaa atgaacacca ttctggataa cctggcggcg 480 cgcgatttta ttaactggct gattcagacc aaaattaccg attaa 525 <210> 130 <211> 660 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV-GLP-2A2G <400> 130 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagca tgcggatggc agctttagcg atgaaatgaa caccattctg 600 gataacctgg cggcgcgcga ttttattaac tggctgattc agaccaaaat taccgattaa 660 660 <210> 131 <211> 726 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV-GLP-2A2G <400> 131 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagcatgcg gatggcagct ttagcgatga aatgaacacc 660 attctggata acctggcggc gcgcgatttt attaactggc tgattcagac caaaattacc 720 gattaa 726 <210> 132 <211> 810 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV-GLP-2A2G <400> 132 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagca tgcggatggc 720 agctttagcg atgaaatgaa caccattctg gataacctgg cggcgcgcga ttttattaac 780 tggctgattc agaccaaaat taccgattaa 810 <210> 133 <211> 981 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV-GLP-2A2G <400> 133 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgcatcatc atcaccacca cgaaaacctg tatttccagc atgcggatgg cagctttagc 900 gatgaaatga acaccattct ggataacctg gcggcgcgcg attttattaa ctggctgatt 960 cagaccaaaa ttaccgatta a 981 <210> 134 <211> 432 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO-H6TEV-GLP-2A2G <400> 134 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag catggcgatg gcagctttag cgatgaaatg 360 aacaccattc tggataacct ggcggcgcgc gattttatta actggctgat tcagaccaaa 420 attaccgatt aa 432 <210> 135 <211> 498 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for Interferon beta1b (INF-beta1b) <400> 135 agctataacc tgctgggctt tctgcagcgc agcagcaact ttcagagcca gaaactgctg 60 tggcagctga acggccgcct ggaatattgc ctgaaagatc gcatgaactt tgatattccg 120 gaagaaatta aacagctgca gcagtttcag aaagaagatg cggcgctgac catttatgaa 180 atgctgcaga acatttttgc gatttttcgc caggatagca gcagcaccgg ctggaacgaa 240 accattgtgg aaaacctgct ggcgaacgtg tatcatcaga ttaaccatct gaaaaccgtg 300 ctggaagaaa aactggaaaa agaagatttt acccgcggca aactgatgag cagcctgcat 360 ctgaaacgct attatggccg cattctgcat tatctgaaag cgaaagaata tagccattgc 420 gcgtggacca ttgtgcgcgt ggaaattctg cgcaactttt attttattaa ccgcctgacc 480 ggctatctgc gcaactaa 498 <210> 136 <211> 537 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV-INF-beta1b <400> 136 atgcatcatc atcaccacca cgaaaacctg tatttccaga gctataacct gctgggcttt 60 ctgcagcgca gcagcaactt tcagagccag aaactgctgt ggcagctgaa cggccgcctg 120 gaatattgcc tgaaagatcg catgaacttt gatattccgg aagaaattaa acagctgcag 180 cagtttcaga aagaagatgc ggcgctgacc atttatgaaa tgctgcagaa catttttgcg 240 atttttcgcc aggatagcag cagcaccggc tggaacgaaa ccattgtgga aaacctgctg 300 gcgaacgtgt atcatcagat taaccatctg aaaaccgtgc tggaagaaaa actggaaaaa 360 gaagatttta cccgcggcaa actgatgagc agcctgcatc tgaaacgcta ttatggccgc 420 attctgcatt atctgaaagc gaaagaatat agccattgcg cgtggaccat tgtgcgcgtg 480 gaaattctgc gcaactttta ttttattaac cgcctgaccg gctatctgcg caactaa 537 <210> 137 <211> 768 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV-INF-beta1b <400> 137 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag agctataacc tgctgggctt tctgcagcgc 300 agcagcaact ttcagagcca gaaactgctg tggcagctga acggccgcct ggaatattgc 360 ctgaaagatc gcatgaactt tgatattccg gaagaaatta aacagctgca gcagtttcag 420 aaagaagatg cggcgctgac catttatgaa atgctgcaga acatttttgc gatttttcgc 480 caggatagca gcagcaccgg ctggaacgaa accattgtgg aaaacctgct ggcgaacgtg 540 tatcatcaga ttaaccatct gaaaaccgtg ctggaagaaa aactggaaaa agaagatttt 600 acccgcggca aactgatgag cagcctgcat ctgaaacgct attatggccg cattctgcat 660 tatctgaaag cgaaagaata tagccattgc gcgtggacca ttgtgcgcgt ggaaattctg 720 cgcaactttt attttattaa ccgcctgacc