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Usefulness of self cleavage tag mini-intein in Baculovirus Expression Vector System
Won Seok Gwak,See Nae Lee,Ji Hoon Lee,Soo Dong Woo 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.10
Recombinant proteins including a polypeptide fusion partner, termed affinity tag, to facilitate the purification of the target polypeptides are widely used. However, the affinity tag must be removed from the target after the purification process. Recently, the Synechocystis sp. PCC6803 DnaB mini-intein (Ssp DnaB mini-intein) is widely used in Escherichia coli expression systems as the solution of this problem. The Ssp DnaB mini-intein can be induced simply by shifting of pH and temperature, offering a benefit to cleave a peptide bond without using a protease or chemical reagent. Although the utility of this novel tag is widely studied in E. coli, there is no report yet in baculovirus expression vector system (BEVS). In this study, we generated several recombinant baculoviruses to express foreign proteins with Ssp DnaB mini-intein. In conclusion Ssp DnaB mini-intein was good tag also in BEVS with more advantages.
Establish a transient expression platform for the rapid development of virus-like particle vaccine
Won Seok Gwak,Hyun Soo Kim,Soo Dong Woo 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.04
The virus-like particle (VLP) is similar to a pathogenic virus and has a high immunogenicity. However, in the selection of various structural proteins to form VLP, all expression systems commonly consume most of the time and suffer from various difficulties. Therefore, there is a need for a system that can rapidly determine the structural proteins required for effective VLP production. This study aims to construct a transient expression platform using insect cells to solve this problem. Plasmid-based expression vectors and baculovirus-inducible expression vectors were constructed. The vectors were evaluated for overexpression using EGFP. We also confirmed the formation of virus like particles through overexpression of virus structural proteins.
Won Seok Gwak,Hyun Soo Kim,Jun Su Bae,Tae Hee Kim,Jin Yong Lee,Soo Dong Woo 한국응용곤충학회 2018 한국응용곤충학회 학술대회논문집 Vol.2018 No.10
Virus-like particles (VLPs) are similar to pathogenic viruses, but because they have no nucleic acid, they have excellent safety and immunogenicity and are used as a good vaccine material. However, in the selection of various structural proteins of pathogenic viruses to form VLPs, all expression systems consume a lot of time in common. Among them, the baculovirus expression system causes additional time consumption to construct the recombinant baculovirus. Therefore, there is a need for a system that can rapidly determine the structural proteins required for effective VLP production. This study aims at solving this problem by constructing a BmNPV inducible expression platform through the construction of vectors induced by BmNPV. The platform was evaluated for overexpression using EGFP. We also confirmed the formation of virus-like particles through overexpression of canine parvovirus structural proteins.
Won Seok Gwak,Soo Dong Woo 한국응용곤충학회 2019 한국응용곤충학회 학술대회논문집 Vol.2019 No.04
Among the various expression systems used for foreign protein expression, baculovirus expression system (BES) has the high level of post-translational modification ability such as glycosylation, folding and disulfide bonding. BES is widely used now in the production of VLPs because it is possible for the efficient multi-gene expression. However, there are not many cases of VLPs being manufactured through BES. Therefore, in this study, three improvements were made to increase the productivity of VLP through BES. A new heper enhanced expression vector was constructed to increase the expression of structural proteins of virus-like particles, and baculovirus bacmid was modified to increase production time. In addition, an easy purification system was constructed to efficiently purify VLP, and finally the construction of BES optimized for VLP production was completed.
Won Seok Gwak,Ji Hoon Lee,Dong Jun Kim,Hwi Geon Yun,Ji In Ma,Soo Dong Woo 한국응용곤충학회 2017 한국응용곤충학회 학술대회논문집 Vol.2017 No.10
Viral particles of Porcine epidemic diarrhea virus (PEDV) consist of a four structural proteins. Among them Spikeprotein mediated responsible for receptor binding and membrane fusion during viral infection and therefore the main targetof neutralizing antibodies. Virus-like particles (VLPs) are consisted of one or more viral structural proteins, and theirmorphologies closely resemble those of the native virus. VLPs have no virulence and can elicit robust immune responsesas compared with inactivated or live-attenuated virus vaccines. Thus, in this study, we tried two methods for VLP constructionin Bombyx mori, one is traditional method and the other is chimeric VLP method using the influenza matrix protein.Both methods could produce successfully PEDV VLPs.
