Use of chromosome walking in discovery of single-nucleotide polymorphism in noncoding regions of a candidate actin gene in Pinus radiata

• Autorzy: Li W. , Li H. , Wu H. , Chen X.-Y.
• Języki publikacji: EN

Abstrakty

EN

Untranslated regions (UTRs) of eukaryotic mRNAs plav crucial roles in post-transcriptional regulation of gene expression via the modulation of nucleocytoplasmic mRNA transport, translation efficiency, subcellular localization, and message stability. Single-nucleotide polymorphisms (SNPs) in UTRs of a candidate gene may also change the post-transcriptional regulation of a gene or function by nucleotide mutation. For species that have not been entirely sequenced genomically, new methods need to be devised to discover SNPs in noncoding regions of candidate genes. In this study, based on the expressed sequence tag (EST) of Pinus radiata (Monterey pine), we obtained all the sequences of UTRs of the actin gene by using a chromosome walking method. We also detected all the SNPs in and around the coding region of the actin gene. In this way, the full genomie sequence (2154 bp) of the actin gene was identified, including the 5'UTR, introns, the coding sequence, and the 3'UTR. PCR amplification and DNA fragment sequencing from 200 unrelated P. radiata trees revealed a total of 21 SNPs in the actin gene, of which 3 were located in the 5'UTR, 3 in the introns, 10 in the coding sequence, and 5 in the 3'UTR. We show that chromosome walking can be used for obtaining the sequence of UTRs, and then, based on this sequence, to discover SNPs in the noncoding regions of candidate genes from this species without an entire genomic sequence.

Słowa kluczowe

EN

chromosome; single-nucleotide polymorphism; non-coding region; candidate gene; Pinus radiata; detection; gene expression; nucleotide mutation; genome sequence

• Wydawca: -
• Czasopismo: Journal of Applied Genetics
• Rocznik: 2010
• Tom: 51
• Numer: 3
• Opis fizyczny: p.275-281,fig.,ref.

Twórcy

autor

Li W.

• Key Laboratory for Genetics and Breeding of Forest Trees and Ornamental Plants, Ministry of Education, National Engineering Laboratory for Forest Tree Breeding, Beijing Forestry University, Beijing, China
• CSIRO Plant Industry, Canberra, Australia

autor

Li H.

• Key Laboratory for Genetics and Breeding of Forest Trees and Ornamental Plants, Ministry of Education, National Engineering Laboratory for Forest Tree Breeding, Beijing Forestry University, Beijing, China

autor

Wu H.

• CSIRO Plant Industry, Canberra, Australia

autor

Chen X.-Y.

• College of Forestry, South China Agriculture University, Guangzhou, China

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