ggctatctgc gcaactaa 768 <210> 138 <211> 921 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV-INF-beta1b <400> 138 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagagctata acctgctggg ctttctgcag cgcagcagca actttcagag ccagaaactg 480 ctgtggcagc tgaacggccg cctggaatat tgcctgaaag atcgcatgaa ctttgatatt 540 ccggaagaaa ttaaacagct gcagcagttt cagaaagaag atgcggcgct gaccatttat 600 gaaatgctgc agaacatttt tgcgattttt cgccaggata gcagcagcac cggctggaac 660 gaaaccattg tggaaaacct gctggcgaac gtgtatcatc agattaacca tctgaaaacc 720 gtgctggaag aaaaactgga aaaagaagat tttacccgcg gcaaactgat gagcagcctg 780 catctgaaac gctattatgg ccgcattctg cattatctga aagcgaaaga atatagccat 840 tgcgcgtgga ccattgtgcg cgtggaaatt ctgcgcaact tttattttat taaccgcctg 900 accggctatc tgcgcaacta a 921 <210> 139 <211> 1056 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV-INF-beta1b <400> 139 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagag ctataacctg ctgggctttc tgcagcgcag cagcaacttt 600 cagagccaga aactgctgtg gcagctgaac ggccgcctgg aatattgcct gaaagatcgc 660 atgaactttg atattccgga agaaattaaa cagctgcagc agtttcagaa agaagatgcg 720 gcgctgacca tttatgaaat gctgcagaac atttttgcga tttttcgcca ggatagcagc 780 agcaccggct ggaacgaaac cattgtggaa aacctgctgg cgaacgtgta tcatcagatt 840 aaccatctga aaaccgtgct ggaagaaaaa ctggaaaaag aagattttac ccgcggcaaa 900 ctgatgagca gcctgcatct gaaacgctat tatggccgca ttctgcatta tctgaaagcg 960 aaagaatata gccattgcgc gtggaccatt gtgcgcgtgg aaattctgcg caacttttat 1020 tttattaacc gcctgaccgg ctatctgcgc aactaa 1056 <210> 140 <211> 1122 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV-INF-beta1b <400> 140 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagagctat aacctgctgg gctttctgca gcgcagcagc 660 aactttcaga gccagaaact gctgtggcag ctgaacggcc gcctggaata ttgcctgaaa 720 gatcgcatga actttgatat tccggaagaa attaaacagc tgcagcagtt tcagaaagaa 780 gatgcggcgc tgaccattta tgaaatgctg cagaacattt ttgcgatttt tcgccaggat 840 agcagcagca ccggctggaa cgaaaccatt gtggaaaacc tgctggcgaa cgtgtatcat 900 cagattaacc atctgaaaac cgtgctggaa gaaaaactgg aaaaagaaga ttttacccgc 960 ggcaaactga tgagcagcct gcatctgaaa cgctattatg gccgcattct gcattatctg 1020 aaagcgaaag aatatagcca ttgcgcgtgg accattgtgc gcgtggaaat tctgcgcaac 1080 ttttatttta ttaaccgcct gaccggctat ctgcgcaact aa 1122 <210> 141 <211> 1206 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV-INF-beta1b <400> 141 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagag ctataacctg 720 ctgggctttc tgcagcgcag cagcaacttt cagagccaga aactgctgtg gcagctgaac 780 ggccgcctgg aatattgcct gaaagatcgc atgaactttg atattccgga agaaattaaa 840 cagctgcagc agtttcagaa agaagatgcg gcgctgacca tttatgaaat gctgcagaac 900 atttttgcga tttttcgcca ggatagcagc agcaccggct ggaacgaaac cattgtggaa 960 aacctgctgg cgaacgtgta tcatcagatt aaccatctga aaaccgtgct ggaagaaaaa 1020 ctggaaaaag aagattttac ccgcggcaaa ctgatgagca gcctgcatct gaaacgctat 1080 tatggccgca ttctgcatta tctgaaagcg aaagaatata gccattgcgc gtggaccatt 1140 gtgcgcgtgg aaattctgcg caacttttat tttattaacc gcctgaccgg ctatctgcgc 1200 aactaa 1206 <210> 142 <211> 1377 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV-INF-beta1b <400> 142 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgcatcatc atcaccacca cgaaaacctg tatttccaga gctataacct gctgggcttt 900 ctgcagcgca gcagcaactt tcagagccag aaactgctgt ggcagctgaa cggccgcctg 960 gaatattgcc tgaaagatcg catgaacttt gatattccgg aagaaattaa acagctgcag 1020 cagtttcaga aagaagatgc ggcgctgacc atttatgaaa tgctgcagaa catttttgcg 1080 atttttcgcc aggatagcag cagcaccggc tggaacgaaa ccattgtgga aaacctgctg 1140 gcgaacgtgt atcatcagat taaccatctg aaaaccgtgc tggaagaaaa actggaaaaa 1200 gaagatttta cccgcggcaa actgatgagc agcctgcatc tgaaacgcta ttatggccgc 1260 attctgcatt