• Title/Summary/Keyword: 역전사효소

Search Result 113, Processing Time 0.025 seconds

Analysis of Gene Mutation and Expression Level of Follicle Stimulating Hormone Receptor in Premature Ovarian Failure(POE) Patients (조기 난소 부전증(Premature Ovarian Failure, POF) 환자에서 난포 자극 호르몬 수용체 유전자 변이 및 발현 양상에 대한 분석)

  • 김정욱;염혜원;이형송;송견지;천강우;박용석;김계현
    • Development and Reproduction
    • /
    • v.4 no.1
    • /
    • pp.61-66
    • /
    • 2000
  • This study was investigated to analyze the inactivating point mutation and expression level of follicle-stimulating hormone(FSH) receptor mRNA. In first experiment, we analyzed the point mutation. Peripheral blood was collected from each patient. To screen individuals for the C566T mutation, PCR was performed for exon 7 of the FSH receptor gene in 10 patients. No inactivating point mutation of FSH receptor gene was identified in women with premature ovarian failure. To analyze the expression level of FSH receptor, mRNA expressions were examined by RT-PCR method using specific primers for the FSH receptor. The amount of FSH receptor mRNA expressed in POF patients was lower than that in the control group. But it was not significantly different. These finding suggests that lower expression of FSH receptor in premature ovarian failure patients might be the cause of the low response to the gonadotropin during the hyperstimulation in IVF-ET cycles.

  • PDF

Evaluation of Bisphenol a Induced Apoptosis in Sertoli Cell-lines (Bisphenol A에 의한 Sertoli 세포주 내 세포자연사 검정)

  • Kim, Ji-Hyang;Kim, Jin-Kyu;Do, Byoung-Rok;Lee, Chang-Joo;Yoon, Yong-Dal
    • Development and Reproduction
    • /
    • v.9 no.2
    • /
    • pp.115-121
    • /
    • 2005
  • The present report aimed at evaluating the effect of bisphenol A(BPA) and diethylstilbestrol(DES) on Leydig or Sertoli cell-lines. To identify the differences in the susceptibility to BPA upon different cell-types, assay of the cell viability was done on TM3(Leydig cells) and TM4(Sertoli cells) cell-lines. The result indicates that Sertoli cells are more sensitive to low dose of BPA than Leydig cells. Also, the BPA- or DES-treated Sertoli cells showed a reduction of phospholipase D(PLD) activity identically. According to the confirmation of the mRNA expression of fas receptor and fas ligand in the BPA-treated cells, fas/fasL system activated by BPA will deliver the apoptosis signal onto Sertoli TM4 cells. However, Fas/FasL system was not activated in the DES-treated cells unlike the BPA-treated cells.

  • PDF

Detection of Potato Spindle Tuber Viroid Using RT-PCR Technique (RT-PCR 기법을 이용한 감자 걀쭉 바이로이드 (Potato Spindle Tuber Viroid)의 검정)

  • Joung, Young-Hee;Jeon, Jae-Heung;Choi, Kyung-Hwa;Kim, Hyun-Soon;Joung, Hyouk
    • Korean Journal Plant Pathology
    • /
    • v.13 no.4
    • /
    • pp.205-209
    • /
    • 1997
  • Potato spindle tuber viroid(PSTVd) RNAs were isolated from PSTVd-inoculated potato cv. Superioc and carried out RT-RCR with reverse transcriptase and PSTVd specifie primer pair desigened to amplify the 356 nucleotides of PSTVd genome. As a result, 356 nucleotides PCR products were amplified from PSTVd-inoculated potato cv. Superior. The 356 nucleotides DNA fragment was indeed the PSTVd geneby sequencing analysis. PSTVd could be successfully detected from infected leaf and tuber tissue of potato by using RT-PCR technique. Especially PSTVd was more effectively detected when both downstream and upstream primer were used than only downstream primer was used in RT reaction.

