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101.
Yeon-Gu Kim Byoungwoo Park Jung Oh Ahn Joon-Ki Jung Hong Weon Lee Eun Gyo Lee 《BMC biotechnology》2012,12(1):1-6
Background
The establishment of high producer is an important issue in Chinese hamster ovary (CHO) cell culture considering increased heterogeneity by the random integration of a transfected foreign gene and the altered position of the integrated gene. Fluorescence-activated cell sorting (FACS)-based cell line development is an efficient strategy for the selection of CHO cells in high therapeutic protein production.Results
An internal ribosome entry site (IRES) was introduced for using two green fluorescence protein (GFP) fragments as a reporter to both antibody chains, the heavy chain and the light chain. The cells co-transfected with two GFP fragments showed the emission of green fluorescence by the reconstitution of split GFP. The FACS-sorted pool with GFP expression had a higher specific antibody productivity (q Ab) than that of the unsorted pool. The q Ab was highly correlated with the fluorescence intensity with a high correlation coefficient, evidenced from the analysis of median GFP and q Ab in individual selected clones.Conclusions
This study proved that the fragment complementation for split GFP could be an efficient indication for antibody production on the basis of high correlation of q Ab with reconstitution of GFP. Taken together, we developed an efficient FACS-based screening method for high antibody-producing CHO cells with the benefits of the split GFP system. 相似文献102.
Sin‐Ae Lee Tae You Kim Tae Kyoung Kwak Hyeonjung Kim Semi Kim Hyo Jeong Lee Sung‐Hoon Kim Ki Hun Park Hyun Jeong Kim Moonjae Cho Jung Weon Lee 《Journal of cellular biochemistry》2010,111(1):59-66
Overexpression of transmembrane 4 L six family member 5 (TM4SF5), a four‐transmembrane L6 family member, causes aberrant cell proliferation and angiogenesis, but the roles of TM4SF5 in migration, invasion, and tumor metastasis remain unknown. Using in vitro hepatocarcinoma cells that ectopically or endogenously express TM4SF5 and in vivo mouse systems, roles of TM4SF5 in metastatic potentials were examined. We found that TM4SF5 expression facilitated migration, invadopodia formation, MMP activation, invasion, and eventually lung metastasis in nude mice, but suppression of TM4SF5 with its shRNA blocked the effects. Altogether, TM4SF5‐mediated migration and invasion suggest that TM4SF5 may be therapeutically targeted to deal with TM4SF5‐mediated hepatocellular cancers. J. Cell. Biochem. 111: 59–66, 2010. © 2010 Wiley‐Liss, Inc. 相似文献
103.
Santhanam R Okoro CK Rong X Huang Y Bull AT Andrews BA Asenjo JA Weon HY Goodfellow M 《Antonie van Leeuwenhoek》2012,101(3):575-581
The taxonomic position of a Streptomyces strain isolated from a hyper-arid desert soil was established using a polyphasic approach. The organism had chemical and
morphological properties typical of the genus Streptomyces and formed a phyletic line at the periphery of the Streptomyces coeruleorubidus subcluster in the 16S rRNA gene tree. DNA:DNA relatedness values between the isolate and its nearest phylogenetic neighbours,
Streptomyces lomondensis NRRL 3252T and Streptomyces lusitanus NRRL B-12501T were 42.5 (±0.48)% and 25.0 (±1.78)%, respectively. The isolate was readily distinguished from these organisms using a combination
of morphological and phenotypic properties. On the basis of these results, it is proposed that isolate C63T (CGMCC 4.6997T, = KACC 15425T) be classified as the type strain of Streptomyces deserti sp. nov. 相似文献
104.
F-box proteins, components of the Skp1-Cullin1-F-box (SCF) protein E3 ubiquitin ligase complex, serve as the variable component responsible for substrate recognition and recruitment in SCF-mediated proteolysis. F-box proteins interact with Skp1 through the F-box motif and with ubiquitination substrates through C-terminal protein interaction domains. F-box proteins regulate plant development, various hormonal signal transduction processes, circadian rhythm, and cell cycle control. We isolated an F-box protein gene from wheat spikes at the onset of flowering. The Triticum aestivum cyclin F-box domain (TaCFBD) gene showed elevated expression levels during early inflorescence development and under cold stress treatment. TaCFBD green fluorescent protein signals were localized in the cytoplasm and plasma membrane. We used yeast two-hybrid screening to identify proteins that potentially interact with TaCFBD. Fructose bisphosphate aldolase, aspartic protease, VHS, glycine-rich RNA-binding protein, and the 26S proteasome non-ATPase regulatory subunit were positive candidate proteins. The bimolecular fluorescence complementation assay revealed the interaction of TaCFBD with partner proteins in the plasma membranes of tobacco cells. Our results suggest that the TaCFBD protein acts as an adaptor between target substrates and the SCF complex and provides substrate specificity to the SCF of ubiquitin ligase complexes. 相似文献
105.
