[논문]BmNPV Infection Enhances Ubiquitin-conjugating Enzyme E2 Expression in the Midgut of BmNPV Susceptible Silkworm Strain

Gao, Lu; Chen, Keping; Yao, Qin; Chen, Huiqing

BmNPV Infection Enhances Ubiquitin-conjugating Enzyme E2 Expression in the Midgut of BmNPV Susceptible Silkworm Strain 원문보기

International journal of industrial entomology, v.13 no.1, 2006년, pp.31 - 35

Gao, Lu (School of Medical Technology, Jiangsu University) , Chen, Keping (Institute of Life Sciences, JiangSu University) , Yao, Qin (Institute of Life Sciences, JiangSu University) , Chen, Huiqing (Institute of Life Sciences, JiangSu University)

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The ubiquitin conjugating enzyme 2 (E2) is core component of ubiquitin proteasome pathway (UPP) which represents a selective mechanism for intracellular proteolysis in eukaryotic cells. The E2 has been implicated in the intracellular transfer of ubiquitin to target protein. We show here the involvement of E2 in antiviral immune of Bombyx mori to Bombyx mori nuclear polyhedrosis virus (BmNPV). In this study, mRNA fluorescent differential display PCR (FDD-PCR) was performed with BmNPV highly resistant silkworm strain NB and susceptible silkworm strain 306. At 24 h post BmNPV infection, FDD-PCR with the arbitrary primer AP34 showed that one cDNA band was down-regulated in the midgut of resistant strain, but highly expressed in susceptible strain. The deduced amino acid sequence of this cDNA clone share 99% identity with the recently published B. mori ubiquitin conjugating enzyme E2 (Genbank NO: DQ311351). Fluorescent quantitative PCR corroborated down regulation of E2 in resistant strain. We there conclude that BmNPV infection evokes strong response of susceptible strain including activation of UPP. BmNPV may evolve escape mechanisms that manipulate the UPP in order to persist in the infected host. In addition, the identification of down-regulation of E2 in resistant strain, as well as structure data, are essential to understanding how UPP operates in silkworm antiviral immune to BmNPV disease.

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The primers were designed according to the sequences of Bombyx mori Ubc E2 (Genbank NO: DQ311351). The sense primer for E2 was 55 -TACAGAACGGCGACAT-TGATCT-3' and the antisense primer, 5'・ATTAGCCTG-TAGCCACCTCACC-3, .

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where R is the relative expression ratio. Statistical analysis of differences between the groups was carried out using the t-test. P values of <0.

이론/모형

To comfirm that the results from differential display, we carried out fluorescent Real-time PCR assay. The more relible real-time PCR results showed that the level of mRNA expression for E2 of strain 306 increased by _사times over strain NB upon BmNPV challenge (P<0.
Translation into amino-acid sequence was performed with ExPASy Translate tool (http://www.expasy.ch/tools/dna. html). The multisequence alignment was performed by the CLUSTAL W program, using the following sequences: D.

성능/효과

1). Databases searching showed that this cDNA exhibited a sequence similarity to B. mori EST clone CK53_사225 and CK487465. By assembling these nucleotide sequences, we fb_니nd that it was 936 bp in length plus a polyadenylation.
out fluorescent Real-time PCR assay. The more relible real-time PCR results showed that the level of mRNA expression for E2 of strain 306 increased by _사times over strain NB upon BmNPV challenge (P<0.05), similar to the FDD-PCR result (Fig. 2). It appears that the E2 gene of highly susceptible silkworm 306 responded more strongly to the virus invasion than that of strain NB at the early phase of infection.
To confirm further that the newly identified EST clone belongs to the silkworm homolog of human E2, the deduced amino acid sequence of this clone was used tocompare E2 derived from various species. As shown in Fig.

참고문헌 (17)

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BmNPV Infection Enhances Ubiquitin-conjugating Enzyme E2 Expression in the Midgut of BmNPV Susceptible Silkworm Strain 원문보기

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BmNPV Infection Enhances Ubiquitin-conjugating Enzyme E2 Expression in the Midgut of BmNPV Susceptible Silkworm Strain 원문보기

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