  • PDF

EFFECTS OF LENTINUS EDODES LECTINS ON CYTOKINE GENE EXPRESSION FROM HUMAN PERIPHERAL BLOOD MONONUCLEAR CELLS (표고버섯 렉틴의 사이토카인 생성 양상 및 특성)

  • 이인경;김희선;전경희;김성광;정시련
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 1995.04a
    • /
    • pp.113-113
    • /
    • 1995
  • 본 연구는 표고버섯에서 분리정제한 렉틴 성분(LEL)을 말초혈액 단핵세포(PBMC)에 반응시켜 사이토카인 유도능을 지질다당류(LPS)와 비교하여 역전사효소 중합반응법(RT-PCR)으로 측정하였다. 측정 대상 사이토카인은 IL-1, IL-2, IL-6, TNF$\alpha$ 및 IFN${\gamma}$의 다섯가지였으며 이들을 대상으로 PBMC에 렉틴을 적용하여 1, 8, 24, 48, 72, 96, 120분 등의 시간대에서 반응시켜 사이토카인의 유전자 발현 유도에 관한 다음의 결과를 얻었다. LEL의 사용 농도에 따른 편도선 림프구(tonsillar lymphocyte)의 TNF$\alpha$ 유전자 발현 양상은 반응 1시간의 경우 LEL의 일부 농도와 LPS 전농도에서 관찰되었으나 반응 40시간째에는 LEL 전농도와 LPS 전농도에서 TNF$\alpha$ 유전자 발현 양상을 관찰할 수 없었다. RT-PCR 결과 원액이나 회석액 재료로부터 관찰된 TNF$\alpha$유전자 band의 강 약 차이는 나타나지 않았다. LEL의 자극에 의한 반응 시간대 별 PBMC에 의한 사이토카인 유전자 발현 양상은 위에서 언급된 다섯가지 사이토카인을 유도, 생성할 수 있다는 것이 확인되었는데, IL-2, IL-6 및 IFN${\gamma}$는 120시간까지 장시간 지속되는 유전자 발현이 가능한 반면 TNF $\alpha$의 생성 양상은 이들 사이토카인의 생성 양상과는 판이하게 반응 1, 8 및 24 시간대까지만 TNF$\alpha$ 유전자 발현을 관찰할 수 있었고 IL-1은 72 시간까지 반응을 나타내는 등 특이적 양상을 보였다. 한편 LPS는 실험에 사용된 전 사이토카인의 유전자 발현을 120시간대까지 반응이 유지됨을 관찰하였기에 LPS가 PBMC의 강력한 사이토카인 유도체임을 입증할 수 있었으며 LEL과 다소 상이한 결과를 보였으나 LEL 또한 PBMC로부터 사이토카인을 생성 유지시킬 수 있는 유도체로 작용함을 확인할 수 있었다.

  • PDF

Analysis of Antigenic and Genetic Variability of G-protein of Respiratory Syncytial Virus Subgroup A Isolated in Korea over 8 Years(1990~1998) (국내에서 분리된 Respiratory Syncytial Virus A 아군의 항원성의 변이와 G-단백 mRNA의 RT-PCR 생산물의 제한효소 처리 및 염기 서열 결정을 통한 유전자 변이의 분석)

  • Choi, Eun Hwa;Park, Ki Ho;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
    • /
    • v.6 no.2
    • /
    • pp.219-233
    • /
    • 1999
  • Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

  • PDF

Outbreak of Bovine Viral Diarrhea Virus in Korean Indigenous Calf (한우송아지에서 소 바이러스성 설사병 바이러스 발생)

  • Song, Moo-Chan;Choi, Kyoung-Seong
    • Journal of Veterinary Clinics
    • /
    • v.26 no.6
    • /
    • pp.578-581
    • /
    • 2009
  • A 25-day-old, Korean indigenous calf was presented with a 10 days history of respiratory disorders and bloody diarrhea, and died. This calf was extremely unthrifty compared to others and had evidence of chronic diarrhea based on matting of feces in the hair of the tail and perineum. Ecchymotic hemorrhages were observed on multiple organs at necropsy. Bovine viral diarrhea virus (BVDV) infection was identified by RT-PCR. The phylogenetic analysis showed that this case belonged to BVDV-2a subgroup and was related to highly pathogenic USA isolate 890 (U18059). This case provided evidence for circulation of BVDV-2 in Republic of Korea. The occurrence of BVDV-2 was also reconfirmed.

