title: A genome walking strategy for the identification of nucleotide sequences adjacent to known regions.
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.
authors.affiliation: College of Life Science, Hunan Normal University; State Key Laboratory Breeding Base of Microbial Molecular Biology, Changsha, People's Republic of China. hlw1225@hotmail.com
authors.forename: Hailong
authors.initials: H
authors.lastname: Wang
authors.forename: Ting
authors.initials: T
authors.lastname: Yao
authors.forename: Mei
authors.initials: M
authors.lastname: Cai
authors.forename: Xiuqing
authors.initials: X
authors.lastname: Xiao
authors.forename: Xuezhi
authors.initials: X
authors.lastname: Ding
authors.forename: Liqiu
authors.initials: L
authors.lastname: Xia
mesh_terms: D000490:Allium / Q000235:genetics; D016386:Chromosome Walking / Q000379:methods*; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D016930:Saccharopolyspora / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D004251:DNA Transposable Elements