• International Journal of Industrial Entomology and Biomaterials
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• v.14 no.1
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• pp.51-56
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• 2007

In a previous study, three larval cuticle protein genes were cloned from the mulberry longicorn beetle, Apriona germari (Comp. Biochem. Physiol. B 136, 803-811, 2003). In the present study, the genomic structures of these three larval cuticle protein genes (AgLCP9.2, AgLCP12.6 and AgLCP12.3) were elucidated. All three cuticle protein genes consist of one intron and two exons. Southern blot analysis of genomic DNA suggested that three cuticle protein genes are a single copy gene. In addition, a novel larval cuticle protein gene, AgLCP10.6, was cloned from A. germari in this study. The AgLCP10.6 cDNA contains an ORF of 300 nucleotides that are capable of encoding a 100-amino acid polypeptide with a predicted molecular mass of 10.6 kDa. The amino acid sequence deduced from the AgLCP10.6 cDNA contained a type-specific consensus sequence identifiable in other insect cuticle proteins and is most homologous to Drosophila melanogaster cuticle protein ACP65A (51 % protein sequence identity). Northern blot analysis revealed that AgLCP10.6 showed epidermis-specific expression.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.223-229
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• 2007

Present study aims to investigate the topical distribution of pupal stage specific cuticle protein and its temporal and spatial role during the wing formation of Artogeia rapae. ArCP27(27 kd cuticle protein) was identified as pupal stage specific cuticle protein in cuticle tissues and has not shown any qualitative differences by local portions of body. ArCP27 maintained constant concentration just after pupal ecdysis to 5-day old pupal stage but thereafter decreased. In fat body, ArCP27 was found in both thoracic and abdominal fat body from the last larval to pupal stage. In wing cuticle, ArCP27 began to find from 5-day old pupal stage. Immunologically ArCP27 in thoracic and abdominal cuticle has the response against the ArCP27 at 5-day old pupa but since then has no response. But the antibody against ArCP27 has reacted to 5- and 7-day old pupal and adult wing protein. $^3H-leucine$ was not incorporated into ArCP27 in 5- and 7-day old thoracic and abdominal cuticle but was incorporated into ArCP27 in 7-day old wing cuticle and adult wing, suggesting that ArCP27 partly participates the wing cuticle formation by the process of digestion and reabsorption of old cuticle.

• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.1
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• pp.67-70
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• 2005

A cuticle protein gene, PbLCP12.1, from the white­spotted flower chafer, Protaetia brevitarsis, was isolated and characterized. The gene contains an ORF of 336 nucleotides capable of encoding a 113 amino acid polypeptide with a predicted molecular mass of 12,138 Da and pI of 4.15. The PbLCP12.1 protein contained a type-specific consensus sequence identifiable in other insect cuticle proteins. The deduced amino acid sequence of the PbLCP12.1 cDNA is most similar to Bombyx mori cuticle protein BmLCP18 (37$\%$ protein sequence identity). Northern blot analysis revealed that PbLCP12.1 showed the epidermis-specific expression.

• Applied Microscopy
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• v.37 no.1
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• pp.43-52
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• 2007

In order to observe the localization of excretory, purified and infected antigenic protein in the tissue of Trichinella spiralis larvae, immunogoldlabeling methodology using IgG and protein A-gold complex was implemented. T. spiralis larvae obtained from rat muscle were initially cultured in medium, and secreted excretory antigen was collected for 1 or 3 days. Purified antigenic protein was obtained from homogenized T. spiralis larvae. Rabbits were then immunized with 1 or 3 days secreted excretory protein and purified 45 kDa protein, and IgG was purified from collected serum. Serum, against infected antigen, collected from rat on 1 and 4 weeks after infection with T. spiralis larvae, and IgG was purified from collected serum. T. spiralis larvae were embedded in Lowicryl HM20 medium. Then they were finally treated with immunized IgG and protein A-gold complex (particle size; 15 nm) and observed under electron microscope. In T. spiralis larvae tissue, the tissue antigen reacted with rabbit IgC antigen Day 1 secreted excretory protein, infected antigenic protein and purified 45 kDa protein. But different distribution pattern of labeled gold particles were observed. When Day 1 secreted excretoy protein was used, gold particle labeling was observed specifically on the cuticle, basal layer, esophagus interstitial matrix (EIM) and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. In a separate group of tissue, the antigen reacted with rabbit IgG against Day 3 secreted excretory protein. Labeled gold particles were specifically distributed on the surface layer of cuticle, EIM and ${\alpha}_0$ granules of stichocyte of the worm. In case of using infected antigenic protein, gold particle labeling was specifically distributed on the cuticle and EIM of the worm. When purifed 45 kDa protein was used gold particle labeling was specifically distributed on the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. Therefore, excretory antigens appeared to originate from the cuticle and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte for the first day but the cuticle layer associated with globular proteins and ${\alpha}_0$ granules of stichocyte after 3 days and infected antigens appeared to originate from the cuticle for 1 and 4 weeks after infection. These results suggest that excretory and infection specific antigens are secreted into the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte and 45 kDa protein may be contained these specific antigens.

