[Construction and expression of a vector containing protein transduction domain and bcr/abl fusion gene]

Zhonghua Xue Ye Xue Za Zhi. 2002 Jan;23(1):5-8.
[Article in Chinese]

Abstract

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.

MeSH terms

  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Fusion Proteins, bcr-abl / genetics*
  • Fusion Proteins, bcr-abl / metabolism
  • Gene Expression
  • Gene Products, tat / genetics*
  • Gene Products, tat / metabolism
  • Genetic Vectors / genetics
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / genetics
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Gene Products, tat
  • Recombinant Fusion Proteins
  • Fusion Proteins, bcr-abl