TY - JOUR
T1 - Characterization of a novel plasmid from extremely halophilic Archaea
T2 - Nucleotide sequence and function analysis
AU - Ye, Xuecheng
AU - Ou, Jianhong
AU - Ni, Lina
AU - Shi, Wanliang
AU - Shen, Ping
N1 - Funding Information:
We thank Dr. W.F. Doolittle for kindly providing the pWL102 plasmid. We are grateful to Dr. Merck and Dr. Sharp and Dohme Research Lab. for providing mevinolin. This work was supported by the National Science Foundation of China (Nos. 39070009, 39770009).
PY - 2003/4/11
Y1 - 2003/4/11
N2 - We determined the complete nucleotide sequence of the 16 341 bp plasmid pHH205 of the extremely halophilic archaeon Halobacterium salinarum J7. The plasmid has a G+C content of 61.1%. A number of direct and inverted repeat sequences were found in pHH205, while no insertion sequences were found. Thirty-eight large open reading frames (ORFs) were identified in both strands, and most of them had no significant similarities to known proteins. A putative protein encoded by ORF31 showed 20-41% homology to some hypothetical proteins, which are annotated in several archaeal genome databases as predicted nucleic acid-binding proteins containing PIN domain. Sequence analysis using the GC skew procedure predicted a possible origin of replication. A 4.8 kb PvuII-SnaBI fragment containing both this region and ORF31 was shown to be able to restore replicate of pWL102, a replicon-deficient plasmid in Haloferax volcanii and in H. salinarum R1. Several methods failed to completely cure H. salinarum J7 of pHH205, suggesting that the plasmid probably played an important role in the growth and metabolism of the host. Our work describes a novel haloarchaeal replicon, which may be useful in the construction of cloning and shuttle vectors.
AB - We determined the complete nucleotide sequence of the 16 341 bp plasmid pHH205 of the extremely halophilic archaeon Halobacterium salinarum J7. The plasmid has a G+C content of 61.1%. A number of direct and inverted repeat sequences were found in pHH205, while no insertion sequences were found. Thirty-eight large open reading frames (ORFs) were identified in both strands, and most of them had no significant similarities to known proteins. A putative protein encoded by ORF31 showed 20-41% homology to some hypothetical proteins, which are annotated in several archaeal genome databases as predicted nucleic acid-binding proteins containing PIN domain. Sequence analysis using the GC skew procedure predicted a possible origin of replication. A 4.8 kb PvuII-SnaBI fragment containing both this region and ORF31 was shown to be able to restore replicate of pWL102, a replicon-deficient plasmid in Haloferax volcanii and in H. salinarum R1. Several methods failed to completely cure H. salinarum J7 of pHH205, suggesting that the plasmid probably played an important role in the growth and metabolism of the host. Our work describes a novel haloarchaeal replicon, which may be useful in the construction of cloning and shuttle vectors.
KW - Curing
KW - Halobacterium salinarum
KW - Plasmid pHH205
KW - Sequence and function analysis
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U2 - 10.1016/S0378-1097(03)00175-7
DO - 10.1016/S0378-1097(03)00175-7
M3 - Article
C2 - 12694910
AN - SCOPUS:0037432696
SN - 0378-1097
VL - 221
SP - 53
EP - 57
JO - FEMS microbiology letters
JF - FEMS microbiology letters
IS - 1
ER -