atctgaaagc gaaagaatat agccattgcg cgtggaccat tgtgcgcgtg 1320 gaaattctgc gcaactttta ttttattaac cgcctgaccg gctatctgcg caactaa 1377 <210> 143 <211> 828 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO-H6TEV-INF-beta1b <400> 143 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag agctataacc tgctgggctt tctgcagcgc 360 agcagcaact ttcagagcca gaaactgctg tggcagctga acggccgcct ggaatattgc 420 ctgaaagatc gcatgaactt tgatattccg gaagaaatta aacagctgca gcagtttcag 480 aaagaagatg cggcgctgac catttatgaa atgctgcaga acatttttgc gatttttcgc 540 caggatagca gcagcaccgg ctggaacgaa accattgtgg aaaacctgct ggcgaacgtg 600 tatcatcaga ttaaccatct gaaaaccgtg ctggaagaaa aactggaaaa agaagatttt 660 acccgcggca aactgatgag cagcctgcat ctgaaacgct attatggccg cattctgcat 720 tatctgaaag cgaaagaata tagccattgc gcgtggacca ttgtgcgcgt ggaaattctg 780 cgcaactttt attttattaa ccgcctgacc ggctatctgc gcaactaa 828 <210> 144 <211> 261 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for Insulin glargine precursor (INSG) <400> 144 tttgtgaacc agcatctgtg cggcagccat ctggtggaag cgctgtatct ggtgtgcggc 60 gaacgcggct ttttttatac cccgaaaacc cgccgcgaag cggaagatct gcaggtgggc 120 caggtggaac tgggcggcgg cccgggcgcg ggcagcctgc agccgctggc gctggaaggc 180 agcctgcaga aacgcggcat tgtggaacag tgctgcacca gcatttgcag cctgtatcag 240 ctggaaaact attgcggcta a 261 <210> 145 <211> 300 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV-INSG <400> 145 atgcatcatc atcaccacca cgaaaacctg tatttccagt ttgtgaacca gcatctgtgc 60 ggcagccatc tggtggaagc gctgtatctg gtgtgcggcg aacgcggctt tttttatacc 120 ccgaaaaccc gccgcgaagc ggaagatctg caggtgggcc aggtggaact gggcggcggc 180 ccgggcgcgg gcagcctgca gccgctggcg ctggaaggca gcctgcagaa acgcggcatt 240 gtggaacagt gctgcaccag catttgcagc ctgtatcagc tggaaaacta ttgcggctaa 300 300 <210> 146 <211> 531 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV-INSG <400> 146 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag tttgtgaacc agcatctgtg cggcagccat 300 ctggtggaag cgctgtatct ggtgtgcggc gaacgcggct ttttttatac cccgaaaacc 360 cgccgcgaag cggaagatct gcaggtgggc caggtggaac tgggcggcgg cccgggcgcg 420 ggcagcctgc agccgctggc gctggaaggc agcctgcaga aacgcggcat tgtggaacag 480 tgctgcacca gcatttgcag cctgtatcag ctggaaaact attgcggcta a 531 <210> 147 <211> 684 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV-INSG <400> 147 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagtttgtga accagcatct gtgcggcagc catctggtgg aagcgctgta tctggtgtgc 480 ggcgaacgcg gcttttttta taccccgaaa acccgccgcg aagcggaaga tctgcaggtg 540 ggccaggtgg aactgggcgg cggcccgggc gcgggcagcc tgcagccgct ggcgctggaa 600 ggcagcctgc agaaacgcgg cattgtggaa cagtgctgca ccagcatttg cagcctgtat 660 cagctggaaa actattgcgg ctaa 684 <210> 148 <211> 819 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV-INSG <400> 148 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagtt tgtgaaccag catctgtgcg gcagccatct ggtggaagcg 600 ctgtatctgg tgtgcggcga acgcggcttt ttttataccc cgaaaacccg ccgcgaagcg 660 gaagatctgc aggtgggcca ggtggaactg ggcggcggcc cgggcgcggg cagcctgcag 720 ccgctggcgc tggaaggcag cctgcagaaa cgcggcattg tggaacagtg ctgcaccagc 780 atttgcagcc tgtatcagct ggaaaactat tgcggctaa 819 <210> 149 <211> 885 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV-INSG <400> 149 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagtttgtg aaccagcatc tgtgcggcag ccatctggtg 660 gaagcgctgt atctggtgtg cggcgaacgc ggcttttttt ataccccgaa aacccgccgc 720 gaagcggaag atctgcaggt gggccaggtg gaactgggcg gcggcccggg cgcgggcagc 780 ctgcagccgc tggcgctgga aggcagcctg cagaaacgcg gcattgtgga acagtgctgc 840 accagcattt gcagcctgta tcagctggaa aactattgcg gctaa 885 <210> 150 <211> 969 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV-INSG <400> 150 