[Cloning and identification of telomere and internal origin of the endophytic Streptomyces linear plasmid pYY8L from Allium fistulosum].
Strain 36R-2-1B was isolated from Allium fistulosum and identified as Streptomyces spp., harboring an approximate 280 kb linear plasmid designated pYY8L.


OBJECTIVE
Cloning, sequencing and analysis of telomere and internal replication origin of pYY8L.


METHODS
pYY8L telomere was cloned by a modified procedure--"alkaline treatment and enzyme digestion in gel". The internal replication origin of pYY8L was cloned by construction of a cosmid library and then subcloning.


RESULTS
An approximate 280 kb DNA band (pYY8L) of strain 36R-2-1B was detected by pulsed-field gel electrophoresis. The 152-bp telomere, containing six small palindromes and potentially forming complicated secondary structure, was cloned and sequenced. The replication origin of pYY8L was initially cloned on a cosmid and then sub-cloned as a 4891-bp autonomous replication sequence. This sequence was predicted to contain six genes, two of them resembling replication genes of Streptomyces linear plasmid SCP1, but their adjacent iterons were different.


CONCLUSIONS
New telomere and internal replication origin of Streptomyces linear plasmid pYY8L were cloned and identified. This is the first example of report on telomere and replication genes of linear plasmid from endophytic Streptomyces.
Yang
Yong
Y
Key laboratory of Synthetic Biology, Shanghai Institute of Plant Physiology and Ecology, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China. yyang@sibs.ac.cn
Zhong
Li
L
Tian
Xinli
X
Cheng
Qiuxiang
Q
Chen
Zhenhua
Z
Qin
Zhongjun
Z
D000490:Allium / Q000235:genetics*; D003001:Cloning, Molecular; D003227:Conjugation, Genetic / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D018741:Replication Origin / Q000235:genetics*; D013302:Streptomyces / Q000235:genetics*; D016615:Telomere / Q000235:genetics* / Q000378:metabolism
D004269:DNA, Bacterial