Rapid isolation of tissue-specific genes from rat kidney.
A systematic effort to isolate kidney-specific genes was performed using recently described PCR-select methodology. Using this technique, a kidney-specific mini-gene library was generated and a number of kidney-specific genes that share significant homology to previously characterized kidney genes from rats and other species were isolated. These included three renal-specific transporters (an ADH water channel, the anion transporters RST and ROAT1), a cell adhesion molecule (K-cadherin) and a kidney-specific protein upregulated in renal carcinoma (DD96). In addition, we isolated two novel genes from a rat kidney. One of the genes shares limited homology to rat profilin-1 while the other did not share any similarity to genes in the Genbank. Northern blot analysis revealed that the mRNA for each of these genes is expressed in a highly kidney-restricted fashion. Our results suggested that tissue-specific genes can be rapidly isolated and characterized using PCR-select techniques and this methodology may be generally applicable to isolate specific genes from a variety of tissues.
Hu
E
E
Department of Renal Pharmacology, SmithKline Beecham Pharmaceuticals, King of Prussia, PA 19406, USA. Erding_Hu-1@sbphrd.com
Chen
Z
Z
Fredrickson
T A
TA
Spurr
N
N
Gentle
S
S
Sims
M
M
Zhu
Y
Y
Halsey
W
W
Van Horn S
Mao
J
J
Sathe
G M
GM
Brooks
D P
DP
D000818:Animals; D001483:Base Sequence / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D007668:Kidney / Q000502:physiology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D018076:DNA, Complementary