|Genome Sequencing Section
DOE Human Genome Program Contractor-Grantee Workshop
sequence analysis of 918 human cDNA clones harboring long and nearly full-length
Nobuo Nomura, Takahiro Nagase, Ken-ichi Ishikawa, Reiko Kikuno,
Mikita Suyama, Nobuyuki Miyajima, Ayako Tanaka, Hirokazu Kotani, and Osamu
One of the goals of the Kazusa human cDNA project is to accumulate and exploit information on coding sequences of unidentified human cDNA clones harboring long and nearly full-length inserts. We have so far determined the entire sequences of 918 clones (KIAA0001-KIAA0918) with average size of 5.0kb. Among the clones, 268 were obtained from human immature myeloid cell like KG-1 and 650 from human brain. All the KG-1 and 25 brain cDNA clones were selected under the criteria that the clones carry inserts with at least 90% of the length of the corresponding transcripts. As another novel approach, 588 brain clones were selected based upon their capabilities to produce proteins in vitro with molecular weight larger than 50kDa. Since approximately 50% of the cDNA clones isolated by either method was found to retain an in-frame termination codon upstream of the first ATG codon, it was speculated that more than half of the clones analyzed harbored complete ORF. And we concluded that clones with complete ORF can be selected efficiently by either of the procedures. It turned out that 750 out of 918 clones encoded proteins larger than 50kDa. By computer analysis, we successfully assigned two thirds of 750 clones to the functional categories such as genes for cell signalling/communication (198 clones, 26.3%), cell structure/motility (106 clones, 14.2%), nucleic acid managing (112 clones, 15%), protein managing (30 clones, 4%), metabolism (17 clones, 2.3%), and cell division (9 clones, 1.2 %) Database search also revealed that most of ESTs currently registered fell in the region within 2kb from the 3'-end of our cDNA sequences. When the expression profiles of the cDNA clones were examined among over a dozen human tissues, approximately 80% of the clones from KG-1 and 20% of the clones from brain were expressed ubiquitously. The chromosomal locations of these clones were also determined.
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