Enterokinase is a heterodimeric serine protease consisting of 82~140kDa heavy chain and 35-62kDa light chain. The light chain is a catalytic subunit of enterokinase, which consists of a chymotrypsin-like serine protease. Because of high specificity for the amino-acid sequence (Asp4)Lys, enterokinase has a great potential as a fusion protein cleavage reagent. Recombinant HEKL was produced in transgenic rice cell suspension culture with the rice amylase 3D promoter. RAmy3D promoter is highly expressed by sugar starvation. The expression of recombinant HEKL as mRNA and protein in transgenic rice cell was detected by Northern and Western blot analyses, recombinant HEKL was observed about 43 kDa in Western blot analysis. The activity of recombinant HEKL was determined by cleavage of the fusion protein EGFP-hTNFa as substrate, which contained a enterokinase cleavage site (DDDDK) between two moieties. Under optimal conditions the hTNFa product with expected size of 17kDa was well detected on Western blot analysis. These results demonstrate the use of HEKL derived from plant cell suspension culture as proteases (This Study was supported by a grant from the Ministry of Knowledge Economy).
키워드
EnterokinaseRice amylase 3D promoterPlant cell suspension culture
저자
Eun-Sun JEUNG [ Jeonju Biomaterials Institute, Jeonju 561-360, KOREA. ]
Yun-Ji SHIN [ Jeonju Biomaterials Institute, Jeonju 561-360, KOREA. ]
Eun-Jin CHOI [ Jeonju Biomaterials Institute, Jeonju 561-360, KOREA. ]
Ji-Ae YANG [ Jeonju Biomaterials Institute, Jeonju 561-360, KOREA. ]
Kang-Yeol YU [ Jeonju Biomaterials Institute, Jeonju 561-360, KOREA. ]
Hong-Soo DOO [ Jeonju Biomaterials Institute, Jeonju 561-360, KOREA. ]
Moon-Sik YANG [ Division of Biological Sciences, Chonbuk National University, Jeonju 561-756, KOREA. ]
Tea-Ho KWON [ Jeonju Biomaterials Institute, Jeonju 561-360, KOREA. ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
1. 생물공학 분야의 발전을 위한 연구 협력
2. 생물공학의 실용화를 촉진시키기 위한 산학 협동
3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최
4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간
5. 생물공학 발전을 위한 정책 건의
6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동