The expression and antigenicity identification of recombinant rat TGF-β1 in bacteria  

作　　者：GaoCF KongXT 

机构地区：[1]DepartmentofLaboratoryMedicine,ChangzhengHospital,SecondMilitaryMedicalUniversity,Shanghai200003,China [2]DepartmentofLaboratoryMedicine,ChangzhengHospital,SecondMilitaryMedicalUniversity,Shanghai200003,

出　　处：《Cell Research》2001年第2期95-100,共6页细胞研究（英文版）

基　　金：Shanghai Medical Development grant No. ZD99001 and aGrant (SFB-542) from the Deutsche Forschungsgemeinschaft.

摘　　要：In order to study structure-function details of TGF-beta1, the recombinant mature form of rat TGF-beta1 was expressed in bacteria. Synthesis of the 112 amino-acid carboxyl-terminal part of TGF-beta1 (amino acid 279-390) was controlled by an inducible gene expression system based on bacteriophage T7 RNA polymerase. This system allowed an active and selective synthesis of recombinant TGF-beta1. The molecular weight of expressed TGF-alpha1 monomer determined on SDS-polyacrylamide gel under reducing conditions was about 13 kD. Serial detergent washes combined with a single gel-filtration purification step were sufficient to purify the expression product to homogeneity. Amino-terminal sequencing revealed that the N-terminal of the recombinant protein was identical to the published data. In Western blot analysis the recombinant polypeptide showed excellent antigenicity against polyclonal TGF-beta1 antibody. The mature recombinant rat TGF-beta1 expressed in this study provides a useful tool for future detailed structural and functional studies.

关 键 词：Amino Acid Sequence Animals Base Sequence EPITOPES Escherichia coli Gene Expression Regulation  Bacterial Genetic Vectors Molecular Sequence Data Plasmids Protein Structure  Tertiary Rats Recombinant Proteins Research Support  Non-U.S. Gov't Transformation  Genetic Transforming Growth Factor beta 

分 类 号：Q516[生物学—生物化学] R392.11[医药卫生—免疫学]