• Applied Biological Chemistry
• /
• 제41권3호
• /
• pp.219-223
• /
• 1998

작물 병해충에 대한 무공해 미생물 농약 개발의 일환으로 곤충 병원성 곰팡이 Metarhizium anisopliae의 분자생물학적 육종을 위해 이 균주의 형질전환계를 확립하였다. M. anisopliae의 원형질체를 제작하여 benomyl 약제에 대하여 저항성을 나타내는 Aspergillus flavus 유래의 ${\beta}-tubulin$ 유전자를 갖는 pBRG-4 plasmid DNA를 polyethylene glycol 방법으로 형질 전환하였다. 이 형질전환은 pBRG-4 DNA $50\;{\mu}g$당 10개의 효율로 이루어졌으며, 그 결과 야생형 균주들의 $2.5\;{\mu}g/ml$ 농도의 benomyl 존재 하에서도 성장이 억제되는데 반하여 선발된 형질전환체들은 $5.0\;{\mu}g/ml$ 농도의 benomyl 함유 배지에서도 잘 성장하였다. 또한 이 형질전환체들의 chromosomal DNA를 분리하여 Southern 분석한 결과 ${\beta}-tubulin$ 유전자가 homologous recombination에 의하여 M. anisopliae의 genome속에 삽입 되었음을 확인하였다.

• Mycobiology
• /
• 제44권4호
• /
• pp.325-329
• /
• 2016

A fungus, Colletotrichum fioriniae, was isolated for the first time from fruits of Chinese matrimony vine (Lycium chinense Mill.) in Korea. It was classified as C. fioriniae based on the morphological characteristics and nucleotide sequence of glyceraldehyde-3-phosphate-dehydrogenase and ${\beta}-tubulin$. To the best of our knowledge, this is the first report of C. fioriniae causing anthracnose of Chinese matrimony vine in Korea.

• 한국균학회지
• /
• 제47권1호
• /
• pp.35-42
• /
• 2019

본 연구에서는 전북 부안군에 서식하는 소나무의 침엽에서 내생균을 분리하였다. 분리된 균주는 internal transcribed spacer 영역, rDNA의 large subunit 영역, beta-tubulin 영역의 DNA 염기서열을 분석하여 동정하였다. 연구 과정에서 3종의 국내 미기록 내생균 균주를 확인하였으며, 확인된 종은 Paracamarosporiumhawaiiense, Tubakia dryina, Zasmidiumfructigenum이다. 확인된 미기록 균주의 형태적 특성 및 계통적 분석의 결과에 대해 서술하였다.

• 한국발생생물학회지:발생과생식
• /
• 제12권1호
• /
• pp.31-39
• /
• 2008

손상된 뇌신경조직내에서 신경줄기세포로부터 새로운 신경세포로의 분화가 상당히 제한되어 있어 이것이 손상된 뇌신경조직의 복구가 잘 이루어지지 않는 원인이라 여겨지고 있다. 본 연구에서는 세포배양을 통해 지방조직 중간엽 줄기세포를 도파민성 신경세포와 콜린성 신경세포로 분화를 유도하였다. 중간엽 줄기세포를 신경세포로 분화시키기 위해 N2배양액에 bFGF, EGF, dimethyl sulphoxide (DMSO)와 butylated hydroxyanisole (BHA)를 첨가하여 유도하였다. DMSO와 BHA에 처리된 중간엽 줄기세포가 빠르게 신경세포 모양으로 분화하는 것을 관찰하였으며, 이것은 면역조직학적 염색에서 신경세포 특이 표지인 $\beta$-tubulin III, 별아교세포에 대한 특이 표지인 GFAP, 흰돌기아교세포에 대한 특이 표지인 Gal-C에 대해 양성반응을 나타내었다. RT-PCR 분석에서 배양 단계에 따라 신경세포에 특이적인 표지 인자인 neuro D1, $\beta$-tubulin III, GFAP, nestin 등의 발현을 통해, 중간엽 줄기세포가 신경세포로 분화됨을 확인하였다. 그러나 중간엽줄기세포가 신경세포로 분화된 이후에는 줄기세포 표지인 SCF, C-kit와 stat-3 등은 발현되지 않았다. 또한, 중간엽줄기세포에 bFGF, SHH와 FGF8 등을 처리하면 도파민 신경세포로 분화하였다. 중간엽 줄기세포에 bFGF, RA, Shh를 처리하여 콜린성 신경세포로 분화시켰을 때, 신경세포 특이 표지인 $\beta$-tubulin III와 콜린성 신경 특이 표지인 ChAT에 양성반응를 보였다. 결론적으로 사람 지방조직의 중간엽 줄기세포가 도파민성과 콜린성 신경세포로 분화가 가능하고 이러한 잠재성을 가진 지방 유래 중간엽 줄기세포는 퇴행성 신경질환에 대한 세포 치료제로서 가능성을 제시한다.