Screening of anti-Nosema substances from entomopathogenic fungi by polar tube germination assay
Dong Jun Kim,Won Seok Gwak,Yong Oh Ahn,Seung Hee Lee,In Hui Kim,See Nae Lee,Dong Woo Kim,Tae Young Shin,Sung Min Bae,Soo Dong Woo 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10
The population of managed honey bees has been dramatically declining the recent past in worldwide. The one of most common disease of bees is nosemosis, the nosemosis is caused by microsporidia in the genus Nosema. Nosema apis and N. ceranae have been described as honeybee pathogens. These microsporidia are highly evoloved fungi with an obligately intracellular parasitic lifstyle. The disease causes significant detriment to honey production and results in economic losses. In our knowledge, Fumagillin is the only antibiotic approved for control of nosemosis in honey bees, however this antibiotic may have unintended effects on the honey bee host, ultimately contributing to increased prevalence and pathogenicity of Nosema. Therefore, we screened anti-Nosema substances from entomopathogenic fungal culture filtrates using in vitro polar tube germination assay. These fungal metabolites are employed as antibiotic agents. As results, Total 3 samples (23% of 13 total samples) showing the germinating inhibition against N. ceranae. This screening method may be useful for the detection of anti-Nosema substances from various samples and selected samples in this study may be a good feature to be used in the development of a new biocontrol method of nosemosis.
( Won Sohn ),( Moon Seok Choi ),( Ju Yeon Cho ),( Geum Youn Gwak ),( Yong Han Paik ),( Joon Hyeok Lee ),( Kwang Cheol Koh ),( Seung Woon Paik ),( Byung Chul Yoo ) 대한소화기학회 2014 Gut and Liver Vol.8 No.5
Background/Aims: The role of radiofrequency ablation (RFA) remains uncertain in patients with viable hepatocellular carcinoma (HCC) after transarterial chemoembolization (TACE). Methods: A total of 101 patients (April 2007 to August 2010) underwent RFA for residual or recurrent HCC after TACE. We analyzed their long-term outcomes and predictive factors. Results: The overall survival rates after RFA were 93.1%, 65.4%, and 61.0% at 1, 3, and 5 years, respectively. Predictive factors for favorable overall survival were Child- Pugh class A (hazard ratio [HR], 3.45; p=0.001), serum α-fetoprotein (AFP) level <20 ng/mL (HR, 2.90; p=0.02), and recurrent tumors after the last TACE (HR, 3.14; p=0.007). The cumulative recurrence-free survival rate after RFA at 6 months was 50.1%. Predictive factors for early recurrence (within 6 months) were serum AFP level ≥20 ng/mL (HR, 3.02; p<0.001), tumor size ≥30 mm at RFA (HR, 2.90; p=0.005), and nonresponse to the last TACE (HR, 2.13; p=0.013). Conclusions: Patients with recurrent or residual HCC who undergo prior TACE show a favorable overall survival, although their tumors seem to recur early and frequently. While good liver function, a low serum AFP level, and recurrent tumors were independent predictive factors for a favorable overall survival, poor response to TACE, a high serum AFP level, and large tumors are associated with early recurrence. (Gut Liver 2014;8:543-551)
복숭아혹진딧물 병원성 곰팡이 균주간의 살충력 차이에 대한 원인 분석
이원우, 신태영, 곽원석, 우수동 忠北大學校 農業科學硏究所 2008 農業科學硏究 Vol.36 No.2
This study was conducted to elucidate the difference of virulence between two entomopatho???? genic fungi Purpureocillium lilacinum SD17 and P. lilacinum SD13 showing high and low insecticidal activity, respectively, against the green peach aphid, Myzus persicae Sulzer. As a result of comparing and evaluating the activity of the extracellular protease and chitinase known to be the most important in insecticidal activity, the difference in activity between the two fungal isolates could not be confirmed. Accordingly, the Pr1 and Chi1 gene structures between the two fungal isolates was compared and analyzed. These genes are reported to be the most important for pathogenicity among various protease and chitinase genes. However, the difference of these genes was not also confirmed. In this study, although the reason for the difference in virulence between the two fungal isolates could not be clearly identified, it was confirmed that various studies on various factors in addition to the previously reported important factors were necessary to determine the reason of virulence. In addition to the two main factors analyzed in this study, it is expected that it will be possible to analyze the reason of the insecticidal differences between the two fungal isolates through additional analysis of various factors.