We have previously produced transgenic (TG) mice expressing the human lactoferrin (hLF), interleukin-10 (hIL-10), and thrombopoietin (hTPO) proteins in the milk. In this study, we examined whether simple crossbreeding between two kids of a single transgenic mouse can produce double transgenics co-expressing two human proteins.. The hLF male, and the hIL-10 male were crossbred with the hIL-10 and hTPO females, and the hTPO female, respectively. PCR analysis for genotyping showed 32%, 23% and 24% double transgenic rates for hLF/hIL-10, hLF/hTPO, and hIL-10/hTPO transgenes, respectively. We analyzed the expression levels of the human proteins from double transgenic mice and compared those with their single transgenic siblings. All double transgenic co-expressed two human proteins at comparable levels to singles', unless hTPO was not co-expressed: for hLF, 1.1 mg/ml in hLF/hIL-10, whereas 0.5 mg/ml in hLF/hTPO; for hIL-10, 4.1 mg/ml in hIL-10/hLF, whereas 1.4 mg/ml in hIL-10/hTPO. Ihe downregulation of hTPO to half level of singles' was observed in double transgenic mice. The possible reason why hTPO co-expressed might lead to down-regulation of another human protein was discussed. These results suggested that double transgenic generated by crossbreeding between two singles' could be useful system for bioreactor.

Acute lymphoblastic leukemia (ALL), a predominantly pediatric disease involving uncontrolled proliferation of white blood cells within the bone marrow, is strongly associated with chromosomal translocations. The human chromosomal translocation t(12;21)(p12;q22) is the most frequently encountered chromosomal rearrangement in pediatric B-lineage ALL, and results in fusion of the TEL and RUNX1 genes. The resulting TEL/RUNX1 fusion protein is generally thought to be a transcriptional repressor that interferes with RUNX1-mediated transactivation. We used a luciferase assay system to investigate the effects of TEL/RUNX1 and PEBP2β on RUNX1-mediated transactivation. TEL/RUNX1 blocked the synergistic transactivation achieved by PEBP2β and RUNX1, and coimmunoprecipitation and immunofluorescence analyses showed that PEBP2β interacted with TEL/RUNX1 and was sequestered in the cytoplasm. Theses results suggest that TEL/RUNX1 inhibits RUNX1-mediated transactivtion via cytoplasmic sequestration of coactivator(s) such as PEBP2β.

In this study 300 weaned pigs (Landrace × Yorkshire × Duroc, 23±3 d of age, 5.56± 1.21 kg initial body weight) were used to study the effect of fungal (Aspergillus oryzae, FSP-A) and fungal + bacteria (Aspergillus oryzae + Bacillus subtilis, FSP-B) fermented soya proteins on their blood hematology, enzymes and immune cell populations. Pigs were allotted to 5 treatments, each comprising of 4 pens with 15 pigs. Basal diets consisted of 15% soyabean meal (Control diet) while for treatment diets SBM was replaced with 3 and 6% of each FSP-A and FSP-B, respectively. The experimental diets were fed from 0 to 14 day after weaning and then a common commercial diet was fed from 15 to 35 day. Blood was collected on 14 and 35 day of experiment and analyzed for hematology, aspartate transaminase (AST), alanine transaminase (ALT) and immune cell populations. At d 14, lower RBC count, Hb and HCT values and higher AST values were noted in pigs fed FSP-A diets when compared with Control and FSP-B fed pigs.Also at d 14 pigs fed 6% FSP-A had lower NE (P<0.05) when compared with those fed 6% FSP-B. The level of FSP influenced the RDW on d 14 and MCHC, MO and MPV on d 35. In addition on d 35, pigs fed 3% FSP-A had lesser NE than those fed 6% FSP -A and Control diet, while pigs fed 6% FSP-B had the highest number of MO compared to other treatments. But there were no differences in the plasma AST and ALT values on d 35. Thus it may be concluded that the FSP either by fungal or fungal + bacterial sources had an influence on the blood hematological status and the populations of immune cells.

The human ELAV(embryonic lethal abnormal vision)-like protein HuR stabilizes a certain group of cellular mRNAs that contain AU- ri ch elements in theil‘ 3’• untranslated region, Dysregulation of mRNA s tability may be relevant in tumor biology and may lead to abnormal expression of several proteins in malignant tumors, The aim of this study is to identify the differentially expressed proteins according to the functi onal activi ty of HuR Methods : We used stabl e expression of small interfering RNA(siRNA) of HuR gene to inhibit the expression of HuR in human ora l car cinoma cell lines‘ KB cell line and YD10B cell line‘ and compared the proteomic changes between s iRNA-treated and cont rol cell line using two-dimensional gel electrophoresis , Flow cytometry caliber scan(FACS) was employed to investigate the effects of HuR on cell apoptosis and proliferation , Results: Seventeen differentially expressed proteins between the two cell lines were identified by electrospray ioruzati on quadrupole time-of- fl ight mass spectrometry(ESI-Q-TOF MS) and database searching, Among them there are eleven proteins which are significant ly up- regulated in siRNA- treated cell line, which include heat shock protein 10(influencing nucl eocytoplas rnic transpo 다， cell dedifferentiation , and inhibition of apoptosis) , keratin 19(basal cell differ ent iat ion) ‘ and nucleoside diphosphate kinase B(G protein activator), etc Enolase 1 and Ml of pyruvate kinase a re the representatives of signifi cantly down-regulated proteins in siRNA-treated cell 11l1e Conclusion : Our data suggest that HuR participate in mRNAs stability of proteins that have the counter effects in the carcinogenesis of oral ca ncer , And the functiona l proteomics a re needed to elucidate the detailed interactions between HuR and t hese molec ules

