Gene Expression of a Novel Defensin Antimicrobial Peptide in the Silkworm,<i>Bombyx mori</i>

Kaneko, Yasunari; Tanaka, Hiromitsu; Ishibashi, Jun Ichiro; Iwasaki, Takashi; Yamakawa, Minoru

doi:10.1271/bbb.80263

Cited by 40 publications

(27 citation statements)

References 45 publications

(47 reference statements)

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“…1) provided that it form a cluster distinct from the previously described insect defensins. It showed association with defensinlike molecules especially within the Lepidoptera order as previous study (Volkoff et al, 2003), and suggested that BmDefensinA and BmDefensinB gene evolved from different ancestral genes, supporting our observation by other studies of BmdefA and BmdefB in B. mori (Kaneko et al, 2008;Wen et al, 2009).…”

Section: Discussionsupporting

confidence: 91%

“…This finding would seem that Bombyx defensin-like protein is functionally responsible for antibacterial activity. BmDefA, however, was only slightly activated by E. coli, B. subtilis, and B. bassiana, whereas BmDefB was strongly activated by these microorganisms (Kaneko et al, 2008). Thus it indicated that BmDefensinB rather than BmDefensinA plays a major role in defense against microbial infection.…”

Section: Discussionmentioning

confidence: 91%

“…In a recent study, the amino acid sequence of BmDefensin B showed relatively low similarity(about 27%) with BmDefensin A, which is designated as BmDp in this study. Phlylogenetic analysis suggested that BmDefensinB (B. mori defensin like protein 2) and BmDefensinA evolved independently from different ancestral genes (Kaneko et al, 2008). According to phylogenetic tree analysis (Fig.…”

Section: Amino Acid Sequence and Phylogeny Analysismentioning

confidence: 99%

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Molecular characterization of anionic defensin-like peptide in immune response of silkworm, Bombyx mori L. (Lepidoptera)

et al. 2010

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We have previously identified a defensin-like peptide (BmDp, Genbank accession number DQ118523), in the larvae of a silkworm, Bombyx mori (B. mori) using wdS20658 cDNA in a Bombyx EST database (Silk Base: http://www.ab.a.u-tokyo.ac.jp/silkbase). A full-length of BmDp cDNA was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) and expressed in Escherichia coli cells in order to obtain a functional peptide. Amino acid sequence comparison of cysteine-rich defensin-like peptides with those of the BmDp showed to be identical protein as BmDefensinA designated as BmdefA gene deposited in the DDBJ database under the accession number AB367525. Sequence alignments revealed that the BmDp belongs to the insect defensin family and is close to galiomicin and spodoptericin, a defensin and a defensin-like protein having antimicrobial activities in phylogenetic analysis. Thus, we evaluated the effect of immune challenge and antimicrobial activity to understand the nature of this anionic BmDp. Semiquantitative RT-PCR analysis showed that BmDp gene expression was inducible by bacterial injection and peaked at a highest level at 8h after bacterial injection. This result suggests that BmDp is related with immune response against bacteria. A purified recombinant GSTBmDp fusion protein did not show direct antimicrobial activity against bacteria and fungi, which is differed from cationic antibacterial insect defensins and neutral antifungal defensin-like peptides.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 91%

Section: Amino Acid Sequence and Phylogeny Analysismentioning

confidence: 99%

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Molecular characterization of anionic defensin-like peptide in immune response of silkworm, Bombyx mori L. (Lepidoptera)

et al. 2010

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“…Produced by a wide variety of organisms as part of a non-specific immune response, these peptides are involved in the directed destruction of various microorganisms. In B. mori, six different families of antimicrobial peptides have been identified: cecropin, attacin, lebocin, moricin, goverin, and defensin (Steiner et al, 1981;Sugiyama et al, 1995;Tanaka et al, 2008;Hara et al, 1995;Kaneko et al, 2007;Kaneko et al, 2008). The induction mechanism of these genes is divided into two distinct pathways: the Toll and IMD pathways (Lemaitre et al, 2007).…”

Section: Discussionmentioning

confidence: 99%

Production of the BmCecB1 antimicrobial peptide in transgenic silkworm

Kim¹,

Kim²,

Park³

et al. 2015

International Journal of Industrial Entomology

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This peptide has antibacterial activity against several Gram-positive and Gram-negative bacteria. Bombyx mori cecropinB1(BmCecB1) is antimicrobial peptides from Bombyx mori and belongs to cecropin family. Antimicrobial peptides are important components of the innate immune systems in all living organism. To produce the BmCecB1 antimicrobial peptide, we constructed transgenic silkworm that expressed BmCecB1 gene under the control BmA3 promoter using piggyBac vector. The use of the 3xP3-driven EGFP cDNA as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor vector and helper vector were micro-injected into 600 eggs of bivoltin silkworms, Baegokjam. In total, 49 larvae (G0) were hatched and allowed to develop into moths. The resulting G1 generation consisted of 22 broods, and we selected 2 broods containing at least 1 EGFP-positive embryo. The rate of successful transgenesis for the G1 broods was 9%. We identified 9 EGFP-positive G1 moths and these were backcrossed with wild-type moths. With the aim of identifying a BmCecB1 as antimicrobial peptide, we investigated the Radical diffusion Assay (RDA) and then demonstrated that BmCecB1 possesses high antibacterial activities against Gramnegative bacteria. Recently, a germline transformation method using the GCTAGCATGCGTGTTTTAATTTTCACG-3' and reverse primer 5'-CTTGGTTCGGCTAAAGCTATACTTAAG-3') into

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“…The molecular mechanisms of the insect's innate immune system have been studied using both molecular and proteomic methods (7,8). Drosophila melanogaster is a model organism, and there is a vast amount of information available on its genetics as well as the molecular mechanisms of its immune response (9).…”

Section: Introductionmentioning

confidence: 99%

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Silva

Barbosa

Bravo

et al. 2010

Braz J Med Biol Res

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Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

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Gene Expression of a Novel Defensin Antimicrobial Peptide in the Silkworm,Bombyx mori

Cited by 40 publications

References 45 publications

Molecular characterization of anionic defensin-like peptide in immune response of silkworm, Bombyx mori L. (Lepidoptera)

Molecular characterization of anionic defensin-like peptide in immune response of silkworm, Bombyx mori L. (Lepidoptera)

Production of the BmCecB1 antimicrobial peptide in transgenic silkworm

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

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