[Expression of isoleucine zipper modified soluble CD40L in Pichia pastoris].
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.
Du
Xiaobo
X
Key Laboratory of Biotherapy of Human Diseases, Ministry of Education, Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
Tian
Ling
L
Wang
Yongsheng
Y
Diao
Peng
P
Wang
Guoqing
G
Wei
Yuquan
Y
D000804:Angiotensin II; D023201:CD40 Ligand / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D007532:Isoleucine / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000235:genetics*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D011995:Recombination, Genetic
D011993:Recombinant Fusion Proteins; D007532:Isoleucine; D000804:Angiotensin II; D023201:CD40 Ligand