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagtt tgtgaaccag 720 catctgtgcg gcagccatct ggtggaagcg ctgtatctgg tgtgcggcga acgcggcttt 780 ttttataccc cgaaaacccg ccgcgaagcg gaagatctgc aggtgggcca ggtggaactg 840 ggcggcggcc cgggcgcggg cagcctgcag ccgctggcgc tggaaggcag cctgcagaaa 900 cgcggcattg tggaacagtg ctgcaccagc atttgcagcc tgtatcagct ggaaaactat 960 tgcggctaa 969 <210> 151 <211> 1140 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV-INSG <400> 151 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgcatcatc atcaccacca cgaaaacctg tatttccagt ttgtgaacca gcatctgtgc 900 ggcagccatc tggtggaagc gctgtatctg gtgtgcggcg aacgcggctt tttttatacc 960 ccgaaaaccc gccgcgaagc ggaagatctg caggtgggcc aggtggaact gggcggcggc 1020 ccgggcgcgg gcagcctgca gccgctggcg ctggaaggca gcctgcagaa acgcggcatt 1080 gtggaacagt gctgcaccag catttgcagc ctgtatcagc tggaaaacta ttgcggctaa 1140 1140 <210> 152 <211> 591 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO-H6TEV-INSG <400> 152 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag tttgtgaacc agcatctgtg cggcagccat 360 ctggtggaag cgctgtatct ggtgtgcggc gaacgcggct ttttttatac cccgaaaacc 420 cgccgcgaag cggaagatct gcaggtgggc caggtggaac tgggcggcgg cccgggcgcg 480 ggcagcctgc agccgctggc gctggaaggc agcctgcaga aacgcggcat tgtggaacag 540 tgctgcacca gcatttgcag cctgtatcag ctggaaaact attgcggcta a 591 <210> 153 <211> 573 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for Human growth hormon (hGH) <400> 153 tttccaacca ttccactgtc tcgcctgttt gataatgcaa tgctgcgtgc acatcgtctg 60 catcagctgg catttgatac ctatcaggaa tttgaagaag catatattcc aaaagaacag 120 aaatattctt tcctgcaaaa tccacagacc tctctgtgtt tttctgaatc tattccaacc 180 ccatccaatc gtgaagagac ccaacagaaa tctaatctgg aactgctgcg tatctctctg 240 ttactgattc agtcttggct ggaaccagtg cagttcctgc gttctgtgtt cgcaaattct 300 ctggtttatg gtgcatctga ttctaatgtt tatgatctgc tgaaagattt agaggaagga 360 attcagaccc tgatgggtcg tctggaagat ggttctccac gtaccggtca gatttttaaa 420 cagacctatt ctaaatttga taccaattct cataatgacg atgcactgct gaaaaactat 480 ggtctgctgt actgctttcg taaagatatg gataaagttg aaacctttct gcgtattgtt 540 cagtgtcgtt ctgttgaagg ttcttgtggt ttt 573 <210> 154 <211> 612 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV-hGH <400> 154 atgcatcatc atcaccacca cgaaaacctg tatttccagt ttccaaccat tccactgtct 60 cgcctgtttg ataatgcaat gctgcgtgca catcgtctgc atcagctggc atttgatacc 120 tatcaggaat ttgaagaagc atatattcca aaagaacaga aatattcttt cctgcaaaat 180 ccacagacct ctctgtgttt ttctgaatct attccaaccc catccaatcg tgaagagacc 240 caacagaaat ctaatctgga actgctgcgt atctctctgt tactgattca gtcttggctg 300 gaaccagtgc agttcctgcg ttctgtgttc gcaaattctc tggtttatgg tgcatctgat 360 tctaatgttt atgatctgct gaaagattta gaggaaggaa ttcagaccct gatgggtcgt 420 ctggaagatg gttctccacg taccggtcag atttttaaac agacctattc taaatttgat 480 accaattctc ataatgacga tgcactgctg aaaaactatg gtctgctgta ctgctttcgt 540 aaagatatgg ataaagttga aacctttctg cgtattgttc agtgtcgttc tgttgaaggt 600 tcttgtggtt tt 612 <210> 155 <211> 843 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV-hGH <400> 155 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag tttccaacca ttccactgtc tcgcctgttt 300 gataatgcaa tgctgcgtgc acatcgtctg catcagctgg catttgatac ctatcaggaa 360 tttgaagaag catatattcc aaaagaacag aaatattctt tcctgcaaaa tccacagacc 420 tctctgtgtt tttctgaatc tattccaacc ccatccaatc gtgaagagac ccaacagaaa 480 tctaatctgg aactgctgcg tatctctctg ttactgattc agtcttggct ggaaccagtg 540 cagttcctgc gttctgtgtt cgcaaattct ctggtttatg gtgcatctga ttctaatgtt 600 tatgatctgc tgaaagattt agaggaagga attcagaccc tgatgggtcg tctggaagat 660 ggttctccac gtaccggtca gatttttaaa cagacctatt ctaaatttga taccaattct 720 cataatgacg atgcactgct gaaaaactat ggtctgctgt actgctttcg