• The Korean Journal of Pesticide Science
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• v.18 no.3
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• pp.202-209
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• 2014

RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.2
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• pp.175-184
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• 2019

The epidermal growth factor (EGF) has a intrinsic function of inducing growth and proliferation of cells through interacting with cell membrane receptors in human epidermis and dermis layer. These functions of EGF are used as a main ingredient for wound healing medicines and anti-aging cosmetics. As a cosmetic ingredient, the EGF has a problem in exhibiting its natural efficacy due to the lack of the ability to penetrate through the stratum corneum, which is known as the skin barrier. In this study, a recombinant human epidermal growth factor ($MTD_{151}-EGF$) fused with the macromolecule transduction domain $(MTD)_{151}$ with the skin penetration ability was developed to improve the skin penetration efficiency of the EGF. Expression of $MTD_{151}-EGF$ was performed in E. coli transformed with a vector encoding the $MTD_{151}-EGF$ gene and then purified. The purified $MTD_{151}-EGF$ was evaluated using cell proliferation assay, cytotoxicity test and skin penetration test by franz diffusion cell assay and artificial skin. Cell proliferation activity of $MTD_{151}-EGF$ purified to high purity of 99% or above was equivalent to the EGF or better, and cytotoxicity was not observed. In addition, the $MTD_{151}-EGF$ showed an excellent penetration efficiency compared to the EGF in the skin penetration test with EGF and $MTD_{151}-EGF$ labeled by FITC in an artificial skin penetration model. Based on the quantitative analysis of the penetrating substance using franz diffusion cell assay, the amount of penetration was about 16 times more than that of EGF. These results can be regarded as an effective alternative to improve the existing physical transdermal penetration method related to the use of various active ingredients for cosmetics.