• Korean journal of applied entomology
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• v.46 no.2
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• pp.319-324
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• 2007

To determine the optimal concentration of fetal bovine serum (FBS) on the growth of insect cells and the multiplicity of viruses, the growth of cells (Sf21 and Bm5) and viruses were examined on the various concentrations of FBS. In view of the viability, growth speed, proliferation of cells and the amount of FBS, the most proper concentration for the cell culture were 7% and 5% for Sf21 and Bm5, respectively. The multiplicity of viruses at the various concentrations of FBS was similar in both cell lines at 5 days post-infection (p.i.). However, it differed significantly at 2 and 3 days p.i. The proper concentration of FBS were 10% and 3% for Sf21 at 2 and 3 days p.i., respectively, and 5% for Bm5 at both 2 and 3 days p.i. These results suggested that the optimal concentration of FBS should be determined according to the used cell lines and viruses for their optimum production.

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.53-62
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• 2012

Silkworm larvae often suffer from viral infections causing heavy losses to the economy of silk industry. Insects exhibit both humoral and cellular immune responses that are effective against various pathohens like bacteria, fungi, protozoa, etc., but no insect immune responses is effective against viral infection. To obtain genes related to insect antiviral immunity from Bombyx mori, the cDNA library was constructed from B. mori nucleopolyhedrovirus (BmNPV)-infected B. mori. From the cDNA library, we selected 411 differentially expressed clones, and the 5' ends of the inserts were sequenced to generate ESTs. In this work, 135 unigenes were generated after the assembly of 411 differentially expressed clones ESTs. Of these 135 unigenes, we selected 109 antiviral response-related candidates except 26 clones that high similarity with genes derived from BmNPV. Among 109 unigenes, a total of 80% had significant matches to genes from other organisms in the database, wheres 20% of the unigenes had not matched in the database. Functional groups of these sequences with matches in database were constructed according to their putative biological function. Three largest categories were control of cellular oraganization (52%), metabolism (20%), and protein fate (10%). The genetic information reported in this study will provide more information about antiviral-related genes in silkworms.

• Journal of Sericultural and Entomological Science
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• v.44 no.2
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• pp.69-73
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• 2002

For the practical use of nuecin protein, we tried to overexpress nuecin using Bm5 insect cell and Bombyx mori. We inserted nuecin cDNA into pBm10po1-Xa vector derived from B. mori nuclear polyhedrosis virus (BmNPV), and expressed in Bm5 cells and B. mori respectively. SDS-PAGE and Northern blot analysis showed an expressed of the protein when baculovirus expression vector system (BEVS) was used. The amount of intracellular protein is abundant, but the amount of extracellular protein is poor. The results suggest that the biologically active nuecin protein produced by using BEVS is poor because incresed level of misfolded nuecin by the strong promoter, polyhedrin and p 10 of BEVS, decrease the level of free chaperons and foldases by binding with them.

• Korean journal of applied entomology
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• v.61 no.3
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• pp.449-460
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• 2022

The morphology and whole genome sequence of Spodoptera exigua nucleopolyhedrovirus K1 (SeNPV-K1) isolated in Korea were analyzed for the use as an eco-friendly control source against S. exigua. The polyhedra of SeNPV-K1 was amorphous with a size of 0.6-1.8 ㎛, and there was no external difference with the previously reported SeNPV. As a result of analyzing the nucleotide sequence of the whole genome, it was composed of 135,756 bp, which is 145 bp more than that of the previously reported SeNPV. The G+CG+C content was 44% and there were 6 homologous repeated sequences, so there was no significant difference from the previous report. As a result of ORF analysis, SeNPV-K1 had 137, two fewer than those previously reported, and 4 ORFs present only in SeNPV-K1 were confirmed. These 4 ORFs are non-essential genes and were not considered to have a significant influence on the characteristics of the SeNPV. The genome vista analysis showed that the overall sequence similarity between SeNPV-K1 and the previously reported SeNPV was very high. The whole genome of SeNPV-K1 analyzed for the first time in Korea was found to be similar to the previously reported SeNPV, but it was confirmed that it was a novel resource in Korea with different isolate.

• Journal of Sericultural and Entomological Science
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• v.37 no.1
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• pp.46-51
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• 1995

In order to develope baculovirus expression vector system, we constructed new transfer vector of nuclear polyhedrosis virus of the silkworm, Bombyx mori. The promoter region containing only adenine of translation start codon of polyhedrin gene was cloned by polymerase chain reaction technique. And the 5' and 3' leader regions of polyhedrin gene was sequentially cloned. The polyhedrin coding gene was deleted from the ＋2 to the ＋597 position. As the result, we constructed new transfer vector which has EcoRI, SacI and KpnI sites for the cloning sites of foreign gene. New transfer vector was named as pBmKSKl. Escherichia coli $\beta$-galactosidase gene as foreign gene was inserted into pBmKSKl, under the control of the polyhedrin promoter and expressed in B. mori cells. The result showed that the new transfer vector pBmKSK1 is functional.

• Journal of Sericultural and Entomological Science
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• v.34 no.2
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• pp.20-25
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• 1992

The location of the polyhedrin gene of Bmbyx mori nuclear polyhedrosis virus(BmNPV) was determined by using a cloned polyhedrin gene from the Autographa californica nuclear polyhedrosis virus(AcNPV) as a hybridization probe. The 7.4 Kb PstⅠ fragment DNA of Bm-NPV was cloned to plasmid pUC19 vector. A fragment containing this gene was mapped and sequenced in its entire polyhedrin reading frame. Nucleotide sequences comparison of the polyhedrin of the BmNPV to that of previously reported by Ⅰatrou(1985) revealed that the sequence varied in 10 base, Comparison of the amino acid sequence of the two structured gene revealed that coding sequence varied 74 valine to isoleucine, 76 aspargine to serine and 155 methionine to valine.

• Korean Journal of Organic Agriculture
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• v.31 no.4
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• pp.395-412
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• 2023

The morphology and whole genome sequence of Hyphantria cunea nucleopolyhedrovirus W1 (HycuNPV-W1) isolated in Korea were analyzed for the use as an eco-friendly control agent against H. cunea. The HycuNPV-W1 had irregular tetrahedral polyhedra with a size of 1.5-2.2 ㎛ which is similar to that of previously reported HycuNPV isolated in Korea. As a result of whole viral genome analysis, HycuNPV-W1 was composed of 131,353 bp, which is 1,606 bp shorter than that of the previously reported HycuNPV. The G+C content was 45% and six of the homologous repeated regions were found, so there was no significant difference from the previous report. As a result of ORF analysis, HycuNPV-W1 contains total of 145 ORFs which is three ORFs less than the previous report, while two ORFs were exclusively found in HycuNPV-W1. The functions of these ORFs remains unclear and are not considered to have a significant influence on the characteristics of the HycuNPV. The genome vista analysis showed that the overall sequence identity between HycuNPV-W1 and the previously reported HycuNPV was very high. The whole genome of HycuNPV-W1 analyzed was found to be similar to those of the previously reported HycuNPV, however, it is supposed to be a novel resource in Korea with different isolate.