taaagatatg 780 gataaagttg aaacctttct gcgtattgtt cagtgtcgtt ctgttgaagg ttcttgtggt 840 ttt 843 <210> 156 <211> 996 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV-hGH <400> 156 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagtttccaa ccattccact gtctcgcctg tttgataatg caatgctgcg tgcacatcgt 480 ctgcatcagc tggcatttga tacctatcag gaatttgaag aagcatatat tccaaaagaa 540 cagaaatatt ctttcctgca aaatccacag acctctctgt gtttttctga atctattcca 600 accccatcca atcgtgaaga gacccaacag aaatctaatc tggaactgct gcgtatctct 660 ctgttactga ttcagtcttg gctggaacca gtgcagttcc tgcgttctgt gttcgcaaat 720 tctctggttt atggtgcatc tgattctaat gtttatgatc tgctgaaaga tttagaggaa 780 ggaattcaga ccctgatggg tcgtctggaa gatggttctc cacgtaccgg tcagattttt 840 aaacagacct attctaaatt tgataccaat tctcataatg acgatgcact gctgaaaaac 900 tatggtctgc tgtactgctt tcgtaaagat atggataaag ttgaaacctt tctgcgtatt 960 gttcagtgtc gttctgttga aggttcttgt ggtttt 996 <210> 157 <211> 1131 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV-hGH <400> 157 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagtt tccaaccatt ccactgtctc gcctgtttga taatgcaatg 600 ctgcgtgcac atcgtctgca tcagctggca tttgatacct atcaggaatt tgaagaagca 660 tatattccaa aagaacagaa atattctttc ctgcaaaatc cacagacctc tctgtgtttt 720 tctgaatcta ttccaacccc atccaatcgt gaagagaccc aacagaaatc taatctggaa 780 ctgctgcgta tctctctgtt actgattcag tcttggctgg aaccagtgca gttcctgcgt 840 tctgtgttcg caaattctct ggtttatggt gcatctgatt ctaatgttta tgatctgctg 900 aaagatttag aggaaggaat tcagaccctg atgggtcgtc tggaagatgg ttctccacgt 960 accggtcaga tttttaaaca gacctattct aaatttgata ccaattctca taatgacgat 1020 gcactgctga aaaactatgg tctgctgtac tgctttcgta aagatatgga taaagttgaa 1080 acctttctgc gtattgttca gtgtcgttct gttgaaggtt cttgtggttt t 1131 <210> 158 <211> 1197 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV-hGH <400> 158 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagtttcca accattccac tgtctcgcct gtttgataat 660 gcaatgctgc gtgcacatcg tctgcatcag ctggcatttg atacctatca ggaatttgaa 720 gaagcatata ttccaaaaga acagaaatat tctttcctgc aaaatccaca gacctctctg 780 tgtttttctg aatctattcc aaccccatcc aatcgtgaag agacccaaca gaaatctaat 840 ctggaactgc tgcgtatctc tctgttactg attcagtctt ggctggaacc agtgcagttc 900 ctgcgttctg tgttcgcaaa ttctctggtt tatggtgcat ctgattctaa tgtttatgat 960 ctgctgaaag atttagagga aggaattcag accctgatgg gtcgtctgga agatggttct 1020 ccacgtaccg gtcagatttt taaacagacc tattctaaat ttgataccaa ttctcataat 1080 gacgatgcac tgctgaaaaa ctatggtctg ctgtactgct ttcgtaaaga tatggataaa 1140 gttgaaacct ttctgcgtat tgttcagtgt cgttctgttg aaggttcttg tggtttt 1197 <210> 159 <211> 1281 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV-hGH <400> 159 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagtt tccaaccatt 720 ccactgtctc gcctgtttga taatgcaatg ctgcgtgcac atcgtctgca tcagctggca 780 tttgatacct atcaggaatt tgaagaagca tatattccaa aagaacagaa atattctttc 840 ctgcaaaatc cacagacctc tctgtgtttt tctgaatcta ttccaacccc atccaatcgt 900 gaagagaccc aacagaaatc taatctggaa ctgctgcgta tctctctgtt actgattcag 960 tcttggctgg aaccagtgca gttcctgcgt tctgtgttcg caaattctct ggtttatggt 1020 gcatctgatt ctaatgttta tgatctgctg aaagatttag aggaaggaat tcagaccctg 1080 atgggtcgtc tggaagatgg ttctccacgt accggtcaga tttttaaaca gacctattct 1140 aaatttgata ccaattctca taatgacgat gcactgctga aaaactatgg tctgctgtac 1200 tgctttcgta aagatatgga taaagttgaa acctttctgc gtattgttca gtgtcgttct 1260 gttgaaggtt cttgtggttt t 1281 <210> 160 <211> 1452 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV-hGH <400> 160 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgcatcatc atcaccacca cgaaaacctg tatttccagt ttccaaccat tccactgtct 900 cgcctgtttg ataatgcaat gctgcgtgca catcgtctgc atcagctggc