Transcription and Expression of Major VP Gene of Bombyx Mori Parvo-like Virus

Jinsong Cheng, Qin Yao, Meng Lv, Chen Sun, Yuanqing He, Keping Chen

Abstract


The Bombyx mori parvo-like virus (China Zhenjiang isolate), termed as BmDNV-Z, replicates only in host’s midgut
columnar cells and causes flacherie disease. The viral genome is composed of two sets of different single-stranded
linear DNA molecules, Viral DNA 1(VD1) and Viral DNA 2(VD2). Here we cloned the major VP gene from the
plasmid contained viral DNA fragments. The gene consists of 1500 nucleotides and the deduced protein has 499 amino
acid residues, with the predicted molecular weight of 54934.24Da, isoelectric point of 6.73. The coding sequence was
inserted into expression plasmid vector pET-30a and expressed in Escherichia coli BL21 (DE3) under the inducible
promoter LacZ. The expressed product was detected by Western blotting with anti-His antibody. Interestingly, there are
four laddered band instead of one. We speculated that this virus may use leaky scanning mechanism to express four
proteins in one ORF. And we investigated the major VP gene transcriptional time phase in susceptible silkworm through
real time quantitative PCR (qPCR). The result showed that the mRNA level of VP gene increased greatly after 24 hours
post inoculation.

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International Journal of Biology   ISSN 1916-9671(Print)   ISSN 1916-968X  (Online)

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