Since 1957, Microsporum (M.) canis has been one of the common causative agents of dermatophytosis in Korea. We analyzed 944 patients infected with M. canis who attended outpatient clinic over a 17-year period (1993–2009). M. canis infections were diagnosed by mycological examinations, including direct microscopic examinations with 15% KOH and cultures
on potato dextrose agar complemented with 0.5% chloramphenicol. Mycological examinations confirmed 164,903 cases of dermatophytosis,
944 (0.6%) of which were M. canis infections. The annual prevalence of patients with M. canis infection was the highest in 2002 (91 cases). Then, the prevalence of patients with M. canis infection markedly decreased until 2008 (15 cases). The ratio of male to female patients was 0.65:1, but the ratio in children
under the age of 15 was 1.14:1 and was 0.20:1 in adults. Seasonally, 274 cases occurred in winter, showing higher incidence
than in other seasons. There was a difference in the clinical forms of M. canis infections between children and adults; tinea (T.) capitis was the most common form in children, but T. corporis was the
most common in adults. We demonstrate that the decreasing prevalence of M. canis infections over the study period has been noted in Korea. 相似文献
106.
Yong Weon Yi Wooyoung Hong Hyo Jin Kang Hee Jeong Kim Wenjing Zhao Antai Wang Yeon‐Sun Seong Insoo Bae 《Journal of cellular and molecular medicine》2013,17(5):648-656
Triple‐negative breast cancers (TNBCs) are known to be intrinsically resistant to inhibitors for epidermal growth factor receptor (EGFR). Until now, clinical trials for TNBCs using EGFR inhibitors (EGFRis) as single agents have yielded disappointing results. Here, we report that combinatorial treatment using EGFRis, such as gefitinib or erlotinib, with PI3K/AKT pathway inhibitors (PI3K/AKTis) demonstrated a synergistic, anti‐proliferative effect in cell lines of the basal‐like (BL) subtype, a subtype of TNBC. Western blot analysis revealed that the gefitinib/PI‐103 combination significantly reduced the level of both phospho‐AKT and phospho‐ERK in two susceptible BL subtype cell lines, SUM149PT and MDA‐MB‐468, whereas it had little or no effect on the level of phospho‐ERK in two non‐susceptible cell lines (HS578T and MDA‐MB‐231) of mesenchymal stem‐like (MSL) TNBC subtype. The gefitinib/PI‐103 combination also significantly induced caspase‐3/7‐mediated PARP cleavage and reduced two anti‐apoptotic proteins, XIAP and Bcl‐2 in the susceptible cell lines. In addition, the level of myeloid cell leukemia 1 (Mcl‐1) protein was markedly decreased by gefitinib/PI‐103 combination in the BL TNBC cells, but showed no significant change by this combination in MSL subtype cells. These results suggest that pharmacological inhibition of EGFR used in combination of PI3K/AKTis is a potential therapeutic approach to treat a subtype of TNBCs. 相似文献
107.
Hong‐Quan Duong Young Bin Hong Jung Soon Kim Hee‐Seok Lee Yong Weon Yi Yeon Jeong Kim Antai Wang Wenjing Zhao Chi Heum Cho Yeon‐Sun Seong Insoo Bae 《Journal of cellular and molecular medicine》2013,17(10):1261-1270
Checkpoint kinase 2 (CHK2) plays pivotal function as an effector of cell cycle checkpoint arrest following DNA damage. Recently, we found that co‐treatment of NSC109555 (a potent and selective CHK2 inhibitor) potentiated the cytotoxic effect of gemcitabine (GEM) in pancreatic cancer MIA PaCa‐2 cells. Here, we further examined whether NSC109555 could enhance the antitumour effect of GEM in pancreatic adenocarcinoma cell lines. In this study, the combination treatment of NSC109555 plus GEM demonstrated strong synergistic antitumour effect in four pancreatic cancer cells (MIA PaCa‐2, CFPAC‐1, Panc‐1 and BxPC‐3). In addition, the GEM/NSC109555 combination significantly increased the level of intracellular reactive oxygen species (ROS), accompanied by induction of apoptotic cell death. Inhibition of ROS generation by N‐acetyl cysteine (NAC) significantly reversed the effect of GEM/NSC109555 in apoptosis and cytotoxicity. Furthermore, genetic knockdown of CHK2 by siRNA enhanced GEM‐induced apoptotic cell death. These findings suggest that inhibition of CHK2 would be a beneficial therapeutic approach for pancreatic cancer therapy in clinical treatment. 相似文献
108.
