Journal Article

Construction of Expression-ready cDNA Clones for KIAA Genes: Manual Curation of 330 KIAA cDNA Clones

Daisuke Nakajima, Noriko Okazaki, Hisashi Yamakawa, Reiko Kikuno, Osamu Ohara and Takahiro Nagase

in DNA Research

Published on behalf of Kazusa DNA Research Institute

Volume 9, issue 3, pages 99-106
Published in print January 2002 | ISSN: 1340-2838
Published online January 2002 | e-ISSN: 1756-1663 | DOI: http://dx.doi.org/10.1093/dnares/9.3.99

Show Summary Details

Preview

We have accumulated information on protein-coding sequences of uncharacterized human genes, which are known as KIAA genes, through cDNA sequencing. For comprehensive functional analysis of the KIAA genes, it is necessary to prepare a set of cDNA clones which direct the synthesis of functional KIAA gene products. However, since the KIAA cDNAs were derived from long mRNAs (> 4 kb), it was not expected that all of them were full-length. Thus, as the first step toward preparing these clones, we evaluated the integrity of protein-coding sequences of KIAA cDNA clones through comparison with homologous protein entries in the public database. As a result, 1141 KIAA cDNAs had at least one homologous entry in the database, and 619 of them (54%) were found to be truncated at the 5′ and/or 3′ ends. In this study, 290 KIAA cDNA clones were tailored to be full-length or have considerably longer sequences than the original clones by isolating additional cDNA clones and/or connected parts of additional cDNAs or PCR products of the missing portion to the original cDNA clone. Consequently, 265, 8, and 17 predicted CDSs of KIAA cDNA clones were increased in the amino-, carboxy-, and both terminal sequences, respectively. In addition, 40 cDNA clones were modified to remove spurious interruption of protein-coding sequences. The total length of the resultant extensions at amino- and carboxy-terminals of KIAA gene products reached 97,000 and 7216 amino acid residues, respectively, and various protein domains were found in these extended portions.

Keywords: large proteins; cDNA sequencing; manual curation; protein production

Journal Article.  0 words. 

Subjects: Genetics and Genomics

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.