|Budget Amount *help
¥2,000,000 (Direct Cost : ¥2,000,000)
Fiscal Year 1993 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 1992 : ¥1,300,000 (Direct Cost : ¥1,300,000)
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.