博士 / 國立臺灣大學 / 動物學系 / 82 / Penaeus monodon-type baculovirus (MBV) was isolated and
purified from the hepatopancreata of MBV- infected Penaeus
monodon Fabricius. MBV DNA was extracted and used as a
template in polymerase chain reaction (PCR). The primers were
chosen from conserved regions of the polyhedrin gene of
Autographa califor- nica nuclear polyhedrosis virus (AcNPV).
One DNA fragment (674 base pairs) was amplified after PCR.
There is 65% homology between the predicted amino acid sequence
of this PCR product with that of polyhedrin polypeptide of
AcNPV. Nucleotide sequence analysis indicates that the
amplified DNA is the partial open reading frame of the MBV
polyhedrin gene. This 674 bp DNA fragment was subsequently
used as a probe in a dot blot analysis; the result showed that
the probe is able to hybridize to the DNA extracted from the
purified MBV and from the MBV- infected P. monodon, but not
from the MBV-uninfected P. monodon. A digoxigenin-labeled DNA
probe was used for in situ detection of the Penaeus monodon
Fabricius baculovirus (MBV) genome in cultured prawns. We
verified the specificity of probe with dot blot analysis, it
suggested the cloned MBV polyhedrin DNA or MBV polyhedrin PCR
product can be used as a MBV-specific probe. This probe was
further applied for detection MBV using in situ hybri- dization
method and MBV DNA was observed in the hepatopancreata, midgut,
lymphoid organ, and gills. Subsequently, we investigated the
susceptibility to MBV infection of various life-stages of the
prawn. The MBV genome was seen in the zoea, mysis and
postlarval stage, whereas MBV DNA was not detected in eggs and
nauplii. In additon, we quantified the results from in situ
hybridization with a image analyzer and compared the cell
infected rate of cultured P. monodon collected from various
farms in Taiwan.
| Identifer | oai:union.ndltd.org:TW/082NTU00312007 |
| Date | January 1994 |
| Creators | Lu, Chow-Chin, 盧秀琴 |
| Contributors | Kou, Guang-Hsiung; Chen, Shiu-Nan, 郭光雄；陳秀男 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 164 |
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