• 한국산업융합학회 논문집
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• 제14권1호
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• pp.9-14
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• 2011

The excess molar enthalpies $H_m^E$ at T=298.15 K and p=101.3 kPa of ternary system {1,2-dichloropropane (1,2-DCP) + 1,3-dioxolane+ 1,4-dioxane} were predicted by using the binary contribution model of $Radojkovi{\check{c}}$ with correlated sub-binary Redlich-Kister parameters. Excess partial molar enthalpies ${\bar{H}}_i^E$ were also calculated for the binary systems {1,2-dichloropropane + 1,3-dioxolane}, {1,2-dichloropropane + 1,4-dioxane} and {1,3-dioxolane + 1,4-dioxane} using adjustable parameters of Redlich-Kister equation. By extrapolation of excess partial molar enthalpies to infinite dilution, limiting excess partial molar enthalpies ${\bar{H}}_i^{E,{\infty}}$ of each component were also obtained. The ternary excess molar enthalpies excess partial molar enthalpies of these sub-binary systems have been calculated by using our previously reported results.

• 한국미생물·생명공학회지
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• 제51권1호
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• pp.26-31
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• 2023

A map gene encoding methionyl-specific aminopeptidase (MAP; EC 3.4.11.18) was cloned from Tetragenococcus halophilus CY54. Translated amino acid sequence of CY54 MAP showed high similarities with those from Enterococcus faecalis (83.8%) and Streptococcus salivarius (62.2%) but low similarities with MAPs from Lactobacillus and Lactococcus genera. The map gene was overexpressed in E. coli BL21(DE3) using pET26b(+),pET26b(+), and the recombinant MAP was purified by using an Ni-NTA column. The size of recombinant MAP was 29 kDa as determined by SDS-PAGE. The optimum pH and temperature of CY54 MAP were pH 5.0 and 60℃, respectively. The activity of CY54 MAP was most significantly increased by Co²⁺ ion (159%), and showed the highest activity at 12% NaCl. K_m and V_max were 0.64 ± 0.006 mM and 10.12 ± 0.014 U/mg protein, respectively when met-pNA was used as the substrate. This is the first report on a MAP from Tetragenococcus species.

• Asian-Australasian Journal of Animal Sciences
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• 제33권4호
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• pp.547-555
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• 2020

Objective: Apoptosis of ovarian granulosa cells (GCs) affects mammalian follicular development and fecundity. This study aimed to explore the regulatory relationship between microRNA-26a (miR-26a) and the 3β-hydroxysteroid-Δ24-reductase gene (DHCR24) gene in porcine follicular granular cells (pGCs), and to provide empirical data for the development of methods to improve the reproductive capacity of pigs. Methods: The pGCs were transfected with miR-26a mimic, miR-26a inhibitor and DHCR24-siRNA in vitro. The cell apoptosis rate of pGCs was detected by the flow cytometry. The secretion levels of estradiol (E2) and progesterone (P) in pGCs were detected by enzyme-linked immunosorbent assay. Double luciferase validation system was used to detect the binding sites between miR-26a and DHCR24 3'-UTR region. Qualitative real-time polymerase chain reaction and Western blotting were used to verify the DHCR24 mRNA and protein expression in pGCs, respectively, after transfecting with miR-26a mimic and miR-26a inhibitor. Results: Results showed that enhancement of miR-26a promoted apoptosis, and inhibited E2 and P secretion in pGCs. Meanwhile, inhibition of DHCR24 also upregulated the Caspase-3 expression, reduced the BCL-2 expression, promoted pGCs apoptosis, and inhibited E2 and P secretion in pGCs. There were the binding sites of miR-26a located within DHCR24 3'-UTR. Up-regulation of miR-26a inhibited DHCR24 mRNA and protein expression in pGCs. Conclusion: This study demonstrates that miR-26a can promote cell apoptosis and inhibit E2 and P secretion by inhibiting the expression of DHCR24 in pGCs.

• ETRI Journal
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• 제34권5호
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• pp.649-654
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• 2012

This paper presents a novel 16-quadrature-amplitude-modulation (QAM) E-band communication system. The system can deliver 10 Gbps through eight channels with a bandwidth of 5 GHz (71-76 GHz/81-86 GHz). Each channel occupies 390 MHz and delivers 1.25 Gbps using a 16-QAM. Thus, this system can achieve a bandwidth efficiency of 3.2 bit/s/Hz. To implement the system, a driver amplifier and an RF up-/down-conversion mixer are implemented using a $0.1{\mu}m$ gallium arsenide pseudomorphic high-electron-mobility transistor (GaAs pHEMT) process. A single-IF architecture is chosen for the RF receiver. In the digital modem, 24 square root raised cosine filters and four (255, 239) Reed-Solomon forward error correction codecs are used in parallel. The modem can compensate for a carrier-frequency offset of up to 50 ppm and a symbol rate offset of up to 1 ppm. Experiment results show that the system can achieve a bit error rate of $10^{-5}$ at a signal-to-noise ratio of about 21.5 dB.