• The Plant Pathology Journal
• /
• 제24권3호
• /
• pp.269-278
• /
• 2008

Ripe rot was frequently observed on fruits, leaves and stems of grape growing in eight locations in Korea from 2004 to 2006. All 30 isolates of Colletotrichum sp. were obtained from lesions of the ripe rot on grape plants. Out of the isolates, 19 isolates were identified as Colletotrichum acutatum and the others as Colletotrichum gloeosporioides based on morphological and cultural characteristics. Inter and intra specific variations of the Colletotrichum spp. isolates were investigated using RAPD and sequences of rDNA ITS and $\beta$-tubulin-2. Isolates of C. acutatum and C. gloeosporioides were distinctly differentiated by molecular analyses. Phylogenetic trees of ITS and$\beta$-tubulin-2 showed that Korean isolates of C. acutatum were clustered into groups A2 and A3 among the eight global groups. A2 included non-chromogenic isolates and A3 chromogenic ones. Both C. acutatum and C. gloeosporioides isolates were tested for pathogenicity to grape leaves. All isolates tested induced lesions on the leaves of grape by artificial inoculation. There was no difference in pathogenicity between C. acutatum and C. gloeosporioides isolates. This is the first report that C. acutatum except C. gloeosporioides causes grape ripe rot in Korea.

• Korean Journal of Acupuncture
• /
• 제29권2호
• /
• pp.260-270
• /
• 2012

Objectives : Hippocampus, a region of temporal lobe, plays an important role in the pathogenic mechanisms of brain diseases such as Alzheimer's disease, depression and temporal lobe epilepsy. This research is designed to investigate hippocampal changes after acupuncture stimulation at Shinmun(HT7) using 2-dimensional gel electrophoresis(2-DE). Methods : On postnatal-day 15, rat pups were randomly devided into Normal(NOR) or HT7 group. All of Pups kept with their mothers for 7 days, but pups in HT7 group received acupuncture stimulation at HT7 daily. On postnatal-day 21, hippocampus of each rat pup was dissceted 30 minutes after last acupuncture stimulation and the protein expressions were investigated using 2-DE. Results : After acupuncture stimulation at HT7, expression of 20 proteins were significantly increased. Succinate semialdehyde dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase-like, transketolase, aconitate hydratase and phosphoglucomutase-1 were related to glucose methabolism. Eukaryotic initiation factor(eIF) 4A-II, eIF 4A-III, mitochondrial Tu translation elongation factor and chain A of crystal structure of the 70-Kda heat shock cognate protein involve in the protein synthesis in ribosome. Tubulin ${\beta}$-4 chain, tubulin T ${\beta}$-15 and tubulin ${\alpha}$-1B chain comprise cytoskeleton. Glutathione S-transferase(GST) ${\omega}$-1, GST P and GST Yb-3 can reduce oxidative stress. ${\beta}$-soluble N-ethylmaleimide-sensitive fusion protein attachment protein is required for vesicular transport between the endoplasmic reticulum and the Golgi apparatus, glycerol-3-phosphate dehydrogenase plays a major role in lipid biosynthesis, creatine kinase U-type catalyses the conversion of creatine and consumes adenosine triphosphate to create phosphocreatine and adenosine diphosphate. Platelet-activating factor acetylhydrolase IB subunit alpha and voltage depedent anion-selective channel protein 2 were also increased. Conclusions : The results suggest that acupuncture stimulation at HT7 may enhance glucose and lipid metabolism, protein synthesis, cytoskeletal substance and anti-oxidative stress in hippocampus.

• BMB Reports
• /
• 제41권3호
• /
• pp.210-216
• /
• 2008

CSE1L/CAS, a microtubule-associated, cellular apoptosis susceptibility protein, is highly expressed in various cancers. Microtubules are the target of paclitaxel-induced apoptosis. We studied the effects of increased or reduced CAS expression on cancer cell apoptosis induced by chemotherapeutic drugs including paclitaxel. Our results showed that CAS overexpression enhanced apoptosis induced by doxorubicin, 5-fluorour-acil, cisplatin, and tamoxifen, but inhibited paclitaxel-induced apoptosis of cancer cells. Reductions in CAS produced opposite results. CAS overexpression enhanced p53 accumulation induced by doxorubicin, 5-fluorouracil, cisplatin, tamoxifen, and etoposide. CAS was associated with $\alpha$-tubulin and $\beta$-tubulin and enhanced the association between $\alpha$-tubulin and $\beta$-tubulin. Paclitaxel can induce G2/M phase cell cycle arrest and microtubule aster formation during apoptosis induction, but CAS overexpression reduced paclitaxel-induced G2/M phase cell cycle arrest and microtubule aster formation. Our results indicate that CAS may play an important role in regulating the cytotoxicities of chemotherapeutic drugs used in cancer chemotherapy against cancer cells.

