Single nucleotide polymorphism discovery of Pinus radiata with chromosome walking PCR method |
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| Authors: | Wei Li Hui Li Xiaoyang Chen Harry Wu |
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| Institution: | (1) College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, China;(2) CSIRO Forest Bioscience, Canberra, 2600, Australia;(3) College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing, 100083, China;(4) College of Forestry, South China Agricultural University, Guangzhou, 200064, China;(5) CSIRO Forest Bioscience, Canberra, 2600, Australia |
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| Abstract: | In this paper, the basic principle of chromosome walking is presented and we used an actin gene of radiata pine (Pinus radiata) as an example to conduct upstream and downstream chromosome walking for EST sequences. The full genomic sequence (2154 bp)
of the actin gene, including promoters 5′ UTR, CDS and 3′ UTR, was identified by chromosome walking. PCR amplification and DNA band sequencing
from 200 unrelated radiata pine trees revealed a total of 21 SNPs for the actin gene, three in the promoter region, 15 in CDS and 4 in 3′ UTR. The results of this experiment provide a technical framework
for SNPs discovery in none coding regions of candidate genes.
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Translated from Acta Botanica Boreali-Occidentalia Sinica, 2007, 27 (8): 1571–1576 译自: 西北植物学报] |
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| Keywords: | radiata pine single nucleotide polymorphism (SNP) chromosome walking none coding region |
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