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[Construction and expression of a vector containing protein transduction domain and bcr/abl fusion gene]
Yingmin Liang1, Qiang Sun, Shanshan Jiang
1Department of Hematology of Tangdu Hospital and Department of Biochemistry and Molecular Biology, The Fourth Military Medical University, Xi'an 710038, China.
Objective:
To construct a vector containing protein transduction domain (PTD) and bcr/abl fusion gene of chronic myelogenous leukemia and express PTD-bcr/abl fusion protein in E. Coli.
Methods:
DNA fragment encoding PTD was synthesized and fused to PCR-amplified bcr/abl gene fragment, then inserted into plasmid pET-16b to get the expression vector pEPb containing PTD-bcr/abl fusion gene, which was transfected and expressed in E. Coli LB21. PTD-bcr/abl fusion protein was purified by affinity chromatography.
Results:
523 bp bcr/abl fusion gene was effectively amplified. The PTD-bcr/abl gene sequencing showed the same sequence as scheduled. The fusion peptide was successfully expressed in E. Coli and purified.
Conclusion:
The results may provide a new PTD-bcr/abl fusion peptide for the immunotherapy of CML.