Abstract
To identify the transposon insertion sites in a soil actinomycete, Saccharopolyspora spinosa, a genome walking approach, termed SPTA-PCR, was developed. In SPTA-PCR, a simple procedure consisting of TA cloning and a high stringency PCR, following the single primer-mediated, randomly-primed PCR, can eliminate non-target DNA fragments and obtain target fragments specifically. Using SPTA-PCR, the DNA sequence adjacent to the highly conserved region of lectin coding gene in onion plant, Allium chinense, was also cloned.
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (31070006), the National Basic Research Program (973) of China (2012CB722300), "973" special preliminary study program (2011CB311800), and the National High Technology Research and Development program (863) of China (2011AA10A203).
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Wang, H., Yao, T., Cai, M. et al. A genome walking strategy for the identification of nucleotide sequences adjacent to known regions. Biotechnol Lett 35, 279–284 (2013). https://doi.org/10.1007/s10529-012-1076-3
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DOI: https://doi.org/10.1007/s10529-012-1076-3