• Journal of Korean Neurosurgical Society
• /
• 제44권6호
• /
• pp.375-381
• /
• 2008

Objective : The effects on neural proliferation and differentiation of neural stem cells (NSC) of basic fibroblast growth factor-2 (bFGF). insulin growth factor-I (IGF-I). brain-derived neurotrophic factor (BDNF). and nerve growth factor (NGF) were assessed. Also, following combinations of various factors were investigated : bFGF+IGF-I, bFGF+BDNF, bFGF+NGF, IGF-I+BDNF, IGF-I+NGF, and BDNF+NGF. Methods : Isolated NSC of Fisher 344 rats were cultured with individual growth factors, combinations of factors, and no growth factor (control) for 14 days. A proportion of neurons was analyzed using $\beta$-tubulin III and NeuN as neural markers. Results : Neural differentiations in the presence of individual growth factors for $\beta$-tubulin III-positive cells were : BDNF, 35.3%; IGF-I, 30.9%; bFGF, 18.1%; and NGF, 15.1%, and for NeuN-positive cells was : BDNF, 34.3%; bFGF, 32.2%; IGF-I, 26.6%; and NGF, 24.9%. However, neural differentiations in the absence of growth factor was only 2.6% for $\beta$-tubulin III and 3.1% for NeuN. For $\beta$-tubulin III-positive cells, neural differentiations were evident for the growth factor combinations as follows : bFGF+IGF-I, 73.1 %; bFGF+NGF, 65.4%; bFGF+BDNF, 58.7%; BDNF+IGF-I, 52.2%; NGF+IGF-I, 40.6%; and BDNF+NGF, 40.0%. For NeuN-positive cells : bFGF+IGF-I, 81.9%; bFGF+NGF, 63.5%; bFGF+BDNF, 62.8%; NGF+IGF-I, 62.3%; BDNF+NGF, 56.3%; and BDNF+IGF-I, 46.0%. Significant differences in neural differentiation were evident for single growth factor and combination of growth factors respectively (p<0.05). Conclusion : Combinations of growth factors have an additive effect on neural differentiation. The most prominent neural differentiation results from growth factor combinations involving bFGF and IGF-I. These findings suggest that the combination of a mitogenic action of bFGF and post-mitotic differentiation action of IGF-I synergistically affects neural proliferation and NSC differentiation.

• 한국식품저장유통학회지
• /
• 제15권1호
• /
• pp.84-87
• /
• 2008

유전자 변형 작물은 생산성 측면에서 많은 장점이 있지만 이를 섭취할 경우 잠재적인 위험 요소들에 의해 많은 문제가 대두대고 있다. 본 연구는 저항성유전자를 이입한 배추에서 Profillin, Tubulin-${\alpha}$ (Tub-${\alpha}1$), Heat-shock protein (Bchsp 17.6) and Ubiquitin conjugating enzyme (UBE)의 발현과 이를 30일간 섭취한 마우스에서 ${\beta}$-actin(${\beta}$-act), ${\beta}$-2-microglobulin (B2m), Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and ${\beta}$-glucuronidase (Gus)의 발현 정도를 RT-PCR을 통해 알아보았다. 실험 결과 저항성유전자를 이입한 배추와 그렇지 않은 배추의 유전자 발현 패턴은 큰 차이를 보이지 앓았으며, 이를 섭취한 마우스 장기에서도 발현에 따른 큰 차이는 나타나지 않았다.

• The Plant Pathology Journal
• /
• 제25권4호
• /
• pp.322-327
• /
• 2009

A total of 23 isolates of Botrytis cinerea causing the grey mold were collected from infected ginseng in several fields of Korea. The sensitivity to carbendazim and the mixture of carbendazim plus diethofencarb was determined through a mycelial inhibition test on PDA amended with or without fungicides. B. cinerea isolates were classified as 3 phenotypes, which were the first phenotype resistant to both of carbendazim and the mixture ($Car^RMix^R$), the second one resistant to carbendazim and sensitive to the mixture ($Car^RMix^S$), and the last one sensitive to both of them ($Car^RMix^S$). Carbendazim resistance correlated with a single mutation $\beta$-tubulin gene of B. cinerea amplified with primer pair tubkjhL and tubkjhR causing a change of glutamate to alanine at amino acid position 198. Furthermore, the substitution of valine for glutamate led the resistance to carbendazim and the mixture at the same position of amino acid. PCR-restriction fragment length polymorphism (PCR-RFLP) analysis using the restriction endonuclease, Tsp451 and BstUI allowed differentiation of the PCR fragment of $\beta$-tubulin gene of $Car^SMix^S$ isolates from that of $Car^RMix^R$ and $Car^RMix^S$ isolates. This method will aid in a fast detection of resistance of carbendazim and the mixture of carbendazim plus diethofencarb in B. cinerea in ginseng field.