A genome walking strategy for the identification of nucleotide sequences adjacent to known regions

作者：Wang, Hailong; Yao, Ting; Cai, Mei; Xiao, Xiuqing; Ding, Xuezhi; Xia, Liqiu^*

来源：Biotechnology Letters, 2013, 35(2): 279-284.

DOI：10.1007/s10529-012-1076-3

• Cloning
• Genome walking
• Single primer PCR
• Transposon

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.

• 出版日期2013-2
• 单位湖南师范大学