• 한국미생물·생명공학회지
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• 제23권3호
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• pp.288-296
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• 1995

To investigate high-level expression of Serratia marcescens metalloprotease (SMP) in Escherichia coli and S. marcescens, we constructed various recombinant plasmids: pSP2, containing SMP gene and lac promoter; pKSP2, containing SMP gene and tac promoter; pTSP2, containing SMP gene, trc99a promoter, and lacI$^{q}$. The recombinant E. coli (pKSP2) strain expressed SMP to a high-level, about 36% of total cellular proteins but accumulated inactive SMP precursors intracellularly, which indicated that E. coli does not have activation and secretion system for SMP. To overproduce active SMP, we transformed S. marcescens with the recombinant plasmids by a modified CaCl$_{2}$ method. The recombinant S. marcescens ATCC27117 (pSP2) containing lac promoter for SMP transcription produced 530 U/ml of active SMP on LB broth, which is about 5.1 times of the SMP yield, 105 U/ml of a control strain, S. marcescens ATCC27117 (pUC19). However, S. marcescens ATCC27117 (pKSP2) containing tac promoter for SMP transcription did not grow healthy and hardly produced SMP. To overcome a harmful effect of the strong tac promoter, we constructed a regulatory plasmid pTSP2 containing a strong trc99a promoter and its repressor gene lacI$^{q}$. When S. marcescens ATCC27117 (pTSP2) was induced with 1.0 mM IPTG after 9 hr cultivation, 2,200 U/ml of SMP was obtained in LB broth, which is about 21 times of that of a control strain.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.65-75
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• 2003

Objectives: To investigate the role of TGF (Transforming growth factor-$\beta$) involved in the paracrinic communication during decidualization between UEC (uterine epithelial cells) and USC (uterine stromal cells), we have employed a co-culture system composed of human endometrial epithelial and stromal cells in defined hormonal conditions. Design: In the co-culture, endometrial epithelial cells cultured in the matrigel-coated cell culture insert are seeded on top of the endometrial stromal cells cultured within a collagen gel. The co-culture was maintained for 48 hours under the following hormonal conditions: progesterone dominant condition (100 nM P4 and 1 nM E2) or estrogen-dominant condition (100 nM E2 and 1 nM P4). 10 ng/ ml HGF and/or 10 ng/ml TGF-$\beta$1 are added. Methods: RT-PCR is utilized to detect mRNAs quantitatively. Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining are utilized to detect proteins in the tissue. Results: Prolactin mRNA is expressed in the co-cultured stromal cells under the progesterone dominant condition. TGF-$\beta$1 and its receptors are expressed in both the co-cultured epithelial and stromal cells irrespective of the steroid present, which is in contrast with no or negligible expression of TGF-$\beta$1 or its receptor in cells separately cultured. Both estrogen and progesterone significantly elevate the concentration of hepatocyte growth factor (HGF) in the conditioned medium of the co-culture with the value of 4, 325 pg/ml in E2-dominant and 2, 000 pg/ml in P4-dominant condition compare to 150 pg/ml in no hormone. In separately cultured stromal cells, administration of HGF induces the expression of TGF receptor 1 in both hormonal conditions, but induction of TGF receptor 2 is only manifest in the P4-dominant condition. Administration of TGF-$\beta$ and HGF directly induce the decidualization marker prolactin mRNA in separately cultured stromal cells. Conclusion: It is likely that steroid hormones induces prolactin mRNA indirectly by promoting the cell to cell communication between the stromal and the epithelial cells. TGF-$\beta$ and HGF are two possible paracrine mediators in the human endometrial decidualization.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권5호
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• pp.389-392
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• 2004

In order to produce a harmful protein more efficiently, this expression cassette, dubbed pCol-MICT, is directed by the colicin promoter, and was constructed by the insertion of a $rrnBT_1T_2$ fragment of pEXP7, and a MxelnteinCBD fragment of pTXB3, into pSH375. To test whether harmful proteins, including proteolytic enzymes, could be effectively produced by this cassette, the carboxypeptidase (CPase) Taq gene was inserted into the pCol-MICT cassette to yield pCol-CPase Taq-MICT. E coli W3l 10 tells harboring pCol-CPase Taq-MICT produced a large quantity of this enzyme, as much as 47.2 mg of purified from per liter of culture, when cultured in the presence of mitomycin C ($0.4{\mu}g/mL$). This indicates that the colicin promoter-controlled E, coli expression cassette was able to produce almost 8 times of protein than the conventional tar promoter-based system, and that this cassette may be useful in the Synthesis of other harmful proteins.

