title: [Cloning and expression of fusion gene of amyloid beta binding alcohol dehydrogenase decoy peptide aptamer (TRX1-ABAD-DP-TRX2)].
abstract: OBJECTIVE
  To construct TRX-ABAD-DP-TRX (T-A-T) fusion gene of a novel ABAD-DP aptamer through the insertion of ABAD-DP into the modified human thioredoxin (hTRX) and exploit the possibility of further applications for the gene therapy of Alzheimer's disease.
  
  
  METHODS
  According to the designed sequence, the target fragments of TRX1, TRX2 and ABAD-DP were created by PCR (polymerase chain reaction) and then inserted into the multiple clone site of adeno-associated virus shuttle plasmid pSSHG-CMV with gene cloning technique. The corresponding fusion gene TRX1-ABAD-DP-TRX2 was identified by restriction enzymes digestion with EcoRI and BamHI. The recombinant adeno-associated virus (AAV/T-A-T) was produced in HeLa cells with linear polyethylenimine. The expression of T-A-T fusion gene and co-localization between T-A-T and Aβ peptide in NIH 3T3 cells were examined by fluorescent immunohistochemistry.
  
  
  RESULTS
  The size of fusogenic fragment TRX1-ABAD-DP-TRX2 was approximately 435 bp. And it was consistent with our design. T-A-T fusion gene was expressed in NIH 3T3 cells. Through co-expression, T-A-T aptamer and intracellular Aβ peptide were co-localized. It indicated that T-A-T aptamer could bind Aβ within NIH 3T3 cells.
  
  
  CONCLUSIONS
  The TRX1-ABAD-DP-TRX2 fusion gene is successfully cloned and expressed. And it may provide rationales for further applications in the gene therapy of Alzheimer's disease's.
authors.affiliation: Department of Neurology, Jilin University, Changchun, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Wang
authors.forename: Jiang
authors.initials: J
authors.lastname: Wu
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.forename: Ming-yu
authors.initials: MY
authors.lastname: Wang
authors.forename: Li-he
authors.initials: LH
authors.lastname: Che
authors.forename: Yu
authors.initials: Y
authors.lastname: Yang
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics*; D016229:Amyloid beta-Peptides / Q000235:genetics*; D000818:Animals; D052158:Aptamers, Peptide / Q000235:genetics*; D019677:Artificial Gene Fusion; D000229:Dependovirus / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D013879:Thioredoxins / Q000235:genetics
chemical_list: D016229:Amyloid beta-Peptides; D052158:Aptamers, Peptide; D013879:Thioredoxins; D000426:Alcohol Dehydrogenase