Transforming growth factor-β (TGF-β) has been shown to have a positive effect on in vitro fertilization (IVF) and has been reported to stimulate meiosis at follicular level in variety of species. The study was designed to determine the expression patterns of TGF-β1, TGF-β receptors type Ⅰ, Ⅱ and Smads gene in bovine oocytes and embryos. TGF-β1 and their receptors were observed in the unfertilized oocytes. TGF-β1 and type Ⅱ receptor were not expressed at the blastocyst stage, however, only type I receptor was exclusively observed at the same stage. The blastocyst stage, in particular, showed high levels of mRNA expression patterns containing a TGF-β type Ⅰ receptor. The mRNA expression pattern of Smad 2 at all stages of embryonic development was similar in all respect with TGF-β1 type I receptor. On the contrary, Smad 3 and 4 were expressed with high and low level mRNA at the blastocyst stage. In conclusion, it is suggested that TGF-β signaling may be regarded as an important entity during the preimplantation embryo development.

탁수가 어류의 조직과 혈장 단백질에 미치는 영향을 조사하기 위하여 탁수역인 임하호와 비탁수역인 안동호에 서식하는 치리 (Hemiculter eigenmanni)를 공시재료로 사용하였다. 탁수역에 서식하는 치리의 아가미에서는 이차새변의 만곡, 곤봉화, 부종 및 상피세포 박리 현상 등이 나타났고, 신장에서는 사구체의 크기가 다소 작아지는 수축현상과 조직의 괴사 및 공포현상이 관찰되었다. 또한 간세포의 수가 적어졌으며 크기가 불규칙한 형태로 나타났다. 주사전

This study was conducted to examine the effect of IRES controlled reporter gene on screening and production of recombinant human erythropoietin (EPO) proteins from cultured CHO cells. The cDNA was cloned for EPO from human liver cDNA. Using site-directed mutagenesis, we generated recombinant human EPO (rhEPO) with two additional N-glycosylations (Novel erythropoiesis-stimulating protein: NESP). Wild type hEPO and NESP were cloned into expression vectors with GFP reporter gene under regulatory control of CMV promoter and IRES so that the vectors could express both rhEPO and GFP. The expression vectors were transfected to cultured CHO-K1 cells. Under microscopy, expression of GFP was visible. Using supernatant of the culture, ELISA assay, immunocytochemistry and in vitro assay using EPO dependant cell line were performed to estimate biological activity to compare the production characteristics (secretion levels, etc.) between rhEPO and NESP. The activity of NESP protein, obtained by mutagenesis, was described and compared with its rhEPO counterpart produced under same conditions. Although NESP had less secretion level in CHO cell line, the biological activity of NESP was greater than that of rhEPO. These results are consistent with previous researches. We also demonstrated that rhEPO and GFP proteins expressed simultaneously from transfected CHO cell line. Therefore we conclude that use of GFP reporter gene under IRES control could be used to screen and produce rhEPO in cultured CHO cells.

실리카 입자를 기공 형성제로 사용하여 다공성 키토산 및 키틴 막을 제조하였다. 다공성 막의 제조는 다음의 3단계 절차로서 수행되었다: (1) 키토산 용액에 실리카 입자를 첨가시켜 필름을 형성시킨 후, (2) 이 필름을 알카리 용액에 침지시켜 실리카 입자를 제거하여 다공성의 키토산 막을 제조하였으며, (3) 다공성 키토산 막을 acetic anhydride를 사용하여 아세틸화시킴으로서 다공성 키틴 막을 제조하였다. 물리적 강도가 우수하고, 적절한 순수 투과량을 갖는 다공성 키토산 막과 키틴 막의 최적 제막조건이 제시되었다. 단백질 친화성을 부여하기 위해 다공성 키토산 막에 반응성 염료인 Cibacron Blue 3GA를 고정화시켰으며, BSA 단백질 및 lysozyme 효소의 흡착실험을 수행하여 친화 키토산 막 및 키틴 막의 단백질 결합용량을 측정하였다. 친화 키토산 막의 BSA 단백질 결합용량은 약 22 mg/mL이었으며, 친화 키틴 막의 lysozyme 효소 결합용량은 약 26 mg/mL로서 이는 키토산 또는 키틴을 기반으로 하여 제조된 hydrogel bead의 단백질 결합용량보다 수~수십 배 큰 값으로서, 향후 막여과 크로마토그래피용 친화 막으로의 효과적인 활용이 기대된다.