Sub-cloning and soluble indution of CILP-MBP recombinant protein by tempera-ture variable / 中国免疫学杂志
Chinese Journal of Immunology
;
(12)1985.
Artigo
em Chinês
| WPRIM
| ID: wpr-536893
ABSTRACT
Abstract Objective:
To construct the Second half of CILP(C2) MBP fusion protein by sub-cloning technology.Methods:
Recombinantfusion proteins, which contain the fragments within the C2 region(designated C2F1, C2F2 and C2F3) of the non-porcine nucleotide pyrophos-phohydrolase-homologous region of CILP, were prepared using pMAL-eHis vector. The recombinat genes are induced by different temperatures(22℃,30℃,37℃ ).Results:
Expression using pMAL-eHis system can be induced chemically by adding IPTG. 37℃ temperature prmotes in-soluble inclusion-body formation,but 22℃ temperature can not induce the enough expression of recombinant protein. Onl 30℃ temperaturecan induce enough amount of soluble recombinant protein. The characers of fusion proteins that they carried 6 straight histidines, (His)6, at tbeC-terminus of multiple cloning sites for affinity purification were assessed by sodium dodecy1 sulfate-polyacylamide gel electrophoresis(SDS-PAGE) and Western blot. Nucleotide sequences of the insertion genes were confirmed by dideoxy sequencing.Conclusion:
C2F1, C2F2,C2F3-MBP fnsion proteins were constructed successfully.These recombinant proteins may provide important roles in the future study on CILP.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Chinese Journal of Immunology
Ano de publicação:
1985
Tipo de documento:
Artigo
Similares
MEDLINE
...
LILACS
LIS