[논문]Isolation and Characterization of Chlorella Virus from Fresh Water in Korea and Application in Chlorella Transformation System

Park, Hye-Jin; Yoon, Hong-Mook; Jung, Heoy-Kyung; Choi, Tae-Jin

doi:10.5423/ppj.2005.21.1.013

Isolation and Characterization of Chlorella Virus from Fresh Water in Korea and Application in Chlorella Transformation System 원문보기

The plant pathology journal, v.21 no.1, 2005년, pp.13 - 20

Park, Hye-Jin (Department of Microbiology, Pukyong National University) , Yoon, Hong-Mook (Department of Microbiology, Pukyong National University) , Jung, Heoy-Kyung (Department of Microbiology, Pukyong National University) , Choi, Tae-Jin (Department of Microbiology, Pukyong National University)

Abstract ▼ AI-Helper

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.

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가설 설정

The relative location and orientations of primers used for PCR and sequencing. B. The arrangement of the tRNA genes of selected viiuses that were isolated in Korea.
Cells at the initial stage of virus infection. B. Vims particles have accumulated in the cytoplasm. C.

제안 방법

2. Restriction fragment length polymorphism analysis of Chlorella virus genomic DNAs that were digested with BmHI lane M, DNA size marker; lane 1, PBCV-1; lane 2, DJ; lane 3, HS-1; lane 4, HS-2; lane 5, YK; lane 6, SS-1; lane 7, SS-2; lane 8, KH-1; lane 9, KH-2.

후속연구

, 1995). Therefore, differences in the structural protein profiles may or may not reflect molecular diversity, and further characterization of the genomic DNA is necessary for the detailed 이assification of these viruses.

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