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Analysis of interaction among proteins expressed in retinal pigment epithelium

Research Project

Project/Area Number 17591818
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Ophthalmology
Research InstitutionNippon 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)
原 宏二  秋田大学, 医学部, 助手 (60375251)
Project Period (FY) 2005 – 2006
Project Status Completed (Fiscal Year 2006)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2005: ¥2,400,000 (Direct Cost: ¥2,400,000)
KeywordsMfrp / Clqtnf5 / C1qtnf5 / rd6マウス
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.

Report

(3 results)
  • 2006 Annual Research Report   Final Research Report Summary
  • 2005 Annual Research Report

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Published: 2005-04-01   Modified: 2016-04-21  

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