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Peroxidase mRNA 의 In vitro Translation 및 특성에 관한 연구
김재종,김승수 ( Jae Jong Kim,Soung Soo Kim ) 생화학분자생물학회 1987 BMB Reports Vol.20 No.3
Total celluar RNA was isolated from tobacco callus (Nicotiana tabacum L., var Virginia 115) with guanidium thiocynate and guanidine HCl method. The total RNA was further fractionated by oligo (dT)-cellulose chromatography. For translation of peroxidase mRNAs in cell-free protein-synthesizing system, rabbit reticulocyte lysate was tested and the optimum translational conditions of peroxidase mRNAs were established. The lysate containing 2 mM Mg^(2+), 80 mM K^+ and 15 μM hemine was found to be the optimum condition for the translation of one population of peroxidase mRNAs. On the other hand, 1 mM Mg^(2+) was the optimum for the other population of peroxidase mRNAs. The incorporation of [^(35)S]-Met into peroxidase was increased up to 1 ㎍ of poly(A)^+ RNA and then it was decreased. Size fractionation of total cellular RNA was carried out by 6M urea agarose electrophoresis. Peroxidase mRNAs were distributed at slightly above 16S and 23S regions. As a result of immunoprecipitation of in vitro translation products with anti isoperoxidase C₃ and A₄ antiserum, we found C₃, C₁ or/and C₂, C₄ crossreactive to anti C₃ antiserum and A₄, A₁ to anti A₄ antiserum.
In Vitro Translation and Characterization of Peroxidase mRNAs from Tobacco Callus
김재종,김승수,Kim, Jae-Jong,Kim, Soung-Soo 생화학분자생물학회 1987 한국생화학회지 Vol.20 No.3
담배 callus (Nicotiana tabacum L., var Virginia 115)로 부터 guanidium thiocyanate 및 guanidine HCl 방법을 이용 total cellular RNA를 분리 하였으며 oligo (dT)-cellulose 친화성 크로마토그라피 방법에 의해 $poly(A)^+$ RNA를 분리하였다. Protein-synthesizing cell-free system을 이용한 peroxidase mRNA의 translation을 위 해 rabbit reticulocyte lysate의 peroxidase mRNA에 대한 최적 translation 조건을 조사한 결과 2mM $Mg^{2+}$, 80 mM $K^+$, $15\;{\mu}M$ hemine 및 $1\;{\mu}g$의 $poly(A)^+$ RNA 일때 최대의 방사능 유입과 peroxidase 활성을 나타냈다. 그러나 어떤 peroxidase population에 있어서는 1 mM $Mg^{2+}$을 최적 조건으로 갖는 경우도 있었다. 6M urea agarose electrophoresis에 의해 total cellular RNA를 분석해 본 결과 16S 및 23S 부근에서 peroxidase mRNA를 확인할 수 있었다. 또한 Anti isoperoxidase $C_3$ 및 $A_4$antiserum을 이용한 immunoprecipitation에 의해 in vitro translation products 내에서 anti $C_3$와 crossreactivity를 갖는 isoperoxidase $C_3$, $C_4$, $C_1/C_2$ 및 anti $A_4$와 crossreactivity를 갖는 isoperoxidase $A_4$, $A_1$을 확인할 수 있었다. Total celluar RNA was isolated from tobacco callus (Nicotiana tabacum L., var Virginia 115) with guanidium thiocynate and guanidine HCl method. The total RNA was further fractionated by oligo (dT)-cellulose chromatography. For translation of peroxidase mRNAs in cell-free protein-synthesizing system, rabbit reticulocyte lysate was tested and the optimum translational conditions of peroxidase mRNAs were established. The lysate containing 2 mM $Mg^{2+}$, 80 mM $ K^+$ and $15\;{\mu}M$ hemine was found to be the optimum condition for the translation of one population of peroxidase mRNAs. On the other hand, 1 mM $Mg^{2+}$ was the optimum for the other population of peroxidase mRNAs. The incorporation of [$^{35}S$]-Met into peroxidase was increased up to $1\;{\mu}g$ of $poly(A)^+$ RNA and then it was decreased. Size fractionation of total cellular RNA was carried out by 6M urea agarose electrophoresis. Peroxidase mRNAs were distributed at slightly above 16S and 23S regions. As a result of immunoprecipitation of in vitro translation products with anti isoperoxidase $C_3$ and $A_4$ antiserum, we found $C_3$, $C_1$ or/and $C_2$, $C_4$ crossreactive to anti $C_3$ antiserum and $A_4$, $A_1$ to anti $A_4$antiserum.
