Abstract
A recombination-based strategy was developed for the assemble of valid open-reading frames. Compared with the conventional reverse transcription-polymerase chain reaction (PCR) approach, this strategy eliminates the inaccurate reverse transcription step but uses the efficient yeast recombination system to assemble the PCR-amplified exons of a gene. This strategy is particularly useful for the construction of full-length open-reading frames disrupted by a small number of introns. The number of valid introns in 862 rice kinase-coding genes was analyzed.
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Abbreviations
- ORF:
-
open-reading frame
- RT-PCR:
-
reverse transcription-polymerase chain reaction
References
Arabidopsis Genome Initiative (2000) Analysis of the genome sequence of the flowering plantArabidopsis thaliana. Nature 408: 796–815.
Chen DH and Ronald PC (1999) A rapid DNA minipreparation method suitable for AFLP and other PCR applications. Plant Mol Biol Rep 17: 53–57.
Chen X, Ding X, and Song WY (2003) Isolation of plasmid DNA rescued from single colonies ofAgrobacterium tumefaciens by means of rolling circle amplification. Plant Mol Biol Rep 21: 411–415.
Chevray PM and Nathans D (1992) Protein interaction cloning in yeast: identification of mammalian proteins that react with the leucine zipper of Jun. Proc Natl Acad Sci USA 89: 5789–5793.
Ding X, Snyder AK, Shaw R, Farmerie WG, and Song WY (2003) Direct re-transformation of yeast with plasmid DNA isolated from single yeast colonies using rolling circle amplification. Biotechniques 35: 774–779.
Gera JF, Hazbun TR, and Fields S (2002) Array-based methods for identifying protein-protein and protein-nucleic acid interactions. Methods Enzymol 350: 499–510.
Kikuchi S, Satoh K, Nagata T, Kawagashira N, Doi K, Kishimoto N, Yazaki J, Ishikawa M, Yamada H, Ooka H, et al. (2003) Collection, mapping, and annotation of over 28,000 cDNA clones from japonica rice. Science 301: 376–379.
Roberts JD, Bebenek K, and Kunkel TA (1988) The accuracy of reverse transcriptase from HIV-1. Science 242: 1171–1173.
Yamada K, Lim J, Dale JM, Chen H, Shinn P, Palm CJ, Southwick AM, Wu HC, Kim C, Nguyen M, et al. (2003) Empirical analysis of transcriptional activity in theArabidopsis genome. Science 302: 842–846.
Yu D, Ellis HM, Lee EC, Jenkins NA, Copeland NG, and Court DL (2000) An efficient recombination system for chromosome engineering inEscherichia coli. Proc Natl Acad Sci USA 97: 5978–5983.
Yuan Q, Ouyang S, Wang A, Zhu W, Maiti R, Lin H, Hamilton J, Haas B, Sultana R, Cheung F, et al. (2005) The institute for genomic research Osal rice genome annotation database. Plant Physiol 138: 18–26.
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Chen, X., Tao, Y., Liu, GZ. et al. A recombination-based strategy for assembling open-reading frames. Plant Mol Biol Rep 23, 397–403 (2005). https://doi.org/10.1007/BF02788887
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DOI: https://doi.org/10.1007/BF02788887