• International Journal of Industrial Entomology and Biomaterials
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• v.10 no.1
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• pp.11-17
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• 2005

In our research to identify gene involved in the cuticle protein, we cloned a novel cuticle protein gene, ApCP15.5, from the Chinese oak silkmoth, Antheraea pernyi, larvae cDNA library. The gene encodes a 149 amino acid polypeptide with a predicted molecular mass of 15.5 kDa and a pI of 9.54. The ApCP15.5 contained a type-specific consensus sequence identifiable in other insect cuticle proteins and the deduced amino acid sequence of the ApCP15.5 cDNA is most homologous to Tenebrio molitor-C1B ($43\%$ protein sequence identity), followed by Locusta migratoria-76 ($42\%$ protein sequence identity). Northern blot and Western blot analyses revealed that the ApCP15.5 showed the epidermis-specific expression. The expression profile of ApCP15.5 indicated that the ApCP15.5 mRNA expression was detected in the early stages after larval ecdysis and larval-pupal metamorphosis, and its expression level was most significant on the first day of larval ecdysis and pupal stage. The ApCP15.5 was expressed as a 15.5 kDa polypeptide in baculovirus-infected insect cells.

• The Korean Journal of Zoology
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• v.23 no.1
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• pp.1-12
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• 1980

Changes and possible origin of haemolymph proteins during the cuticle formation and hardening are determined by means of acrylamide gel electrophoresis and immunodiffusion. The results by acrylamide gel electrophoresis showed at least 19 protein bands in the haemolymph and 13 fractions in the fat body with relatively constant pattern during the period of cuticle formation and hardening. Both haemolymph and fat body proteins are generally characterized by the presence of three to four heavy stained bands and several thin bands near the top region of the gel. At least over five haemolymph proteins are constantly present during this period. Immunodiffusion tests show that of total eight to nine pupal haemolymph proteins two proteins were already detected in the fat body before pupation and other two proteins were also found in the fat body immediately after pupation, suggesting fat body as possible source of these two haemolymph proteins.

• Journal of the Korean Society of Clothing and Textiles
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• v.46 no.3
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• pp.494-512
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• 2022

Hair toners containing polyphenol or Vitamin B5 were investigated according to their recovering effects on hair damaged by bleaching. Surface morphology, CIE L*a*b* values, and tensile properties of hair were measured. The amount of protein leaking from hair was investigated using the Bradford protein assay. The amino acid composition of hair was examined using the HPLC instrument. Hair became severely damaged after bleaching, showing cuticle structure with surface melt down and rolled up tip, a decrease in tensile strength, an increase in protein leak, and an increase in the proportion of cysteic acid. When bleached hair was treated with the two types of hair toner, positive effects were seen in the recovery of cuticle structure and retention of bleached color, an increase in tensile strength, a decrease in protein leak up to certain days, and an increase in the retention of protein examined by the HPLC analysis of amino acids. Hair treated with B5 toner showed better effects on the increase of tensile strength compared to the hair treated with PP toner. Hair treated with PP toner showed better retention of color, less protein leak, and a lower proportion of cysteic acid compared to the hair treated with B5 toner.

• The Korean Journal of Zoology
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• v.22 no.1
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• pp.19-25
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• 1979

To determine the metabolism of leucine during the cuticle formation and the sclerotization process in Pieris rapae L., $U-^3$H-leucine or $U-^4$C-tyrosine is injected into the haemolymph of newly molted pupa through dorsal cuticle of heart area. The results show that leucine as a common amino acid participates in the synthesis of cuticle protein over the first 3 hr after ecdysis. It is also shown that leucine in the haemolymph at ecdysis is freely being moved between major internal organs during the short time period post-ecdysis, providing the evidence for some involvements including haemolymph protein synthesis and storage of fat body and gut in metabolism of leucine.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2003.10a
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• pp.134-135
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• 2003

The insect cuticle undergoes drastic morphological alterations during postembryonic development and is an extracellular structure composed mainly of chitin and proteins that are synthesized and secreted by epidermal cells. Cuticle proteins, the major components of insect integument, are recently being focused as a model fur studying the mechanisms of gene regulation during molting and metamorphosis. (omitted)

• Journal of Plant Biology
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• v.39 no.3
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• pp.189-195
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• 1996

The major cuticular components have been shown to be synthesized in the epidermis. Therefore, cloning of epidermis-specific genes could yield information to be used to isolate and characterize the enzymes involved in the cuticle biosynthesis. A subtractive cDNA library was prepared from Senecio odorus in which epidermis-specific cDNAs were enriched. Differential screening of the library using epidermal and non-epidermal probes revealed two cDNAs. One of them designated epi425 was identified, based on the sequence homology, as a member of a new class in the LTP gene family and the other clone designated epi23 as a gene encoding an aldehyde decarbonylase. Northern blot analyses showed that epi425 and epi23 cDNAs hybridized with a transcript of about 600 and 2, 100 nucleotides, respectively, from the epidermis but not from the non-epidermal tissues. Further characterization of these clones will provide more information on the mechanism of the cuticle biosynthesis.