Soo-Jin Kim Yun-Hee Jang Moriyuki Hamada Jae-Hyung Ahn Hang-Yeon Weon Ken-ichiro Suzuki Kyung-Sook Whang Soon-Wo Kwon 《Journal of microbiology (Seoul, Korea)》2013,51(3):400-404
One bacterial strain 2RL3-2T was isolated from Chungkukjang, a traditional Korean fermented food made from soybeans, and determined to be a Gram-positive, aerobic, spore-forming rod. Growth of the novel strain was optimal at 30°C and pH 7.0. The 16S rRNA gene of strain 2RL3-2T showed the highest level of sequence similarity to Lysinibacillus sinduriensis BLB-1T (99.0%), Lysinibacillus massiliensis 4400831T (97.1%), Lysinibacillus xylanilyticus XDB9T (97.0%), and Lysinibacillus odysseyi 34hs-1T (96.8%). Phylogenetic analysis showed that strain 2RL3-2T formed a robust cluster with L. sinduriensis BLB-1T, L. massiliensis 4400831T, and L. odyssey 34hs-1T. The major fatty acids were anteiso-C15:0 (47.3%), iso-C16:0 (16.3%), and anteiso-C17:0 (11.3%), and the only menaquinone was MK-7. Diphosphatidylglycerol, phosphatidylglycerol, and phosphatidylethanolamine were the major polar lipids, along with an unknown phospholipid and two unknown lipids. The peptidoglycan type was A4α, with an interpeptide bridge of l-Lys-d-Asp. DNA-DNA hybridization values between strain 2RL3-2T and closely related Lysinibacillus species were below 43±4%. Therefore, based on phenotypic, chemotaxonomic, and phylogenetic characteristics, it was determined that strain 2RL3-2T represents a novel species of the genus Lysinibacillus, for which the name Lysinibacillus chungkukjangi sp. nov. is proposed. The type strain is 2RL3-2T (=KACC 16626T =NBRC 108948T). 相似文献
109.
Hong Min Jeong Kim Dae Yeon Choi Hong-Il Seo Yong Weon Kim Jin-Baek 《Molecular biology reports》2020,47(2):1129-1141
Molecular Biology Reports - F-box proteins play important roles in the regulation of various developmental processes in plants. Approximately 1796 F-box genes have been identified in the wheat... 相似文献
110.
Sung Han Kim Weon Seo Park Sun Ho Kim Boram Park Jungnam Joo Geon Kook Lee Jae Young Joung Ho Kyung Seo Jinsoo Chung Kang Hyun Lee 《PloS one》2016,11(3)
We aimed to identify tissue biomarkers that predict early biochemical recurrence (BCR) in patients with high-risk prostate cancer (PC), toward the goal of increasing the benefits of neoadjuvant hormonal therapy (NHT). In 2005–2012, prostatectomy specimens were collected from 134 PC patients who had received NHT and radical prostatectomy. The expression of 13 tissue biomarkers was assessed in the specimens via immunohistochemistry. Time to BCR and factors predictive of BCR were determined by using the Cox proportional hazards model. During the follow-up period (median, 57.5 months), 67 (50.0%) patients experienced BCR. Four (3.0%) patients were tumor-free in the final pathology assessment, and 101 (75.4%) had negative resection margins. Prostate stem cell antigen (PSCA) was the only significant prognostic tissue biomarker of BCR [hazard ratio (HR), 2.58; 95% confidence interval (CI), 1.06–6.27; p = 0.037] in a multivariable analysis adjusted by the clinicopathological variables that also significantly predicted BCR; these were seminal vesicle invasion (HR, 2.39; 95% CI, 1.32–4.34), initial prostate serum antigen level (HR 1.01; 95% CI, 1.001–1.020), prostate size (HR, 0.93; 95% CI, 0.90–0.97), and the Gleason score of preoperative biopsies (HR, 1.34; 95% CI, 1.01–1.79). We suggest that PSCA is a useful tissue marker for predicting BCR in patients with high risk PC receiving NHT and radical prostatectomy. 相似文献