메티실린 내성 황색 포도상 구균에서 mecA, femA 유전자의 임상적 의의
박정은,김택선,박수성,김은령,김일수,안일영,김영진,김재종,강성옥,박한오,Park, Jung-Eun,Kim, Taek-Sun,Park, Su-Sung,Kim, Eun-Ryoung,Kim, Il-Su,Ann, Il-Young,Kim, Young-Jin,Kim, Jae-Jong,Kang, Sung-Ok,Park, Han-Ho 대한소아감염학회 1996 Pediatric Infection and Vaccine Vol.3 No.2
Purpose : In the treatment of MRSA infection, rapid detection of MRSA is extremely important. The mecA gene codes the new drug resistant polypeptides called PBP2' which mediates the clinically relevant resistance to all beta-lactam antibiotics. The identical mecA gene has been found in coagulase-negative staphylococcus with the methicillin-resistant phenotype. On the other hand, the femA gene was absent from coagulase negative staphylococcus strains with the methicillin resistant phenotype. This study is aimed at early detection and definite diagnosis of MRSA. Methods : A total of 24 MRSA strains were studied. All strains were tested for antimicrobial susceptibility and purified DNA. We amplified both mecA and femA genes by PCR in 24 strains. Results : In MRSA all the 16 strains (100%) carried femA gene and 11 strains (68.7%) carried mecA gene. In contrast, in methicillin sensitive staphylococcus all the 8 strains (100%) carried femA and only 3 strains (37.5%) were detected mecA. Conclusions : As results, there are difference in the phenotype and genotype of methicillin resistance by PCR of mecA and femA. Such disparities between methicillin resistance and the presence of mecA gene suggest the presence of control gene of the mecA.
한국인 집단에서 알츠하이머 치매의 임상적 진단을 위한 ApoE 유전자와 BchE-k 변이형과의 상승관계에 관한 연구
신은심 ( Eun Sim Shin ),정호진 ( Ho Jin Choung ),장미선 ( Mi Sun Chang ),윤송로 ( Song Ro Yoon ),임용빈 ( Yong Bin Eym ),이기철 ( Gi Chul Lee ),이강희 ( Kang Hee Lee ),김재종 ( Jae Jong Kim ),최수경 ( Soo Kyung Choi ) 대한임상검사과학회 1999 대한임상검사과학회지(KJCLS) Vol.31 No.2
The Apolipoprotein E type 4 allele (ApoE e4) is genetically associated with the common late onset familial and sporadic forms of Alzheimer``s disease. ApoE gene is located on chromosome 19, and the three common alleles are designated e 2, e 3, and e 4. The e 4 gene is a major risk factor for late onset Alzheimer``s disease(LOAD). It has been reported that the polymorphic k variant of butyrylcholinesterase (BchE-k) has an elevated frequency in AD patients carrying the e 4 allele of ApoE when compared with a nonnal hea1thy group. BchE activiη in the brain increases around age 60 and is elevated in AD. The BchE-k variant, which is the common variant of the BchE gene, has been reported to show allelic association with AD in subjects who are also carriers of the e 4 allele of ApoE, especilly in subjects over the age of 75. This study was performed to evaluate the distribution of ApoE and BchE genotype in healthy and AD group and to evaluate the synergy between BchE-k variant and ApoE з 4 in AD. ApoE and BchE genotype were determined in DNA samples from 610 healthy group and 60 LOAD patients by using ARMS a11ele specific oligonucleotides (ASOs) amplification by standard agarose gel electrophoresis. πle efIect of ApoE E з4 was significant1y associated with AD(p<0.05). A comparison between AD patients and the hea1thy individua1s, both with the E 4 a11ele, indicated an interaction between BchE-k and ApoE E з4(p<0.05). The association of BchE-k with AD was limited to carriers of ApoE E 4 a11ele, among whom the presence of BchE-k gave an odds ratio of AD of 3.48 (95% C.I. 1.3-9.2). Together these resu1ts suggested that further evidence of an association between ApoE E 4 and LOAD, and BchE-k acts in synergy with ApoE E з4 as a susceptibility gene for AD.
항정신병약물을 투여 받은 정신분열병환자의 체중,BMI,및 혈장 Leptin 농도의 변화
김재종,변기욱,김봉조,손진욱,박철수 대한생물치료정신의학회 2000 생물치료정신의학 Vol.6 No.1
Objective : Weight gain is a major side elect of treatment with newer antipsychotics. The mechanism, however, of weight gain is poorly understood. Recent researches suggested that there is a positive relationship between the levels of plasma leptin and the weight gains of the schizophrenic patients receiving the antipsychotics. This study was conducted to verify, firstly, the effect of the use of olanzapine and haloperidol on the weight gain, BMI, and the change of plasma leptin concentration and to find, secondly, whether there is any co-relationship between the plasma leptin level and the antipsychotic-induced weight gain. Method : 18 male schizophrenia inpatients were divided into two groups receiving olanzapine(N=8) or haloperidol(N=10) treatment and weight, BMI and plasma leptin level were measured weely over 5 weeks. Prior to the whole measurements, and patients were curtailed all diet from 21:00 of the previous day and the blood samples were collected at 8:00 and 8:15 in the morning. Results : In schizophrenic patients receiving olanzapine, significant increases in weight and BMI were found from the second week of medication, while those of the plasma leptin levels were shown from third week. In schizophrenic patients receiving haloperidol, no significant increases in the measurements were found for the 5 weeks of medication period. There was a close co-relationship between the plasma leptin concentration and the body weight increases. The weight gains were initiated from the second week of medication while the increases of plasma leptin concentration were started from the third week. The weight gains were followed by the continuative increases in plasma leptin levels for up to 5 weeks. Conclusion : Medication of olanzapine appears to be associated with an increase in weight, BMI, and plasma leptin level, whereas that of haloperidol does not. From the results observed the following suggestion could be made that most probable reason for olanzapine-induced increases in leptin levels are weight gain.