atttgatacc 960 tatcaggaat ttgaagaagc atatattcca aaagaacaga aatattcttt cctgcaaaat 1020 ccacagacct ctctgtgttt ttctgaatct attccaaccc catccaatcg tgaagagacc 1080 caacagaaat ctaatctgga actgctgcgt atctctctgt tactgattca gtcttggctg 1140 gaaccagtgc agttcctgcg ttctgtgttc gcaaattctc tggtttatgg tgcatctgat 1200 tctaatgttt atgatctgct gaaagattta gaggaaggaa ttcagaccct gatgggtcgt 1260 ctggaagatg gttctccacg taccggtcag atttttaaac agacctattc taaatttgat 1320 accaattctc ataatgacga tgcactgctg aaaaactatg gtctgctgta ctgctttcgt 1380 aaagatatgg ataaagttga aacctttctg cgtattgttc agtgtcgttc tgttgaaggt 1440 tcttgtggtt tt 1452 <210> 161 <211> 903 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO-H6TEV-hGH <400> 161 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag tttccaacca ttccactgtc tcgcctgttt 360 gataatgcaa tgctgcgtgc acatcgtctg catcagctgg catttgatac ctatcaggaa 420 tttgaagaag catatattcc aaaagaacag aaatattctt tcctgcaaaa tccacagacc 480 tctctgtgtt tttctgaatc tattccaacc ccatccaatc gtgaagagac ccaacagaaa 540 tctaatctgg aactgctgcg tatctctctg ttactgattc agtcttggct ggaaccagtg 600 cagttcctgc gttctgtgtt cgcaaattct ctggtttatg gtgcatctga ttctaatgtt 660 tatgatctgc tgaaagattt agaggaagga attcagaccc tgatgggtcg tctggaagat 720 ggttctccac gtaccggtca gatttttaaa cagacctatt ctaaatttga taccaattct 780 cataatgacg atgcactgct gaaaaactat ggtctgctgt actgctttcg taaagatatg 840 gataaagttg aaacctttct gcgtattgtt cagtgtcgtt ctgttgaagg ttcttgtggt 900 ttt 903 <210> 162 <211> 111 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for Oxyntomodulin (OXM) <400> 162 catagccagg gcacctttac cagcgattat agcaaatatc tggatagccg ccgcgcgcag 60 gattttgtgc agtggctgat gaacaccaaa cgcaaccgca acaacattgc g 111 <210> 163 <211> 150 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for H6TEV-OXM <400> 163 atgcatcatc atcaccacca cgaaaacctg tatttccagc atagccaggg cacctttacc 60 agcgattata gcaaatatct ggatagccgc cgcgcgcagg attttgtgca gtggctgatg 120 aacaccaaac gcaaccgcaa caacattgcg 150 <210> 164 <211> 381 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMC-H6TEV-OXM <400> 164 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatcatcat 240 catcaccacc acgaaaacct gtatttccag catagccagg gcacctttac cagcgattat 300 agcaaatatc tggatagccg ccgcgcgcag gattttgtgc agtggctgat gaacaccaaa 360 cgcaaccgca acaacattgc g 381 <210> 165 <211> 534 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB1-H6TEV-OXM <400> 165 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacat catcatcacc accacgaaaa cctgtatttc 420 cagcatagcc agggcacctt taccagcgat tatagcaaat atctggatag ccgccgcgcg 480 caggattttg tgcagtggct gatgaacacc aaacgcaacc gcaacaacat tgcg 534 <210> 166 <211> 669 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCB2-H6TEV-OXM <400> 166 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttcatcatca tcaccaccac 540 gaaaacctgt atttccagca tagccagggc acctttacca gcgattatag caaatatctg 600 gatagccgcc gcgcgcagga ttttgtgcag tggctgatga acaccaaacg caaccgcaac 660 aacattgcg 669 <210> 167 <211> 735 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC1-H6TEV-OXM <400> 167 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatca tcatcatcac 600 caccacgaaa acctgtattt ccagcatagc cagggcacct ttaccagcga ttatagcaaa 660 tatctggata gccgccgcgc gcaggatttt gtgcagtggc tgatgaacac caaacgcaac 720 cgcaacaaca ttgcg 735 <210> 168 <211> 819 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCC2-H6TEV-OXM <400> 168 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtcatcatca tcaccaccac gaaaacctgt atttccagca tagccagggc 720 acctttacca gcgattatag caaatatctg gatagccgcc gcgcgcagga ttttgtgcag 780 tggctgatga acaccaaacg caaccgcaac aacattgcg 819 <210> 169 <211> 990 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for