[The Prediction of the Molecular Biology Characteristics of Recombinant Fusion Exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40]
In order to confirm the reasonability of designed recombinant exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40, their molecular biology characteristics, such as flexibility, antigenicity, hydrophilicity, epitope and secondary structure, were predicted by using a computer software GOLDKEY. It had been found that the recombinant fusion exotoxin had kept the epitope characterstics of B7-1, B7-2 and PE40, and had not got new epitope, and the antigenicity in flexible linker was extxemely low. The linker inserted in the recombinant fusion exotoxin had low epitope, low antigenicity and high flexibility. Compared to B7-1, B7-2 and PE40, there are several amino acid residues changes in B7-1-Linker-PE40 and B7-2-Linker-PE40, respectively, which might have some effect on secondary structure of the recombinant fusion exotoxins. Western blot analysis revealed that both B7-1-Linker-PE40 and B7-2-Linker-PE40 could bind specifically with antibodies against B7-1, B7-2 and PE40, respectively. The result of Western blot was consistant with the computer prediction that the recombinant proteins retain the antigenicity and spacial structure of B7 and PE40. It is suggested that both fusion proteins designed and constructed were resonable and computer analysis would be helpful for us to study the biological activity of the recombinant fusion exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40 and construct other recombinant proteins further.
Zhang
Hui-Li
HL
Department of Immunology, The Affilicated Hospital, Academy of Military Medical Sciences, Beijing 1000391, China; Shenzhen Baoan Central Station of Blood Transfusion, Shenzhen 5181012, China.
Xi
Yong-Zhi
YZ
Kong
Fan-Hua
FH
Chen
Ru-Guang
RG
Yuan
Zhi-Hong
ZH
Liu
Nan
N
Guan
Hai-Rong
HR