Sub-cloning and soluble indution of CILP-MBP recombinant protein by tempera-ture variable
- VernacularTitle:CILP-MBP基因重组蛋白的亚克隆构建及温度可溶性诱生表达
- Author:
Zhenyu YAO
;
Changlong LU
;
Bingzhi YU
- Publication Type:Journal Article
- Keywords:
CILP Recombinmant protein Sub-cloning
- From:
Chinese Journal of Immunology
1985;0(05):-
- CountryChina
- Language:Chinese
-
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.
