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Expression pattern in a modified equalized kidney cDNA library of hypertensive rat
T H Chern1, F T Chiang, K D Wu
1Institute of Clinical Medicine, National Yang-Ming University, School of Medicine, Taipei, Taiwan.
Background:
Genes with important functions and rarely expressed would probably more easily be cloned from a modified equalized kidney cDNA library for further investigation.
Methods:
A kidney cDNA library of a spontaneously hypertensive rat was synthesized by a modified equalization method. Inserts of random clones were amplified by PCR and sequenced. Sequences were compared against a nonredundant database in GenBank. The cDNA profile was compared with an expression profile of a mouse renal proximal tubule cDNA library. Seven clones were analyzed by Northern blot analysis. The cDNA ends of two novel genes were amplified by PCR, sequenced and analyzed.
Results:
336 cDNA clones were analyzed and grouped into 323 species of transcript with 77 species similar to previously reported genes. Northern blot analysis identified one kidney-specific, one rarely expressed and lung-specific, and another relatively testis-specific gene. Two novel genes were cloned. One was 4.1 kb in length and encoded a 390-amino acid zinc-finger protein. Another was 2.5 kb and encoded a 474-amino acid protein of unknown function. Compared with the expression profile of a mouse renal proximal tubule cDNA library, this kidney library had a lower proportion of ribosomal genes and had a greater proportion of genes for signal transduction and DNA or RNA binding.
Conclusions:
Rare or novel genes could be more easily isolated from this library for molecular study of hypertension and renal pathophysiology.