• The Plant Pathology Journal
• /
• v.21 no.1
• /
• pp.13-20
• /
• 2005

Chlorella viruses are large icosahedral, plaque-forming, dsDNA viruses that infect certain unicellular, chlorellalike green algae. The genomic DNA of over 300 kb contains many useful genes and promoters. Over 40 chlorella viruses have been isolated from fresh water in Korea since 1998. The viruses were amplified initially in chlorella strain NC64A, and pure isolates were obtained by repeated plaque isolation. SDS-PAGE analysis revealed similar but distinct protein patterns, both among the group of purified viruses and in comparison with the prototype chlorella virus PBCV-1. Digestions of the 330- to 350-kb genomic DNAs with 10 restriction enzymes revealed different restriction fragment patterns among the isolates. The tRNA-coding regions of 8 chlorella viruses were cloned and sequenced. These viruses contain 14-16 tRNA genes within a 1.2- to 2-kb region, except for the SS-1 isolate, which has a 1039-bp spacer in a cluster of 11 tRNA genes. Promoter regions of several early genes were isolated and their activities were analyzed in transformed chlorella. Some promoters showed stronger activity than commonly used CaMV 35S promoter and chlorella transformation vectors for heterologous protein are beings constructed using these promoters.

• The Plant Pathology Journal
• /
• v.28 no.4
• /
• pp.433-438
• /
• 2012

Dinoflagellates are considered one of the most abundant and diverse groups of marine microplankton and viruses are recognized as one of the significant factors affecting the plankton dynamics. Here, we report basic characteristics of a new dinoflagellate-infecting virus, Heterocapsa pygmaea DNA virus (HpygDNAV) which infects a toxic dinoflagellate, H. pygmaea. HpygDNAV is a polyhedral large virus (ca. 160-170 nm in diameter) propagating in its host's cytoplasm. Because of the virion size, appearance in thin sections, and propagation characteristics, HpygDNAV is assumed to harbor a large double-stranded DNA genome; i.e., HpygDNAV is most likely a nucleocytoplasmic large DNA virus (NCLDV) belonging to the family Phycodnaviridae. Its infectivity is strain-specific, rather than species-specific, as is the case for other algal viruses. The burst size and latent period are estimated to be roughly 100-250 infectious units $cell^{-1}$ and < 96 h, respectively.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.6
• /
• pp.952-960
• /
• 2006

As a unicellular green alga that possesses many of the metabolic pathways present in higher plants, Chlorelia offers many advantages for expression of heterologous proteins. Since strong and constitutive promoters are necessary for efficient expression in heterologous expression systems, the development of such promoters for use in the Chlorella system was the aim of this study. Proteins encoded by the early genes of algal viruses are expressed before viral replication, probably by the host transcriptional machinery, and the promoters of these genes might be useful for heterologous expression in Chlorella. In this study, putative promoter regions of DNA polymerase, ATP-dependent DNA ligase, and chitinase genes were amplified from eight Korean Chlorella virus isolates by using primer sets designed based on the sequence of the genome of PBCV-1, the prototype of the Phycodnaviridae. These putative promoter regions were found to contain several cis-acting elements for transcription factors, including the TATA, CAAT, NTBBF1, GATA, and CCAAT boxes. The amplified promoter regions were placed into Chlorella transformation vectors containing a green fluorescence protein (GFP) reporter gene and the Sh ble gene for phleomycin resistance. C. vulgaris protoplasts were transformed and then selected with phleomycin. The GFP fluorescence intensities of cells transformed with chitinase, DNA polymerase, and DNA ligase gene promoter-GFP fusion constructs were 101.5, 100.8, and 95.8%, respectively, of that of CaMV 35S-GFP-transformed Chlorella cells. These results demonstrate that these viral promoters are active in transformed Chlorella.