Transcriptional Analysis of the DNA Polymerase Gene of Bombyx mori Parvo-like Virus (China Isolate)

다국어 초록 (Multilingual Abstract)

The Bombyx mori parvo-like virus (China isolate) DNA polymerase (BmDNV-3 dnapol) gene has been tentatively identified based on the presence of conserved motifs. In the present study, we perform a transcriptional analysis of the BmDNV-3 dnapol gene using the total RNA isolated from BmDNV-3 infected silkworm at different times. Northern blot analysis with a BmDNV-3 dnapol-specific riboprobe showed a major transcript of 3.3 kb. 5′-RACE revealed that the major transcription start point was located 20 nucleotides downstream of the TATA box. In a temporal expression analysis using differential RT-PCR, BmDNV-3 dnapol transcript was detected at low levels at 6 h.p.i., increased from 6 to 36 h.p.i., and remained fairly constant thereafter. Analysis of the predicted DNA polymerase sequence using neighborjoining and protein parsimony algorithms indicated that the predicted 1115-residue polypeptide contained five motifs associated with DNA polymerases synthetic activities and three additional motifs associated with polymerases possessing 3′to 5′exonuclease activity. The molecular phylogenetic analysis of this gene supported the placement of Bombyx mori parvo-like virus in a separate virus family.

참고문헌 (Reference)

1 Wu, "and regulation of a baculovirus DNA polymerase gene" 591-167 600,

2 Kim, E.K, "The terminal protein of a linear mitochondrial plasmid is encoded in the N-terminus of the DNA polymerase gene in white-rot fungus Pleurotus ostreatus" 38 : 283-290, 2000

3 Hermanns, "The linear mitochondrial plasmid pAL2-1 of a long-lived Podospora anserina mutant is an invertron encoding a DNA and RNA polymerase" 491-500, 1992

4 Chan, "The kalilo linear senescence-inducing plasmid of Neurospora is an invertron and encodes DNA and RNA polymerases" 225-237, 1991

5 Bernard, "The highy conserved amino acid sequence motif Tyr-Gly-Asp-Thr- Asp-Ser on alike DNA polymerase is required by a phase f29 DNA polymerase for protein-primed initiation and polymeri-zation" 4610-87 4614, 1990

6 Hess, "The complete nucleotide sequence of the egg drop syndrome virus An intermediate between mastadenoviruses and aviadenoviruses" 145-238 156,

7 Strasser, "The DNA polymerase gene from chlorella viruses PBCV-1 and NY-2A contains an intron with nuclear splicing sequences" 721-731, 1992

8 Thompson, J.D, "The CLUSTAL X windows interface: Flexible strategies for multiple sequence alignment aided by quality analysis tools" 25 : 4876-4882, 1997

9 Handa, H, "Structural features and expression analysis of a linear mitochondrial plasmid in rapeseed (Brasica napus L.)" 267 : 797-805, 2002

10 Klassen, R, "Structural and functional analysis of the killer element pPin1-3 from Pichia inositovora" 270 : 190-199, 2003

11 Lo, "Specific genomic fragment analysis of different geographical clinical samples of shrimp white spot syndrome virus" 175-185,

12 Vladimir, V, "Self-synthesizing DNA transposons in eukaryotes" 103 : 4540-4545, 2006

13 Lee, A.M, "Repetitive DNA insertion in a protein kinase ORF of a latent FSV (Feldmannia sp. virus) genome" 248 : 35-45, 1998

14 Blanco, L, "Relating structure to function in phi29 DNA polymerase" 271 : 8509-8512, 1996

15 Kozak, "Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes" 283-292, 1986

16 Ehrlich, "Physical characteristics in eukaryotic promoters" 4521-4540, 1983

17 Swofford, D.L., "PAUP*. Phylogenetic analysis using parsimony (*and other methods)" Sunderland, MA 1998

18 Wang, Y.J, "Organization and transcription strategy of genome of Bombyx mori bidensovirus (China isolate) VD1" 22 : 706-712, 2006

19 Sawada, "Nucleotide sequence of the adenovirus type 40 inverted terminal repeat Close relation to that of adenovirus type 5" ishi : 414-156 416, 1987

20 Lu, H.S, "Molecule biology of insect virus" China agriculture science and technology publishing company press 382-391, 1998

21 Bois, F, "Molecular cloning, sequence and expression of Aa-polB, a mitochondrial gene encoding a family B DNA polymerase from the edible basidiomycete Agrocybe aegerita" 261 : 508-513, 1999

22 Liu, J.J, "Identification, localization, transcription, and sequence analyses of the Choristoneura fumiferana nuclear polyhedrosis virus DNA polymerase gene" 209 : 538-549, 1995

23 Chen, L.L, "Identification of a nucleocapsid protein (VP35) gene of shrimp white spot syndrome virus and characterization of the motif important for targeting vp35 to the nuclei of transfected insect cells" 293 : 44-53, 2002

24 Bulach, D.M, "Group II nucleopolyhedrovirus subgroups revealed by a phylogenetic analysis of polyhedrin and DNA polymerase gene sequences" 73 : 59-73, 1999

25 Court, "Genetic organization and structural features of maranhar a senescence-inducing linear mitochondrial plasmid of Neurospora crassa" 385-397,

26 Verheust, C, "GIL16, a New Grampositive tectiviral phage related to the Bacillus thuringiensis GIL01 and the Bacillus cereus pBClin15 elements" 187 : 1966-1973, 2005

27 Morrison, "Eukaryotic DNA polymerase aminoacid sequence required for 3→5 exonuclease activity" sugi : 9473-88 9478,

28 Nakai, R, "Basidiomycetous fungus Flammulina velutipes harbors two linear mitochondrial plasmids encoding DNA and RNA polymerases" 190 : 99-102, 2000