1993 Fiscal Year Final Research Report Summary
Cloning and expression of Na^+/I^--symporter
| Project/Area Number |
04671493
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
内分泌・代謝学
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| Research Institution | Miyazaki Medical College |
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Principal Investigator |
KOTANI Tomio Miyazaki Medical College, Laboratory Medicine, Assistant Professor, 医学部, 助教授 (10161936)
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| Project Period (FY) |
1992 – 1993
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| Keywords | Na^+ / I^--symporter / cloning |
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| Research Abstract |
In order to get a cDNA for Na^+/I^--symporter, I constructed a cDNA library using a expression vector, pCDM8, and mRNA with more than 1 kb molecular size from FRTL-5. The library was divided into 120 pools consisting of 500 to 2,000 E.coli colonies. The plasmid purified from each pool was transfected Cos7 by the dextran method to measure ^<125>I-uptake. However, I could not get any cDNAs for Na^+/I^--symporter in this screening system.
Cosequently, I concluded that Na^+/I^--symporter would not express successfully in Cos7.
As a next attempt, I have made a plan that cDNA for Na^+/I^--symporter should be screened by the procedure of Vilijn, i.e.mRNA transcribed from each plasmid pool in vitro should be injected into Xenopus oocytes to measure their ^<125>I uptake. Since I have no tools for microinjection and have had no experience in microinjection. I have been purchasing microinjection tools and practicing microinjection. Therefore, I have no data about Na^+/I^--symporter cDNA.
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