Basic study of gene therapy for severe pain by using virus vector

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 17591612
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Anesthesiology/Resuscitation studies
• Research Institution: University of Tsukuba
• Principal Investigator: INOMATA Shinichi University of Tsukuba, Graduate School of Comprehensive Human Sciences, Associate Professor (10282352)
• Co-Investigator(Kenkyū-buntansha): MIYOSHI Hiroyuki RIKEN Bio Resource Center, Subteam for Manipulation of Cell Fate, Team leader (70219830)
• Project Period (FY): 2005 – 2007
• Keywords: severe pain / enkephalin / virus vector / gene therapy

Research Abstract

Total RNA of a human adrenal gland was obtained from patients, received surgical operation, with their informed consents. The cDNA was synthesized by reverse transcriptase using a primer. The pre-pro-enkephalin (PPE) gene was then amplified by polymerase chain reaction with specific upstream and downstream oligoneucleotides. The polymerase chain reaction product contained the full-length PPE coding cDNA. The amplified fragment was cloned into a vector (pDON-AI-2-Neo or pMEI-5 Neo [Not I-BamHI]). The plasmid DNA was purified and digested with restriction enzymes and electrophoresed on agarose gel. The higher efficacy of infection was observed in pDON-AI-2-Neo.