• Title/Summary/Keyword: Silkworm (Bombyx mori)

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Characterization of Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Bombyx mori Nucleopolyhedrovirus

  • Zhang, Zhi-Fang;Shen, Xing-Jia;Yi, Yong-Zhu;Tang, Shun-Ming;Li, Yi-Ren;He, Jia-Lu;Wu, Xiang-Fu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.169-174
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    • 2004
  • Bombyx mori nucleopolyhedrovirus(BmNPV) ecdysteroid UDP-glucosyltransferase gene (egt) promoter fragments of different lengths were amplified from BmNPV ZJ-8 genomic DNA by PCR. Reporter plasmids pBmegt542-luc, pBmegt309-luc and pBmegtl59-luc with luciferase (lue) driven by egt promoters were constructed. Both in vitro and in vivo expressions showed that BmNPV egt promoter activity requires the transactivation of viral factor(s), and expression of luc was detected earliest at 24 hrs post infection (pi). BmNPV ZJ-8 homologous region 3 (hr3) increased the expression of luc by over 1,600-fold. Molting hormone of 1.0 - 2.0 $\mu\textrm{g}$/$m\ell$ can dramatically down regulate expression of luc. Juvenile hormone analogue of 0.5-2.0 ${\mu}g$/$m\ell$ increased expression of luc by 145.8% to 75.7%. Deletion assay revealed that the promoter fragment of 159 bp contains the basal promoter structure; Promoter fragments of 309 bp and 542 bp showed similar but much higher transcriptional activities than that of 159 bp, suggesting that nucleotide from -159 to -309 nt upstream the translation initiation site harbors the main cis-acting elements.

Identification of Dopa decarboxylase associated protein from Bombyx mori

  • Hwang, Jae-Sam;Kim, Sung-Kuk;Kang, Seok-Woo;Goo, Tae-Won;Yun, Eun-Young;Park, Kwang-Ho;Chang, Jong-Soo
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.97-98
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    • 2003
  • Current study was aimed to understand an interaction between Dopa decarboxylase (DDC) and proteins that specifically binds to DDC in the silkworm, Bombyx mori. Materials and Methods: Materials-Animal: Bombyx mori Construction of GST fusion protein Preparation of lysates: Protein extracted from whole body of Bombyx mori Methods-In vitro binding assay with lysates, Peptide sequence and RACE-PCR (omitted)

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Cloning and Characterization of hydroxypyruvate isomerase (EC 5.3.1.22) gene in silkworm Bombyx mori

  • Lv, HongGang;Chen, KePing;Yao, Qin;Wang, Lin
    • International Journal of Industrial Entomology and Biomaterials
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    • v.17 no.2
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    • pp.189-195
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    • 2008
  • The sequence of hydroxypyruvate isomerase gene was obtained in NCBI. In this study, the hydroxypyruvate isomerase gene of Bombyx.mori was identified and annotated with bioinformatics tools. The result was confirmed by RT-PCR, prokaryotic expression, mass spectrographic analysis and sub-cellular localization. The hydroxypyruvate isomerase cDNA comtains a 783bp ORF, and has 4 exons. The deduced protein has 260 amino acid residues with the predicted molecular weight of 29169.30 Da, isoelectric point of 6.10, and contains conserved PRK09997 and Hfi domains. The hydroxypyruvate isomerases of Nasonia vitripennis and Bombyx mori have a high homology. Through RTPCR analysis, we found that this transcript was present in testis, ovary, blood-lymph, fat body, midgut, silk gland and tuba Malpighii. This protein was located in cytoplasm through immunohistochemistry. We submitted the cloned gene under the accession number EU344910. The enzyme has been classified under accession number EC 5.3.1.22.