Role of Chest Radiographs and CT Scans and the Application of Artificial Intelligence in Coronavirus Disease 2019 (코로나바이러스감염증 2019에서 흉부X선사진 및 CT의 역할과 인공지능의 적용)

  • Seung-Jin Yoo;Jin Mo Goo;Soon Ho Yoon
    • Journal of the Korean Society of Radiology
    • /
    • v.81 no.6
    • /
    • pp.1334-1347
    • /
    • 2020
  • Coronavirus disease (COVID-19) has threatened public health as a global pandemic. Chest CT and radiography are crucial in managing COVID-19 in addition to reverse transcription-polymerase chain reaction, which is the gold standard for COVID-19 diagnosis. This is a review of the current status of the use of chest CT and radiography in COVID-19 diagnosis and management and anㄷ introduction of early representative studies on the application of artificial intelligence to chest CT and radiography. The authors also share their experiences to provide insights into the future value of artificial intelligence.

Cloning and Structural Analysis of bfmo Operon in Methylophaga aminosulfidovorans SK1 (Methylophaga aminosulfidovorans SKI bfmo 오페론의 클로닝 및 구조 분석)

  • Lim Hyun Sook;Goo Jae Whan;Kim Lee Hyun;Kim Si Wouk;Cho Eun Hee
    • Korean Journal of Microbiology
    • /
    • v.41 no.1
    • /
    • pp.1-7
    • /
    • 2005
  • Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP) can utilize trimethylamine as a sole carbon, nitrogen, and energy source. The bacterial flavin-containing monooxygenase (bFMO) gene was identified in the strain and the recombinant enzyme expressed in E. coli oxidized trimethylamine. To study the function and regulation of the bfmo, over 8,000 nucleotide sequences of the neighboring regions including the bfmo were determined. Three open reading frames proceeding to the bfmo gene encoded analogues to highly conserved nitrate/nitrite sensing two-component system regulators and a methyl accepting protein. Two small open reading frames just downstream of the bfmo gene showed no similar proteins of known functions but the sequences were conserved among other bacteria. Reverse transcription-polymerase chain reaction analysis showed that the six putative genes consisted of three transcription units. The three regulatory genes located upstream of the bfmo gene formed two separate transcription units. The bfmo and the two downstream genes were transcribed from a single promoter.

The Effects of Hantaan Virus on the Expression of Platelet Activating Factor Receptor and on the Activity of Platelet Activating Factor Acetylhydrolase (한탄바이러스가 혈소판활성인자 수용체 발현 및 혈소판활성인자 분해효소 활성에 미치는 영향)

