Journal of Microbiology

Hee Jin Shim

2 Articles

Cloning and Molecular Characterization of a Novel Rolling-Circle Replicating Plasmid, pK1S-1, from Bacillus thuringiensis subsp. kurstaki K1: Ming Shun Li , Jong Yul Roh , Xueying Tao , Zi Niu Yu , Zi Duo Liu , Qin Liu , Hong Guang Xu , Hee Jin Shim , Yang-Su Kim , Yong Wang , Jae Young Choi , Yeon Ho Je; J. Microbiol. 2009;47(4):466-472. Published online September 9, 2009; DOI: https://doi.org/10.1007/s12275-009-0020-2

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Bacillus thuringiensis, an entomopathogenic bacterium belonging to the B. cereus group, harbors numerous extra-chromosomal DNA molecules whose sizes range from 2 to 250 kb. In this study, we used a plasmid capture system (PCS) to clone three small plasmids from B. thuringiensis subsp. kurstaki K1 which were not found in B. thuringiensis subsp. kurstaki HD-1, and determined the complete nucleotide sequence of plasmid pK1S-1 (5.5 kb). Of the six putative open reading frames (ORF2-ORF7) in pK1S-1, ORF2 (MobK1) showed approximately 90% aa identity with the Mob-proteins of pGI2 and pTX14-2, which are rolling circle replicating group VII (RCR group VII) plasmids from B. thuringiensis. In addition, a putative origin of transfer (oriT) showed 95.8% identity with those of pGI2 and pTX14-2. ORF3 (RepK1) showed relatively low aa identity (17.8~25.2%) with the Rep protein coded by RCR plasmids, however. The putative double- strand origin of replication (dso) and single-strand origin of replication (sso) of pK1S-1 exhibited approximately 70% and 64% identities with those of pGI2 and pTX14-2. ORF6 and 7 showed greater than 50% similarities with alkaline serine protease, which belongs to the subtilase family. The other 2 ORFs were identified as hypothetical proteins. To determine the replicon of pK1S-1, seven subclones were contructed in the B. thuringiensis ori-negative pHT1K vector and were electroporated into a plasmid cured B. thuringiensis strain. The 1.6 kb region that included the putative ORF3 (Rep1K), dso and ORF4, exhibited replication ability. These findings identified pK1S-1 as a new RCR group VII plasmid, and determined its replication region.

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Isolation and Characterization of a Lymantria dispar Multinucleocapsid Nucleopolyhedrovirus Isolate in Korea: Hee Jin Shim , Jong Yul Roh , Jae Young Choi , Ming Shun Li , Soo Dong Woo , Hyun Woo Oh , Kyung Saeng Boo , Yeon Ho Je; J. Microbiol. 2003;41(4):306-311.

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Abstract PDF: In Korea, a Lymantria dispar multinucleocapsid nucleopolyhedrovirus, LdMNPV-NM, was isolated and characterized from dead L. dispar larvae. The polyhedra of LdMNPV-NM were irregularly shaped with a diameter of 1.62?0.33 ?m. Numerous virions comprised of the multinucleocapsid were evident in the electron microscopic examination of the polyhedra cross sections. These polyhedra were composed of a major protein of 30 kDa. The restriction enzyme digestion patterns of LdMNPV-NM showed that this isolate had some different fragments from those of the Gypchek팜?LdMNPV isolate, although their overall profiles were similar. The deduced amino acid sequence of the enhancin gene of LdMNPV-NM showed differences when compared to previously reported enhancin genes of other LdMNPV strains. These results suggested that the LdMNPV-NM isolate from Korea was a new NPV strain and had a new enhancin gene.

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