Characteristics and pathogenicity of the Entomopathogenic Fungus Beauveria bassiana101AA on the silkworm (Bombyx mori) (백강균(Beauveria bassiana)101A의 특성 및 가잠(Bombyx mori)에 대한 병원성 검정)

  • 정이연;남성희;조세연
    • Journal of Sericultural and Entomological Science
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    • v.42 no.2
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    • pp.99-103
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    • 2000
  • This study was carried out to investigate incubating characteristics, LTSEM observation, nature of a disease of infected silkworms(bombyx mori), LD$\_$50/, optimum infective condition for mass production and infection percentage(%) on conservation periods. On the V8 media, the colony was 0.5∼2.0cm/8 days in diameter and white or slighty colored with a white fluffy to powdery appearance. The conidia was subglobos in shape, zig-zag appearance, 1-celled, hyaline and 3.2$\times$2.4 ㎛ in size on the average. The conidiophore was irregular grouped, hyaline, rounded or flask-shaped. The LD$\_$50/ values of the 2nd and 4th silkworms were each other 6.6142(Log) in natural temperature and humidity (25$\^{C}$, 65%). On optimum infective condition for mass production, all of preservation time(hr.) postinoculation in 1.0$\times$10$\^$8/ conidia/㎖ was over 97% and only 20 hr in 1.0$\times$10$\^$7/ condia/㎖ was over 90%.

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Cloning, Sequencing and Characterization of Mitochondrial Control Region of the Domestic Silkwom, Bombyx mori

  • Lee, Jin-Sung;Kim, Ki-Hwan;Hoe, Hyang-Sook;Park, Jae-Heung;Kang, Seok-Woo;Lee, Sang-Han;Hwang, Jae-Sam
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.1
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    • pp.87-89
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    • 2001
  • The nucleotide sequence of the domestic silkworm (Bombyx mori) mitochondrial (mt) control region and its flanking genes was determined from PCR clones. The control region of the silkworm mt genome was located between the small ribosomal RNA gene and transfer RN $A^{Met}$. This 499 bp control region hale 95.4% A+T content. Extensive comparative analysis studies performed with similar control region of other insect genomes could not reveal a highly conserved region containing conserved motifs of animal mito-chondrial genome. The remarkable feature that found in this control region was the presence of tandem motifs containing nine repetitive sequences. The potential usefulness of this motif sequences for Bombyx species or their taxonomically related species is enhanced by its unique localization in the maternally inheritance mitochondrial molecule.e.

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Evaluation of Different Bed Disinfectants against the Spread of Common Diseases in Silkworm, Bombyx mori L.

  • Irfan Illahi;S. D. Sharma;K. Chrasekharan;B. Nataraju;M. Balavenkatasubbaiah;T. Selvakumar;V. Thiagarajan;S. B. Dandin
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.2
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    • pp.191-196
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    • 2003
  • Silkworm body and rearing seat disinfection is an integral part of effective silkworm rearing as it prevents the spread of various diseases through secondary contamination in the rearing bed. Many bed disinfectants are recommended by various research institutes in India. In the present study, eight bed disinfectants viz., Vijetha, Labex, Resham Jyothi, Sanjeevini, Suraksha, Reshamkeet Oushadh, Dithane M45 and Captan recommended by various Sericultural Research Institutions in the country against the spread of common diseases of silkworm were tested to determine their comparative efficacy. For the study, all the treatment batches were introduced with fixed number of specific diseased larvae so as to maintain a constant source of pathogen in the bed and then followed by treatment with respective bed disinfectants. Percent mortality/infection due to each disease was recorded to determine efficacy of bed disinfectant treatment against respective disease. The results show that all the bed disinfectants tested have considerable efficacy against the spread of various diseases in the rearing bed. However, Vijetha was distinctly superior in its efficacy against all four major diseases of silkworm. The study emphasize the role of bed disinfectants in silkworm rearing and the fact that they should be judiciously selected based on the efficacy of the bed disinfectant and the type of disease prevalent in a particular season and area.

Colloidal Textile Dye-Based Dipstick Immunoassay for the Detection of Infectious Flacherie of Silkworm, Bombyx mori L.

  • Sivaprasad, V.;Nataraju, B.;Renu, S.;Datta, R.K.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.6 no.1
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    • pp.27-31
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    • 2003
  • Infectious flacherie of silkworm Bombyx mori is caused by B. mori infectious flacherie virus (BmIFV) and causes severe crop loss to sericulturists. In the present study, a colloidal textile dye-based dipstick immunoassay is developed for the detection of infectious flacherie in silkworms. Colloidal textile dye (blue D2R) with Aλ$_{max}$ at 620 nm was sensitised with 500 $\mu\textrm{g}$/ml of purified anti-BmIFV IgG. The dye-antibody reagent detects purified antigen up to 10 ng/ml and BmIFV infection in diseased larval extracts $(up to a dilution of {10^-5})$ and faecal matter extracts $(up to a dilution of {10^-2})$ by forming clear blue dot within 30 min. It was observed to be stable for three months period at $4^{\circ}C$. The efficacy of textile dye-based dipstick immunoassay was on pay with HRP-based dipstick immunoassay and fluorescent antibody test, and better than latex agglutination and ouchterlony tests in the detection of BmIFV The dye-based dipstick immunoassay method provides a simple, sensitive and less expensive test for the detection of BmIFV infection in silkworms.s.

