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[en] Males and females of a Bt-tolerant mulberry silkworm (Bombyx mori L.) population were crossed with females and males of a Bt-susceptible population, to produce Bt-tolerant silkworm hybrids, and to determine the expression of the Bt-tolerance pattern in the F 1 hybrids. It was observed that when a Bt-tolerant (42% larval mortality) female (BtT ) silkworm was crossed with a Bt-susceptible (85% larval mortality) male (BtS ), the resultant F 1 offspring showed lower levels of Bt-tolerance (87% larval mortality). On the other hand, when a Bt-tolerant male (BtT ) was crossed with a Bt-susceptible female (BtS ), the F 1 hybrid showed higher levels of Bt-tolerance (35% larval mortality) characteristic. The probit statistics showed that both hybrids expressed Bt-tolerance or susceptible levels similar to their male parents. These different patterns of Bt-tolerance in F 1 hybrids might be due to the transferring of a Bt-tolerant gene, from the parents to offspring, through the homozygotic male (ZZ) silkworm. (author)
[en] We constructed the fibroin H-chain expression system to produce recombinant proteins in the cocoon of transgenic silkworms. Feline interferon (FeIFN) was used for production and to assess the quality of the product. Two types of FeIFN fusion protein, each with N- and C-terminal sequences of the fibroin H-chain, were designed to be secreted into the lumen of the posterior silk glands. The expression of the FeIFN/H-chain fusion gene was regulated by the fibroin H-chain promoter domain. The transgenic silkworms introduced these constructs with the piggyBac transposon-derived vector, which produced the normal sized cocoons containing each FeIFN/H-chain fusion protein. Although the native-protein produced by transgenic silkworms have almost no antiviral activity, the proteins after the treatment with PreScission protease to eliminate fibroin H-chain derived N- and C-terminal sequences from the products, had very high antiviral activity. This H-chain expression system, using transgenic silkworms, could be an alternative method to produce an active recombinant protein and silk-based biomaterials
[en] We have previously shown that budded viruses of Bombyx mori nucleopolyhedrovirus (BmNPV) enter the cell cytoplasm but do not migrate into the nuclei of non-permissive Sf9 cells that support a high titer of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) multiplication. Here we show, using the syncytium formation assay, that low-pH-triggered membrane fusion of BmNPV GP64 protein (Bm-GP64) is significantly lower than that of AcMNPV GP64 protein (Ac-GP64). Mutational analyses of GP64 proteins revealed that a single amino acid substitution between Ac-GP64 H155 and Bm-GP64 Y153 can have significant positive or negative effects on membrane fusion activity. Studies using bacmid-based GP64 recombinant AcMNPV harboring point-mutated ac-gp64 and bm-gp64 genes showed that Ac-GP64 H155Y and Bm-GP64 Y153H substitutions decreased and increased, respectively, the multiplication and cell-to-cell spread of progeny viruses. These results indicate that Ac-GP64 H155 facilitates the low-pH-triggered membrane fusion reaction between virus envelopes and endosomal membranes.
[en] Experiments were conducted to study the effect of gamma radiation on larvae of silkworm for increased production of silk. Irradiated larvae showed an increase in cocoon shell weight compared to unirradiated ones. The highest percentage (17.50%) of silk increased was recorded at 5 Gy in fourth instar larvae and the lowest was 0.62% at 6 Gy in the third instar larvae of silkworm. The average highest shell weight (0.141 g) and filament length (527.09 metre) were also observed at 5 Gy in fourth instar larvae. The lowest shell weight and filament length were noted in untreated batch of the respective instars of larvae.(author)
[en] A total of 24 isolates of Bacillus thuringiensis (Berliner), resistant to a γ-radiation dose of 100 krad, were screened for their toxicity to larval silkworms, Bombyxmori(L.), and 15 of them were subsequently tested for their toxicity to larval diamond-back moth, Plutella xylostella(L.). The LC50's of these isolates to B. mori ranged from 1.6 X 105 to 6.0 X 103 spores/mL or from 5.9 to 0.3 μg cellular protein/mL. The irradiation treatment produced isolates which were significantly more toxic to P. xylostella (LC50 < 8.1 X 104 spores/mL or 3.7 μg cellular protein/mL) and/ or less toxic to B. mori (LC50 > 2.3 X 104 spores/mL or 1.0 μg cellular protein/mL) than the parent commercial strain
[en] Silkworms, Bombyx mori, are a promising model animal in health safety and environmental pollution assessment due to their sensitivity to chemical compounds like pesticides, drugs, and heavy metals, in addition to other features like their low cost and body characteristics and their full genome sequencing. In this review, we summarize the silkworm advantages as a model organism in toxicological research. .
[en] A protein showing strong antiviral activity against Bombyx mori nucleopolyhedrovirus (BmNPV) was purified from the digestive juice of B. mori larvae. The molecular mass of this protein was 24 271 Da. Partial N-terminal amino acid sequence of the protein was determined and cDNA was cloned based on the amino acid sequence. A homology search of the deduced amino acid sequence of the cDNA showed 94% identity with B. mori serine protease so the protein was designated B. mori serine protease-2 (BmSP-2). Analysis of BmSP-2 gene expression showed that this gene is expressed in the midgut but not in other tissues. In addition, BmSP-2 gene was shown to not be expressed in the molting and wandering stages, indicating that the gene is hormonally regulated. Our results suggest that BmSP-2, an insect digestive enzyme, can be a potential antiviral factor against BmNPV at the initial site of viral infection
[en] The tests involving the hybrids 48-I x 157-K and J-124 x C-122 were conducted at the experimental base of the Sericulture Experiment Station in Vratza. Silkworm seed was irradiated once daily from the first day of incubation to whitening of the seed at 60 and 80 min exposure. The combined quartz lamp, type ''MLKK-5'' (500 W, 220 V) was used. Seed was irradiated at a distance of 100 cm from the burner after its preliminary heating in the course of at least 10 min. Silkworm infrared irradiation exerts a stimulatory effect on embryonal and larval development, silkworm viability, and row cocoon yield. Sixty minutes irradiation exposure proved to be more effective for the native hybrid 48-I x 157-K, silkworm viability increasing by 3.75% and row cocoon yield per g silkworm seed - by 0.37 kg. On the contrary, 80 min irradiation exposure proved more effective in the case of the Japanese hybrid J-124 x C-122. (B.Ch.)
[en] Despite close genetic relationship, Bombyx mori nucleopolyhedrovirus (BmNPV) and Autographa californica multicapsid NPV (AcMNPV) display a distinct host range property. Here, BmNPV replication was examined in Sf9 and High Five cells that were nonproductive for BmNPV infection but supported high titers of AcMNPV replication. Recombinant BmNPV, vBm/gfp/lac, containing bm-ie1 promoter-driven egfp showed that few Sf9 and High Five cells infected with vBm/gfp/lac expressed EGFP, while large proportion of EGFP-expressing cells was observed when transfected with vBm/gfp/lac DNA. Immunocytochemical analysis showed that BmNPV was not imported into the nucleus of these two cell lines, while recombinant BmNPV, vBmΔ64/ac-gp64 possessing AcMNPV gp64 was imported into the nucleus, yielding progeny virions in High Five cells, but not Sf9 cells. These results indicate that the defective nuclear import of infected virions due to insufficient BmNPV GP64 function is involved in the restricted BmNPV replication in Sf9 and High Five cells