© 2002 by Kazusa DNA Research Institute
Construction of Expression-ready cDNA Clones for KIAA Genes: Manual Curation of 330 KIAA cDNA Clones
Kazusa DNA Research Institute 1532-3 Yana, Kisarazu, Chiba 292-0812, Japan
* To whom correspondence should be addressed. Tel. +81-438-52-3930, Fax. +81-438-52-3931, E-mail: nagase{at}kazusa.or.jp
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.
Key words: large proteins; cDNA sequencing; manual curation; protein production
![]()
CiteULike
Connotea
Del.icio.us What's this?
This article has been cited by other articles:
![]() |
V. Patel, B. L. Hood, A. A. Molinolo, N. H. Lee, T. P. Conrads, J. C. Braisted, D. B. Krizman, T. D. Veenstra, and J. S. Gutkind Proteomic Analysis of Laser-Captured Paraffin-Embedded Tissues: A Molecular Portrait of Head and Neck Cancer Progression Clin. Cancer Res., February 15, 2008; 14(4): 1002 - 1014. [Abstract] [Full Text] [PDF] |
||||
![]() |
H. Ueda, R. Nagae, M. Kozawa, R. Morishita, S. Kimura, T. Nagase, O. Ohara, S. Yoshida, and T. Asano Heterotrimeric G Protein {gamma} Subunits Stimulate FLJ00018, a Guanine Nucleotide Exchange Factor for Rac1 and Cdc42 J. Biol. Chem., January 25, 2008; 283(4): 1946 - 1953. [Abstract] [Full Text] [PDF] |
||||
![]() |
D. Bhattacharyya and B. S. Glick Two Mammalian Sec16 Homologues Have Nonredundant Functions in Endoplasmic Reticulum (ER) Export and Transitional ER Organization Mol. Biol. Cell, March 1, 2007; 18(3): 839 - 849. [Abstract] [Full Text] [PDF] |
||||
![]() |
T. Nagase, H. Koga, and O. Ohara Kazusa mammalian cDNA resources: towards functional characterization of KIAA gene products Brief Funct Genomic Proteomic, March 1, 2006; 5(1): 4 - 7. |
||||
![]() |
D. Nakajima, K. Saito, H. Yamakawa, R. F. Kikuno, M. Nakayama, R. Ohara, N. Okazaki, H. Koga, T. Nagase, and O. Ohara Preparation of a Set of Expression-Ready Clones of Mammalian Long cDNAs Encoding Large Proteins by the ORF Trap Cloning Method. DNA Res, January 1, 2005; 12(4): 257 - 267. [Abstract] [Full Text] [PDF] |
||||
![]() |
M. Murakami, K. Shimada, M. Kawai, and H. Koga InCeP: Intracellular Pathway Based on mKIAA Protein-Protein Interactions DNA Res, January 1, 2005; 12(5): 379 - 387. [Abstract] [Full Text] [PDF] |
||||
![]() |
The Ludwig-FAPESP Transcript Finishing Initiative, M. C. Sogayar, and A. A. Camargo A Transcript Finishing Initiative for Closing Gaps in the Human Transcriptome Genome Res., July 1, 2004; 14(7): 1413 - 1423. [Abstract] [Full Text] [PDF] |
||||
![]() |
R. Kikuno, T. Nagase, M. Nakayama, H. Koga, N. Okazaki, D. Nakajima, and O. Ohara HUGE: a database for human KIAA proteins, a 2004 update integrating HUGEppi and ROUGE Nucleic Acids Res., January 1, 2004; 32(90001): D502 - 504. [Abstract] [Full Text] [PDF] |
||||