Seasonal Impact of Microsporidian Infection on the Reproductive Potential of Silkworm, Bombyx mori L. (Lepidoptera: Bombycidae)

  • Mohanan N. Madana;Krishnan N.;Mitra P.;Das N. K.;Saratchandra B.;Haldar D. P.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.2
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    • pp.107-111
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    • 2005
  • Impact of microsporidian infection and season on reproductive potential of Bombyx mori L. was investigated in the laboratory. Microsporidian infection significantly (P<0.01) reduced fecundity and hatching and increased sterility and mortality of eggs. Among the microsporidia, Nosema sp. 2 infected silkworm produced eggs with least fecundity and hatching percent as well as highest dead and sterile eggs followed by Nosema sp. 1 and N. bombycis. Microsporidia, in general, significantly reduced fecundity and hatching percent of eggs and increased number of dead and sterile eggs in all the three seasons except N. bombycis in July - August (S3) and Nosema sp. 2 in January February (S1). Since, seed production is the anchor sheet of mulberry sericulture, coefficient of egg lying is considered as an important aspect and the industry quite often facing shortage of disease free layings. The present study indicates that B. mori is more susceptible to microsporidia during S2 followed by S3 and S1 and Nosema sp2. is most virulent followed by Nosema spl. and N. bombycis.

Insect Adaptations to Changing Environments - Temperature and Humidity

  • Singh, Tribhuwan;Bhat, Madan Mohan;Khan, Mohammad Ashraf
    • International Journal of Industrial Entomology and Biomaterials
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    • v.19 no.1
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    • pp.155-164
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    • 2009
  • The most important factors in environment that influence the physiology of insects are temperature and humidity. Insects display a remarkable range of adaptations to changing environments and maintain their internal temperature (thermoregulation) and water content within tolerable limits, despite wide fluctuations in their surroundings. Adaptation is a complex and dynamic state that widely differs in species. Surviving under changing environment in insects depends on dispersal, habitat selection, habitat modification, relationship with ice and water, resistance to cold, diapause and developmental rate, sensitivity to environmental signals and syntheses of variety of cryoprotectant molecules. The mulberry silkworm (Bombyx mori) is very delicate and sensitive to environmental fluctuations and unable to survive naturally because of their domestication since ancient times. Thus, the adaptability to environmental conditions in the silkworm is quite different from those of wild insects. Temperature, humidity, air circulation, gases and photoperiod etc. shows a significant interaction in their effect on the physiology of silkworm depending upon the combination of factors and developmental stage affecting growth, development, productivity and quality of silk. An attempt has been made in this article to briefly discuss adaptation in insects with special emphasis on the role of environmental factors and their fluctuations and its significance in the physiology of mulberry silkworm, B. mori.

Hemocyte-specific Promoter for the Development of Transgenic Silkworm, Bombyx mori

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong-Ryul;Choi, Gwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.1
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    • pp.111-114
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    • 2012
  • In previous studies we have shown that a sw17255 gene was expressed in hemocyte-specific tissues of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). It was verified that the sw17255 core promoter region contains elements that regulate the expression of this gene in hemocyte tissue; the selected promoter region spans nucleotides -1 to -2,112 upstream of the start codon. Each of the luciferase reporter gene expression vectors under the control of 4 different kinds of promoter candidates, (-2,112/-1), (-1,640/-1), (-1,169/-1) and (-579/-1), and the control reporter plasmid DNA, were introduced into B. mori larval coelom by direct injection using a syringe. The promoter candidate [E] (-579/-1) showed more than 1.67 fold transcriptional activity compared to control promoter activity. Higher productivity of an expressed gene in the transgenic silkworm by this promoter combination could be achieved in the near future. The foreign recombinant protein could be easily harvested from the blood of the transgenic silkworm.