[Novel full-length cDNA cloning from normal adrenal gland and pheochromocytoma and functional prediction].
OBJECTIVE
To investigate methodology of cloning full-length cDNA from tissues of normal adrenal gland and pheochromocytoma and predict their function.


METHODS
104 samples from normal adrenal gland and 22 samples from pheochromocytoma were examined by expressed sequence tags (EST) sequencing, bioinformatics analysis, in silico cloning, rapid amplification of cDNA ends and RT-PCR.


RESULTS
Among the 126 samples of novel full-length cDNA cloning 104 were from normal adrenal gland, and 22 from pheochromocytoma (PC). Among the novel genes 50 were cloned by direct sequencing of clones, 74 by in silico cloning, and 2 by rapid amplification of cDNA ends, and the localization of chromosomes was made with UniGene searching and radiation hybrid (RH) procedure. Comparison of homology and analysis of motif or domain showed that some genes may be important functional genes, such as those coding signaling proteins, proteins associated with ion channel, enzymes involved in hormone production, important transcription factors, translation initiation factors, etc. 7 of the novel full-length cDNAs are new isoforms produced by alternative splicing.


CONCLUSIONS
126 novel full-length cDNAs have been cloned from adrenal tissues by large-scale EST sequencing and bioinformatic tools in a rather short time, some of which may play an important role in regulating adrenal functions and tumorigenesis of adrenal gland.
Peng
Yongde
Y
Shanghai Institute of Endocrinology, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China.
Li
Yuebin
Y
Song
Huaidong
H
Gao
Guofeng
G
Huang
Changhui
C
Hu
Renming
R
Han
Zeguang
Z
Chen
Jialun
J
D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase / Q000235:genetics; D000311:Adrenal Glands / Q000502:physiology*; D000429:Alcohol Oxidoreductases / Q000235:genetics; D017398:Alternative Splicing; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D020869:Gene Expression Profiling; D020962:Heterotrimeric GTP-Binding Proteins / Q000235:genetics; D006801:Humans; D010673:Pheochromocytoma / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D054462:p21-Activated Kinases
D018076:DNA, Complementary; D000429:Alcohol Oxidoreductases; C019642:alcohol dehydrogenase (NADP+); D013741:3-Oxo-5-alpha-Steroid 4-Dehydrogenase; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases; D020962:Heterotrimeric GTP-Binding Proteins