Analysis of interaction among proteins expressed in retinal pigment epithelium

2006 Fiscal Year Final Research Report Summary

• Project/Area Number: 17591818
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Ophthalmology
• Research Institution: Nippon Medical School
• Principal Investigator: KAMEYA Shuhei Nippon Medical School, Faculty of Medicine, Research Associate, 医学部, 助手 (30302269)
• Co-Investigator(Kenkyū-buntansha): YAMAKI Kunihiko Nippon Medical School, Faculty of Medicine, Associate Professor, 医学部, 助教授 (20125751)
• Project Period (FY): 2005 – 2006
• Keywords: Mfrp / Clqtnf5

Research Abstract

To elucidate the interaction between MFRP and CTRP5, we have made antibodies against these proteins. We have already made two different antibodies against MFRP. We made another antibody that recognize different antigenic site of MFRP and antibody against CTRP5 this year, to analyze the interaction more precisely. To express MFRP and CTRP5 in vivo, we have cloned cDNA of Mfrp gene and Clqtnf5 gene of mice respectively. We amplified those cDNA by using high-fidelity DNA polymerase. cDNA sequence has confirmed by sequencing and there was no mutation. We have also cloned cDNA of Mfrp with truncated mutation that is responsible for the phenotype of rd6 mice. We have subcloned the cDNA to pET-17b vector. We have been trying to express those proteins by using PURE SYSTEM classic mini that is in vitro protein-expression system. However, this system does not work well so far. We keep trying several trouble shooting methods.