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유영법(Young-Beob Yu),Mi-Jung Kim(김미정),Do-Young Yum(염도영),Bon-Taq Koo(구본탁),Hye-Kyeong Ha(하혜경),Hyun-Kyu Shin(신현규),Jin-Yeul Ma(마진열),Jong-Cheol Park 한국한의학연구원 2006 한국한의학연구원 연구보고서 Vol.- No.-
The purpose of this study was to examine the effects of the Hominis placenta extracts and Astragali radix extracts on improvement of senile osteoporosis (Type 2) in SAM P6. Placenta ex-tracts (Beuronel<SUP>®</SUP>) produced by INBIONET Corp, Daejeon, Korea is hydrolysate of dried human placenta that contains interferon, prostaglandin E1, lysozyme, glycosaminoglycans (GAG) and various amino acid such as leusine, alanine and glutamic acid. In traditional oriental medicine, Placenta (Jahageo) is indicated for insufficiency of blood and tidal fever due to steaming bone disorder. At 10 weeks after birth, Placenta ex. and Astrali Radix ex. was given 100μL and 200μL/kg/day (i.p.), 3times a week for 4 weeks in SAM P6 mice. We measured complete blood cells (CBC) such as RBC, HGB, Hct, PLT, MPB and MCHC. And we analyzed the plasma concentrations of blood urea nitrogen, creatinine, inorganic phosphate and total iron. In addition, we tested bone mineral density (BMD) using the soft X-ray (Lunar Pixirnus). As a result, in SAM P6, injection of Placenta ex. (200μL) increased in RBC, HB and PLT, in comparison with control group. It was also found that the inorganic phosphate levels increased significantly in the injection groups of the Placenta ex. from that of the control group, but blood urea nitrogen was no significant. Moreover, Placenta ex. and Astrali Radix ex. were showed a trend of increase in bone mineral density (BMD) of the vertebrae lumbales, femur and tibia in P6 mice (Fig. 1). These findings suggest that Placenta ex. and Astragali Radix ex. are effective in preventing bone loss in SAMP6. In conclusion, these extracts help on improvement of osteoporosis in SAMs through probably hematogenous functions. <그림참조>
Properties of Recombinant Derivatives of pJY501, A Multi-copy Streptomyces plasmid
Yum, Do Young,Kong, In Soo,Yu, Ju Hyun 한국산업미생물학회 1990 한국미생물·생명공학회지 Vol.18 No.1
Thiostrepton 내성 유전자(tsr)를 포함하는 multicopy 재조합 플라스미드 pJY502(5.5kb)의 제한효소 지도를 비교해본 결과 pJY502는 새로운 플라스미드로 확인되었다. pJY502는 Streptomyces에서 넓은 host range를 나타내었으며 cloning에 사용할 수 있는 단일 BglⅡ 제한효소 인식부위를 갖고 있었다. pJY502의 형질전환 빈도는 S. lividans에서 2.2×10 exp(5)이었다. 또한 E. coli-Streptomyces bifunctional 플라스미드 pJY504을 제조하였다. The restriction cleavage map of multi-copy recombinant plasmid, pJY502(5.5kb), carrying the thiostrepton resistance gene(tsr) was determined. Comparison of the restriction pattern with that of Streptomyces plasmids previously demonstrated that pJY502 was novel. The plasmid pJY502 had a broad host range in Streptomyces and contained single BglⅡ site for cloning purpose. Transformation frequency of pJY502 was 2.2×10 exp(5) in S. lividans. E. coli-Streptomyces bifunctional plasmid, pJY504, was also constructed.
호알칼리성 Bacillus sp.가 생산하는 Bacteriolytic Enzyme을 이용한 Bacillus subtilis의 형질전환
유주현,이인숙,옥승호,박희경,염도영,배동훈 한국산업미생물학회 1993 한국미생물·생명공학회지 Vol.21 No.5
호알칼리성 Bacillus sp. YJ-451이 생산하는 bacteriolytic enzyme은 Bacillus subtilis 원형질체 형질전환에 사용되고 있는 세포벽 용해효소 lysozyme과 그 작용기작이 서로 다른 endopeptidase로 peptidoglycan peptide subunit의 L-alanine과 D-glutarnic acid 사이를 절단한다. 이 lytic enzyme을 이용하여 B. subtilis의 원형질체 형질전환에 대한 조건을 검토하였다. 대수기 말기까지 생육시킨 균체에 lytic enzyme을 5×10exp(2) U/㎖ 농도로 37℃ 에서 pH 6.5로 90분간 처리하였을 때 원형질체 형성이 가장 잘 되었다. 원형질체 형성을 위한 완충용액내의 osmotic stabilizer인 sucrose는 0.5 M 농도가 가장 적당하였으며 0.8% agar가 첨가된 pH 7.3의 DM-3 재생배지에서 37℃ 로 재생시켰을 때 재생율이 가장 높았다. 형질전환 빈도는 30%(w/v) PEG 6000으로 2분간 처리하였을 때 최고의 형질전환 빈도를 나타내었다. B. subtilis 형질전환에 사용된 pUB110의 농도에 따른 형질전환율은 직선적으로 비례하는 양상을 보였으며, lytic enzyme을 사용하여 형질전환하였을 경우 lysozyme을 사용한 경우보다 형질전환 빈도는 다소 낮았으나 재생시간은 약 30시간 이상 빠르게 일어나 신속한 형질전환에 유리하였다. The extracellular bacteriolytic enzyme from alkalophilic Bacillus sp. YJ-451 was endopeptidase which hydrolyzes the peptide bond at the amino group of D-glutamic acid in the peptidoglycan. Protoplast transformation system of B. subtilis by the lytic enzyme that differs, in mechanisms, from lysozyme which was used to transformation of B. subtilis was investigated. High protoplast yield was obtained from cells cultured in PAB at the late logarithmic growth phase. Protoplasts of B. subtilis were obtained at best efficiency by treatment with 5×10exp(2) U/㎖ lytic enzyme in the pH 6.5 of SMMP buffer containing 0.5 M sucrose. Cell wall was regenerated efficiently on DM-3 (pH 7.3) medium containing 0.8% agar. Under the best condition for protoplast formation and regeneration the highest transformation efficiency was achieved with 30% (w/v) PEG (M.W. 6000) treatment for 2 min. The transformation efficiency according to plasmid DNA concentration showed a linear relationship to DNA amounts. The transformation frequency by lytic enzyme was lower than lysozyme, but the regeneration time of the one was more rapid by 30 hr than the other.