High level expression of human Interleukin 6 in E. coli
- VernacularTitle:人白细胞介素6在大肠杆菌中的高效表达
- Author:
Jide TIAN
- Publication Type:Journal Article
- Keywords:
Human IL-6;
Gene expression;
E. coll;
Expression condition
- From:
Chinese Journal of Immunology
1985;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Utilizing Polmerase Chain Reaction (PCR) and recombination DNA techniques with synthetic oligonucleotide primers,a high level expressing clone for human IL- 6, pBMhIL6, was constructed successfully in our laboratory. The intact human rIL-6 protein molecular was expressed under the control of p_Rp_L promoters, synthetic SD sequence and the terminal codon TAA which replaced TAG of original human IL-6 in E. coli. the expressed human rIL-6 protein, molecular wight 21 KDa, with specific binding to Anti-IL-6 McAb, constituted about 28% of the total cellular proteins assessed by SDS-PAGE, densitometry analysis and Western Blot assay. The results of study on kinetics of inducing human rIL-6 expression in the defferent E. coli strains and influence of bacteria growth states ingdicate that the higher productive ratio for human rIL-6 inducing expression was obtainde in E. coli DH5a at O. D=0. 7. 5?10~6 unites of human rIL-6 HPGF bioactivities were produced from 1 litre of bacteria extracts assayed by 3HTdR uptake of 7TD1 cell line.
