Expression of isoleucine zipper modified soluble CD40L in Pichia pastoris.
- Author:
Xiaobo DU
1
;
Ling TIAN
;
Yongsheng WANG
;
Peng DIAO
;
Guoqing WANG
;
Yuquan WEI
Author Information
1. Key Laboratory of Biotherapy of Human Diseases, Ministry of Education, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Angiotensin II;
CD40 Ligand;
biosynthesis;
genetics;
Cloning, Molecular;
Isoleucine;
analogs & derivatives;
biosynthesis;
genetics;
Pichia;
genetics;
metabolism;
Plasmids;
Polymerase Chain Reaction;
methods;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
Recombination, Genetic
- From:
Journal of Biomedical Engineering
2006;23(4):844-847
- CountryChina
- Language:Chinese
-
Abstract:
To obtain the expression of Isoleucine Zipper modified soluble CD40L (IZ-sCD40L) in Pichia pastoris, firstly, DNA fragment of IZ-sCD40L was obtained by PCR and over-lap PCR . Then the expression vector pPICZaA-IZ-sCD40L was constructed. Nucleotide sequencing analysis indicated that the DNA fragment of IZ-sCD40L was correctly inserted into the pPICZaA vector. Linearized pPICZ(alpha)A-IZ-sCD40L was introduced into Pichia pastoris GS115. Positive clone was selected by PCR and its phenotype was determined. The positive clone was introduced with methanol. The results of SDS-PAGE and Western blot showed that product was recombinant Isoleucine Zipper modified soluble CD40L fusion protein.