• 한국양식학회지
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• 제8권3호
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• pp.209-220
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• 1995

미꾸라지의 간으로부터 핵을 분리하여, 저농도 염추출 및 제한효소 처리로 핵기질(nuclear matrix)을 분리하였다. 분리된 핵기질을 Proteinase K로 분해한 후, phenol-chloroform 추출로 크기가 약 0.3kb-15kb의 분포를 나타내는 핵기질 부착 DNA (nuclear matrix attachment regions : MARs)를 얻었다. 효모 URA 3 유전자를 가진 2.13 kb Eco47 III 단편을 제한효소 Ssp I 으로 절단된 pUC19 플라즈미드 벡타에 결합시켜, ARS (autonomously replication sequence) 클로닝을 위한 pURY19 플라즈미드 벡타를 만들었다. 이 pURY19 벡타는 Saccharomyces cerevisiae내에서 독립적으로 복제할 수 없기 때문에, 물고기의 효율적인 발현 벡타 개발을 위해, 이 system을 이용하여, S. cerevisiae내에서 독립적으로 복제 가능한 미꾸라지의 ARS를 클로닝하고자 하였다. 분리 된 MARs를 pURY19 벡타에 결합시 킨 다음, E. coli $DH5\alpha$에 형질전환시켜 $pURY19N_{l-62}$를 얻었다. MAR Libraries $(pURY19N_{1-62})$를 각각 $Ura^-\;S.\;cerevisiae$에 형질전환시켜, S. cerevisiae내에서 독립적으로 복제 가능한 M. mizolepis 유래의 복제원점들 (ARSs)을 분리하여, Sanger's dideoxy-chain termination method로 염기서열을 분석하였다. 염기서열 분석결과 모든 clones들은 AT-rich하였으며, 특히 $pURY19N_6$에는 ARS concensus sequence, Topoisomerase II consensus, near A-box, 그리고 T-box들이 존재하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권5호
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• pp.726-732
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• 2014

The objective of this study was to investigate the effect of maintenance system as well as the effect of Se, Zn, and vitamin E supplementation of ram-lambs on the slaughter value and concentration of mineral elements in the loin muscle of lambs. The experiment was conducted on 72 Polish Merino ram-lambs divided into three groups: group C, indoor with no supplement, 19 lambs; S, indoor with supplement, 23 lambs; G, outdoor with no supplement, 30 lambs. From birth all the lambs were maintained indoor with their dams and then weaned at the age of 8 weeks. The rams from group C and S were placed in individual straw-bedded pens and fattened individually with concentrate mixture offered ad libitum until the age of 16 weeks. The lambs from group G were grazed every day from May to July (2 months). During the fattening period each lamb from the supplemented group S was administered per os 1 mL 0.1% $Na_2SeO_4$ (Se, 0.42 mg), 3 mL 10% $ZnSO_4$ (Zn, 68 mg), and 1 mL premix protect vitamin E (0.1 g ${\alpha}$-tocopherol, 5 mg lysine, 5 mg methionine) daily. A comparison of half carcasses across the groups has shown no difference between the control group and the one with supplements, while the weight of half carcasses in the grazing group was smaller in comparison with groups C and S (p<0.001). The meat content in the pelvic limb showed no differences across all groups under study. The pelvic limb of grazing lambs contained less fat compared to the control and supplemented groups (p<0.001). The concentrations of Se and Zn in the blood plasma of ram-lambs from the supplemented group were significantly higher than for the control and grazing lambs. Inorganic Se and Zn supplementation with vitamin E to the diet of lambs increased Se and Zn levels in loin muscle (p<0.001) to $0.46{\mu}g/g$ and $32.9{\mu}g/g$ in fresh tissue, respectively.

• Biomolecules & Therapeutics
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• 제11권3호
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• pp.157-162
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• 2003

The generation of secretory IgA antibodies(Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimHIctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was analyzed. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/CTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of CTXB. This study also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin/CTXA2B chimeric protein might be a potential antigen for oral immunization against UPEC.