Compositions and methods for inhibiting white spot syndrome virus (WSSV) infection
A technology for white spot syndrome and viruses, applied in biochemical equipment and methods, chemical instruments and methods, viruses, etc., can solve undetermined problems
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Embodiment 1
Example 1. Expression of viral proteins and protein fragments
[0090] Four major nucleocapsid and envelope proteins from WSSV were evaluated. The primary and secondary structural motifs of each protein were modeled using the MacVector software package. Predictions of secondary structural features based on the amino acid sequence of each protein were tested using multiple predictive algorithms. The results from each prediction method were averaged and this information was used, along with additional predictive information on hydrophilicity, surface probability, flexibility and antigenicity index, to select each protein fraction to be expressed in the fusion system. Portions of viral proteins that may potentially interact with cellular receptors in the viral host may be exposed on the surface of the protein. Alternatively, the interactive parts of each protein may be contained within a single domain. By using the predictive information, likely portions of each viral protein t...
Embodiment 2
Example 2. Inhibition of WSSV infection using Vp28 protein fragments
[0093] Inhibition of WSSV infection using Vp28 fragments was performed as follows. A total of twelve 9 liter plastic aquariums (31 ppt salinity 30°C) were used to house the animals from the time the animals were received to the end of the experiment. The boxes are distributed randomly on two separate rack systems, each with its own water circulation system. In addition to the test group, two sentinel tanks and two positive control tanks were used to monitor possible pathogen escape from the exposure tank and to determine the virulence of the virus, respectively.
[0094] Six experimental feeds were prepared for bioassays. Two viral fusion proteins, one containing the Vp28 fragment and the other containing the Vp35 fragment, were used alone or in combination at two different concentrations to prepare the extruded feed. Immature vannamei (Penaeus vannamei) were fed the experimental diet for 72 hours prior t...
Embodiment 3
Example 3. White spot syndrome virus challenge
[0098] Pacific white shrimp (Peneaus vannamei, average weight 5 g) were grouped and housed in 9 liter flow boxes on an Aquatic Habitats racksystem. 4-8 animals per box, 3 boxes per group. The artificial sea salt was dissolved in Nano pure distilled water to a final salinity of 28ppt and kept at 28°C. Shrimp were housed in 9 boxes and fed one of 3 different diets. The control diet was Zeigler Brothers SI-35 grow-out diet. Both experimental diets were prepared in the laboratory using ground SI-35 as a matrix. The IgY feed had anti-Vp28 IgY added at 0.1%. Vp28 feed was prepared by adding 40 ml / kg of crude broth from CP Kelco runAB04903 (estimated Vp28 fusion concentration 10-40 g / ton final feed). In this experiment, the Vp28 fusion is a recombinant polypeptide prepared by Invitrogen's PurePro Caulobacter Expression System, in which Vp28 fragment 1E (SEQ ID NO: 4) is fused to the surface array protein RsaA from Caulobacter crese...
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