Journal: Translational Cancer Research
Article Title: Fusion of NY-ESO-1 epitope with heat shock protein 70 enhances its induced immune responses and antitumor activity against glioma in vitro
doi: 10.21037/tcr-23-1476
Figure Lengend Snippet: Construction of pET30a -HSP70 vectors and recombinant protein expression and purification. (A) Schematic representation of the recombinant pET30a- HSP70 vectors. The full-length HSP70 gene was cloned into the vector pET30a using the restriction sites EcoR1 and Hind III. (B) RT-PCR confirmation of the recombinant HSP70 . Lane 1: the full-length HSP70 . Lane 2: double enzyme-digested 2,000-bp gene fragment. (C) HSP70 protein expressed by recombinant vector pET30 -HSP70 . Lane M: protein molecular weight markers. Lanes 1–2: whole cell lysate of E. coli Rosetta 2 transformed with pET30- HSP70 without [1] and with [2] induction of IPTG. Lane 3: purified HSP70 protein at 1× loading concentration. HSP70, heat shock protein 70; RT-PCR, reverse transcription-polymerase chain reaction; IPTG, isopropyl β-d-thiogalactoside.
Article Snippet: Prokaryotic expression plasmid pET30a (Promega, Madison, WI, USA) was kept in our laboratory.
Techniques: Recombinant, Expressing, Purification, Clone Assay, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Molecular Weight, Transformation Assay, Concentration Assay, Reverse Transcription, Polymerase Chain Reaction