IMCD1-H6TEV-OXM <400> 169 atggccggaa aaagcagtac tgaaaagaaa tatattgtcg gatttaagca gactatgagt 60 gccatgagtt ccgccaagaa aaaggatgtt atttctgaaa aaggcggaaa ggttcaaaag 120 caatttaagt atgttaacgc ggccgcagca actttggatg aaaaagctgt aaaagaattg 180 aaaaaagatc cgagcgttgc ttatgtggaa gaagatcata ttgctcatga atatgcgcaa 240 agctttcctt atggcatttc tcaaattaaa gcgccggctc ttcactctca aggctatact 300 ggctctaacg taaaagtagc tgttatcgat agcggaattg actcttctca tcctgactta 360 aacgtccgtg gcggagcaag cttcgtacct tctgaaacta acccatacca ggacggcagt 420 tctcacggta cgcatgtagc cggtacgatt gccgctctta ataactcaat cggtgttctg 480 ggcgtagcgc caagcgcatc attatatgca gtaaaagtgc ttgattcaac tggaagcggc 540 caatatagct ggattattaa cggcattgag tgggccattt ccaataatat ggatgttatc 600 aatatgagcc ttggcggacc tactggttct actgcgctga aaactgtcgt tgataaagcc 660 gtttccagcg gtatcgtcgt tgctgccgca gccggaaacg aaggttcatc cggaagcact 720 agcactgtcg gctaccctgc aaaatatcct tctactattg cagtaggtgc ggtaaatagc 780 agcaaccaac gtgcttcatt ctccagcgca ggttctgagc ttgatgtgat ggctcctggc 840 gtgcatcatc atcaccacca cgaaaacctg tatttccagc atagccaggg cacctttacc 900 agcgattata gcaaatatct ggatagccgc cgcgcgcagg attttgtgca gtggctgatg 960 aacaccaaac gcaaccgcaa caacattgcg 990 <210> 170 <211> 441 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for SUMO-H6TEV-OXM <400> 170 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct 60 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa 120 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg 180 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat 240 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg tggtcatcat 300 catcaccacc acgaaaacct gtatttccag catagccagg gcacctttac cagcgattat 360 agcaaatatc tggatagccg ccgcgcgcag gattttgtgc agtggctgat gaacaccaaa 420 cgcaaccgca acaacattgc g 441 <210> 171 <211> 4271 <212> DNA <213> Artificial Sequence <220> <223> nucleotide sequence for expression vector(plasmid) <400> 171 actgcataat tcgtgtcgct caaggcgcac tcccgttctg gataatgctt tttgcgccga 60 catcataacg gttctggcaa atattctgaa atgagctgtt gacaattaat catccggctc 120 gtataatgtg gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac 180 tttaagaagg agatatacat atgaaatacc tgctgccgac cgctgctgct ggtctgctgc 240 tcctcgctgc ccagccggcg atggccatgg atatcggaat taattcggat ccgaattcga 300 gctccgtcga caagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg 360 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag 420 cataacccct tggggcctct aaacgggtct tgaggggttt tttgagcggt ctgataaaac 480 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag 540 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc 600 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt 660 ttgtcggtga acgctctccc gagtaggaca aatccgccgg gagcggattt gaacgttgcg 720 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt 780 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt 840 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 900 ataattccaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg ggttgcacgc 960 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 1020 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 1080 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 1140 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 1200 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatccc accttgctcc 1260 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 1320 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 1380 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 1440 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 1500 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 1560 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 1620 