Expression cloning from plasmid library

Michael Romero MROMERO at
Wed Apr 19 14:31:48 EST 1995

Hi Netters,
	there was a query about expression cloning from a plasmid
library.  This has been used successfully by several groups...
in fact, EC using Xenopus laevis oocytes (after in vitro transcription
from linearized plasmids) was pioneered by Matthias A. Hediger, who
is now at Brigham & Women's Hospital / Harvard in Boston.

1. first paper
	Hediger et al. Nature 330:379-381, 1987
	this is the expression cloning of the Na+/glucose co-transporter
from a rabbit small intestine plasmid cDNA library.

2.  Kanai and Hediger. Nature 360:367-471, 1992 ==>electrogenic high-affinity,
	glutamate transporter, EAAC1

3. Wells and Hediger. PNAS, USA 89: 5596-5600, 1992==> neutral and dibasic
amino acid transporter, D2

4.  Fei et al. Nature 368: 563-566, 1994 ==> proton-coupled oligopeptide

5.  Ho et al. Nature   362:31-38, 1993 ==> inward rectifying ATP-regulated
   K+ channel

6.  Gamba, et al.  PNAS, USA 90:2749-2753, 1993 ==> thiazide-sensitive
    NaCl co-transporter

7.  Brown, et al  Nature 366: 575-580, 1993 ==> Ca2+ sensing receptor

just to list a few that i'm immediately familiar with

8.  also Na\PO4 cotansporter and Na/SO4 cotransporter(s) from Heini Murer's
lab in Switzerland...i don't have those refs at my finger tips.

anyway, lots of people have done it / are continuing to do it.  a variety of
plasmid vectors have been used... pBluescript, pSport, just off the top of
my head.  try doing a medline search  on "expression cloning" to get a
more complete list.  Methods are probably outlined in some protocol book
by now.

good luck,
mromero at

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