TRANSCRIPTOME 2002: From Functional Genomics to Systems
Prepare a Set of Human cDNA Clones Encoding Large Proteins in an Expression-Ready Form
Takahiro Nagase, Noriko Okazaki, Daisuke Nakajima, Hisashi Yamakawa, Reiko Kikuno, and Osamu Ohara, Kazusa DNA Research Institute/Department of Human Gene Research, Kisarazu, Chiba, JAPAN
We have focused our sequencing efforts on long cDNAs (>4 kb) encoding relatively large proteins (>500 a.a.) from brain. The total number of cDNAs entirely sequenced (KIAA plus some of FLJ cDNAs) has exceeded 2000. Besides these cDNA clones, we have also accumulated more than 1500 cDNA clones coding for large proteins first identified by other groups. However, the coding sequences of some cloned cDNAs were predicted to be truncated due to incompleteness of the cDNAs or spurious interruption by intron(s) and/or reverse transcription error(s). Although we have continuously updated our cDNA sequences through experimentally revising these spurious coding interruption as far as possible, our cDNA clones cannot always produce the authentic proteins because these revisions have been made only in sequence information. Thus, preparation of a set of expression-ready cDNA clones encoding large proteins becomes our urgent mission because this must play a critical role for comprehensive functional analysis of human genes. We have conducted manual curation of our cDNA clones one by one to yield cDNA clones which can produce authentic KIAA proteins by combination of currently available methods. Because large cDNAs are generally difficult targets to obtain in an expression-ready form, our efforts will provide an invaluable human gene resource to the research community.
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