  • Hwang, Ji-Young;Park, Jong-Won;Hong, Sae-Yong;Park, Ho-Sun
    • Journal of Yeungnam Medical Science
    • /
    • v.25 no.1
    • /
    • pp.41-49
    • /
    • 2008
  • Background : The central physiological derangement of hemorrhagic fever with renal syndrome (HFRS) caused by hantaan virus (HTNV) is a vascular dysfunction, manifested by hemorrhage, impaired vascular tone and increased vascular permeability. Platelet activating factor (PAF), whose actions are mediated through a specific receptor, is a potent bioactive lipid. PAF has diverse biological functions in the vascular system, such as increasing vascular permeability, adhesion of leukocytes to the endothelium and reduction of cardiac output, which result in hypotension and shock. The goal of the present study was to investigate whether PAF is involved in the pathogenesis of HFRS. For this purpose, we evaluated the effect of HTNV on the expression of PAF receptor (PAF-R) and on the activity of PAF-acetylhydrolase (PAF-AH) instead of PAF because PAF is rapidly degraded by PAF-AH in vivo. Materials and methods : To evaluate the expression of PAF-R, we performed reverse-transcription PCR, western blot and FACS analyses using HTNV-infected human umbilical vein endothelial cells (HUVECs) and non-infected (control) HUVECs. In addition, we measured the activity of plasma PAF-AH in HFRS patients and normal healthy persons. Results : The mRNA and protein expression of PAF-R was increased in HTNV-infected HUVECs compared with control HUVECs at 2 and 3 days post-infection (d.p.i.). FACS analysis showed that HTNV induced the surface expression of PAF-R in HUVECs from 2 d.p.i. The activity of plasma PAF-AH was 2.5-fold lower in HFRS patients than in normal healthy persons. Conclusion : Increased PAF-R expression by HTNV might increase the responsiveness to PAF in endothelial cells. Reduced PAF-AH activity in the blood of HFRS patients might delay PAF degradation. These results suggest that changes in PAF-R and PAF-AH by HTNV might influence to PAF activity and might be involved in the vascular dysfunction of HFRS.

  • PDF

난소를 제거한 생쥐 자궁조직에서 ADAM-8, -9, -10, -12, -15, -17, -TS1의 발현

  • 김지영;배인희;이승재;최영민;김해권
    • Proceedings of the Korean Society of Developmental Biology Conference
    • /
    • 2003.10a
    • /
    • pp.59-59
    • /
    • 2003
  • ADAM은 metalloprotease/disintegrin domain을 가진 transmembrane glycoprotein으로서 지금까지 30종류 이상의 ADAM 및 10종류 이상의 ADAM-TS 단백질이 알려져 있다. 이들의 기능은 포유동물의 수정 시 sperm-egg binding과 fusion, myoblast fusion, integrin과의 결합 등에 직접 관여하거나, TNF-alpha 등의 생체신호전달물질이 세포로부터 분비될 때에 이들의 구조를 변화시켜 활성화시키는 효소로서의 작용, 그리고 dendritic cell differentiation 등에 관여하는 것으로 알려져 있다. 본 연구에서는 난소가 제거된 생쥐를 이용하여 자궁조직의 ADAM-8, -9, -10, -12, -15, -17 그리고 -TS1의 gene의 발현이 $17 \beta $-estradiol에 의하여 조절되는 지를 알아보았다. 생후 6 - 8주 된 암컷 생쥐의 난소를 제거하고, 2 주 후에 $17 \beta $-estradiol ($E_2$), progesterone ($P_4$) 혹은 이 둘 혼합액 ($E_2 + P_4$)을 sesame oil에 녹여 근육주사하였다. 2, 6, 12 시간 후 각각 자궁 조직을 얻고 유전자의 발현 양상을 알아보기 위하여 시료로부터 total RNA을 추출하여 역전사 중합효소반응 (RT-PCR)을 실시하였다. Densitometry를 이용, rpL7에 대한 ADAMS의 mRNA 발현 양을 상대적으로 분석하였다. 그 결과 ADAM-8과 -15는 6시간째에서, ADAM-10과 -TS1은 2시간째에서 sesame oil을 주사하거나 $P_4$만을 주사한 군보다 E$_2$를 주사한 군에서 mRNA의 양이 현저하게 증가하였고 ADAM-12는 2시간째에서 ADAM-17은 12시간째에서 sesame oil을 주사하거나 $P_$만을 주사한 군보다 E$_2$를 주사한 군에서 mRNA의 양이 현저하게 증가하였다. 이러한 결과로 미루어 ADAM-8, -10, -15 그리고 TS1은 progesterone에 의하여, ADAM-12와 17은 $17 \beta $-estradiol에 의하여 유전자의 발현이 upregulation 되는 것으로 생각되어진다.

  • PDF