Background
Methods
Algal strains and culture conditions
Detection of viruses
Plaque assay and virus isolation
Electric microscopic observation
SDS-PAGE analysis
N-terminal amino acid sequence analysis and amino acid sequence homology search
Pulsed-field gel electrophoresis (PFGE)
Extraction of CvV-BW1 DNA
Digestion of CvV-BW1 DNA with restriction enzymes
HPLC analysis of methylated nucleotides
Hyaluronan labeling
DNA polymerase gene analyses
Results and discussion
Ecological studies of viruses in Lake Biwa
Sampling date | Water temp. (°C) | Site 1 | Site 2 | Site 3 | Site 4 | Site 5 | Site 6 | Site 7 | Site 8 | Site 9 |
---|---|---|---|---|---|---|---|---|---|---|
2004 May | -- | 2.67 | 21.3 | 21.3 | 2.67 | 21.3 | 2.67 | ND | 85.3 | 21.3 |
June | 16.5-19.5 | 21.3 | 5.33 | 10.7 | 5.33 | 5.33 | -- | 10.7 | 171.0 | 42.7 |
July | 29.3-32.0 | ND | 0.67 | 0.67 | ND | 1.33 | -- | ND | 10.7 | 0.67 |
Sept. | 24.0-25.5 | 0.67 | 5.33 | 10.7 | 10.7 | -- | -- | -- | -- | -- |
Oct. | 16.0-16.9 | 0.67 | 1.33 | 10.7 | 5.33 | -- | -- | -- | -- | -- |
Nov. | 10.2-12.0 | 5.33 | 5.33 | 5.33 | 5.33 | -- | -- | -- | -- | -- |
Dec. | 8.8-11.2 | 0.67 | 1.33 | 21.3 | 0.33 | -- | -- | -- | -- | -- |
2005 Jan. | 3.9-6.8 | 21.3 | 1.33 | 0.67 | 0.67 | -- | -- | -- | -- | -- |
Feb. | 5.2-8.3 | 1.33 | 1.33 | 10.7 | 5.33 | -- | -- | -- | -- | -- |
Mar. | 8.0-9.4 | 5.33 | 10.7 | 21.3 | 5.33 | -- | -- | -- | -- | -- |
Apr. | 17.0-18.8 | 5.33 | 21.3 | 21.3 | 5.33 | -- | -- | -- | -- | -- |
June | 23.0-24.0 | 5.33 | 10.7 | 42.7 | 10.7 | -- | -- | -- | -- | -- |
Plaque-forming assay
Protein of CvV-BW1
Size of CvV-BW1 DNA
Resistance/susceptibility of CvV-BW1 DNA to restriction enzymes
Restriction enzyme | Recognition sequence |
---|---|
Bal I | TGGCCA |
Bam HI | GGATCC |
Eco RI | GAATTC |
Hae III | RGCGCY |
Hin dIII | AAGCTT |
Hpa I | GTTAAC |
Nco I | CCATGG |
Not I | GCGGCCGC |
Pst I | CTGCAG |
Pvu II | CAGCTG |
Sac I | GAGCTC |
Sal I | GTCGAC |
Sca I | AGTACT |
Sfi I | GGCCNNNNNGGCC |
Sse 8387I | CCTGCAGG |
Restriction enzyme | Recognition sequence |
---|---|
Bgl II | AGATCT |
Dra I | TTTAAA |
Eco RV | GATATC |
Nde I | CATATG |
Mss I | GTTTAAAC |
Sau 3AI | GATC |
Ssp I | ACTAGT |
Swa I | ATTTAAAT |
Xba I | TCTAGA |