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Construction of CTLA-4-Ig Fusion Gene in pBudCE4.1 Expression Vector
Mahsa Yazdanpanah-Samani1, Elham Mahmoudi Maymand1, Tayebeh Jahangeerfam2
1Shiraz Institute for Cancer Research, Faculty of Medicine, Shiraz University of Medical Science, Shiraz, Iran.
Background:
CTLA-4 inhibitory signals prevent cell cycle progression and IL-2 production, leading to a halt on an ongoing immune response. CTLA4-Ig fusion proteins contain the extracellular domain of CTLA-4 and Fc fragment of human IgG antibody. In this study we aimed to fuse the ctla-4 gene encoding the extracellular domain of CTLA-4 molecule with igg1 gene encoding Fc region of human IgG.
Methods:
After primer design, PCR reaction was performed using pfu polymerase enzyme and specific primers. PCR amplified fragment was ligated into the vector containing the human igg1 gene. The resulting fusion fragment of ctla-4 and human igg1 genes was ligated to pBudCE4.1 expression vector.
Results:
Extracellular domain of ctla-4 gene was ligated to igg1 gene and then ctla4-ig fragment was cloned into pBudCE4.1 vector. Construction of the expression vector was confirmed by restriction pattern analysis and sequencing.
Conclusion:
By confirming the construct, in the next step, the recombinant DNA will be used to produce CTLA4-Ig recombinant protein for the clinical uses.
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