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 1680 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgaa ttgaaaaagg 1740 aagagtatga gtattcaaca tttccaagaa aaggatctag gtgaagatcc tttttgataa 1800 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 1860 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 1920 aaaaaaacca ccgctaccaa cggtggtttg tttgccggat caagagctac caactctttt 1980 tccgaaggta actggcttca gcagagcaca gataccaaat actgttcttc tagtgtagcc 2040 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 2100 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 2160 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 2220 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 2280 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca ggatcggaac 2340 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 2400 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 2460 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 2520 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 2580 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 2640 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg 2700 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc 2760 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg 2820 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg 2880 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca 2940 attcgcgcgc gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac 3000 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa 3060 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg 3120 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat 3180 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt 3240 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc 3300 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag 3360 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct 3420 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa 3480 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc 3540 ccatgaaaac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat 3600 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca 3660 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc 3720 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat 3780 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct 3840 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta 3900 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga 3960 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc 4020 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt 4080 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc 4140 gcaacgcaat taatgtaagt tagcgcgaat tgatctggtt tgacagctta tcatcgactg 4200 cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca 4260 ggtcgtaaat c 4271

Claims (13)

  1. 서열번호 1, 서열번호 2, 서열번호 3, 서열번호 4, 서열번호 5 및 서열번호 6으로 이루어진 군에서 선택되는 어느 하나의 아미노산 서열로 이루어진 융합 태그; 링커; 서열번호 9의 아미노산 서열로 이루어진 담배 식각 바이러스 프로테아제 인식 서열; 옥신토모듈린(oxyntomodulin, OXM);가 N-말단에서 C-말단의 방향으로 순서대로 위치하는 융합폴리펩타이드를 코딩하는 폴리뉴클레오타이드를 포함하는 발현벡터를 제조하는 단계 (a);
    상기 단계 (a) 후, 상기 발현벡터를 대장균에 도입하여 형질전환 대장균을 제조하는 단계 (b);
    상기 단계 (b) 후, 상기 형질전환 대장균을 배양하는 단계 (c);
    상기 단계 (c) 후, 상기 형질전환 대장균에서 봉입체 형태로 발현된 융합 폴리펩타이드를 분리하고 가용화하는 단계 (d); 및
    상기 단계 (d) 후, 상기 가용화된 융합 폴리펩타이드에 담배 식각 바이러스 프로테아제를 처리하여 옥신토모듈린(oxyntomodulin, OXM)을 회수하는 단계 (e);를 포함하는 것을 특징으로 하는 옥신토모듈린(oxyntomodulin, OXM)의 생산방법.
  2. 삭제
  3. 삭제
  4. 삭제
  5. 제1항에 있어서,
    상기 링커는,
    히스티딘 친화성 태그를 포함하는 것을 특징으로 하는, 옥신토모듈린(oxyntomodulin, OXM)의 생산방법.
  6. 삭제
  7. 삭제
  8. 삭제
  9. 삭제
  10. 삭제
  11. 삭제
  12. 삭제
  13. 삭제
KR1020200168669A 2019-12-06 2020-12-04 옥신토모듈린 생산용 융합태그 KR102301138B1 (ko)

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