• Korean Journal of Medicinal Crop Science
• /
• v.10 no.3
• /
• pp.194-199
• /
• 2002

Extracted genomic DNA from 16 accessions of Codonopsis lanceolata collected from South Korea and the Baekdoo Mt. areas of China were analyzed for their genetic relationships by RAPD. Twenty 10-mer-oligonucleotide primers having reproductive polymorphism were selected for the RAPD analysis. The size of amplified DNA was almost between 125 bp and 2.0 kbp. Sixteen collected Codonopsis lanceolata were analyzed with 20 primers which generated 73(49.3%) polymorphic bands among 148 PCR products. The mean number of polymorphic bands were 7.4 and varied $1{\sim}9$ per primer. It was, thus, demonstrated that RAPD was useful for detecting polymorphism in Codonopsis lanceolata. The range of 1-F value(genetic similarity) was from 0.682 to 0.959. These results indicate variable genetic similarities. By UPGMA (Unweighted Pair Group Method using an Arithmetic average) cluster analysis based on 1-F value, genetic distance among the 16 collected Codonopsis lanceolata was $0.133{\sim}0.400$. It was certainly classified into two groups between collected accessions from Korea and China, and the genetic distance was about 0.281. Both accessions collected from Korea and China showed miner differences, while the genetic relationships of Tonghua Xian and Liuhe Xian from China was farthest with other accessions collected.

• Korean Journal of Microbiology
• /
• v.28 no.2
• /
• pp.91-97
• /
• 1990

Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.1.15.1.1) was isolated from human liver cDNA library of $\lambda$gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage $\lambda P_{L}$ regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.

• Applied Biological Chemistry
• /
• v.41 no.1
• /
• pp.42-46
• /
• 1998

Polymerase chain reaction (PCR) and Southern hybridization analyses were carried out to identify clones of silk worm thorn (Cudraria tricuspidata) and Rose of sharon (Hibiscus syriacus) which look like one tree with two ar three, branches or two or three different trees. For PCR five different PCR primers $(17{\sim}24\;nucleotides)$ are derived from CaMV 35S promoter, nopaline synthase terminator and coding region of thylakoid membrane protein gene. In the case of silk worm thorn, about 500 bp of PCR product was produced from DNAs of one tree or branch in the presence of 35S primer alone. Southern hybridization analysis of genomic DNAs hybridized with $^{32}P$ labeled PCR product showed that the same size of DNA fragments were hybridized with different intensities. In addition, PCR analyses using 20 different primers of OPERON 10-mer kits showed that only OPA01 primer produced PCR products of different size. These results indicate that two different trees of silk worm thorn combined to one tree. In the case of the Rose of Sharon, the same size of PCR products were produced from three different samples but Southern hybridization with the above PCR product as a probe did not show any hybridized bands. PCR analyses in the presence of OPERON 10-mers showed OPA04 and OPA13 produced different products including same sizes of products. These, results indicate that three different trees of the Rose of Sharon seem to be derived from the tree.

• Geophysics and Geophysical Exploration
• /
• v.19 no.1
• /
• pp.37-44
• /
• 2016

$CO_2$-EOR (Carbon Dioxide-Enhanced Oil Recovery), one of the enhanced oil recovery methods, helps to not only enhance the production of oil, but also store carbon dioxide in underground. However, if micro fractures occur when during the injection of $CO_2$, it is difficult to make permanent storage of $CO_2$ in reservoir and can cause contamination of groundwater and soil. Therefore, in this study, we performed microseismic monitoring to investigate the occurrence of fractures during the $CO_2$ injection at the Meruap oil reservoir, Indonesia. To pick the first arrivals of microseismic events, Improved MER (Modified Energy Ratio) method was used. After picking the first arrivals, hodogram analysis was carried out by using the data recorded at three component geophones to calculate the back azimuth of events. Finally, locations of microseismic events were decided by using the results of first arrival picking and hodogram analysis. Estimated locations showed that all microseismic events were occurred at surface and any fracture did not occur around the reservoir. Moreover, by analyzing noise characteristic, we confirmed that almost of picked first arrivals were due to the repetitive mechanical noise.

• Journal of Veterinary Clinics
• /
• v.32 no.1
• /
• pp.28-35
• /
• 2015

Aim of this study is demonstrate the feasibility of Laparoscopic gastrostomy (LG) tube placement in dogs by comparing with percutaneous endoscopic gastrostomy (PEG) tube placement, based on operative time, complications and gastro-peritoneal adhesion evaluation. Eight intact male beagle dogs were used in this study. Tri-Funnel Replacement Gastrostomy tube (Bard Inc., USA) of 20 Fr was used for LG technique and PEG kit (Ponsky "Pull" PEG Kit$^{(R)}$, Bard Inc., USA) with soft silicone retention dome consisting of a 20 Fr gastrostomy tube was used. Feeding via gastrostomy tube was performed in two weeks, maintenance energy requirement (MER) divided into 3 separate feeding. LG and PEG were evaluated at intraoperative, postoperative and postmortem period. Mean operative time for the PEG group was significantly shorter when compared with the LG group (p < 0.05). Successful maintenance of gastrostomy tube was confirmed in all dogs. Gastric and peritoneal wall adhesions were formed successfully in each group. The mean adhesion length (AL) and width (AW) were significantly larger in LG group compared with in PEG group (p < 0.05). The mean adhesion distance (AD) was not significantly different between two groups (p = 0.182). Consequently, LG is an effective minimally invasive, safe and easy to perform technique for providing enteral nutritional support in dogs.

• Korean Journal of Medicinal Crop Science
• /
• v.9 no.3
• /
• pp.225-231
• /
• 2001

This studies were conducted to provide the basic information on safflower collections and to identify the variations which could be utilized in safflower breeding programs, The RAPDs was used to clarify the genetic relationships among safflower collections and to classify them into distint genetic groups. Among 30 of 10 mer primers in RAPD analysis, twenty were selected as the appropriate primers for identification of the genetic characters in safflower collections. Amplified PCR showed the highly reproducible bands at $3.0{\sim}0.2kb$. The number of bands amplified with the each primer showed the variations ranged from 2 to 11, with the average of 5.6. Total of 111 bands were identified among 20 selected primers used in PCR reaction and 84 bands (75.7%) showed polymorphism. Based on the similarity value of 0.14 in dendrogram derived from the cluster analysis using RAPD-PCR, the 30 safflower collections were classified into 11 groups. The two main groups, VII and VIII included 7 collections (23%) and 8 collections (27%), respectively. Most of the collections in group VII were the Korean collections (85%).

• Applied Biological Chemistry
• /
• v.39 no.2
• /
• pp.112-117
• /
• 1996

Arginine decarboxylase (ADC) is the first enzyme in one of the two pathways of diamine putrescine biosynthesis in plants. The genes encoding ADC have previously been cloned from Escherichia coli, oat and tomato genome. Two degenerate oligonucleotides (17-mer) corresponding to two conserved regions of ADC were used as primers in polymerase chain reaction of rice (Oryza sativa L.) genomic DNA, and an approximately 1.0 kbp fragment was obtained. This amplified PCR product showed an open reading frame which contains 1,022 bp of nucleotide sequences. This PCR product was cloned into pGEM-originated T vector and the short 500 bp PstI digested fragment was subcloned into pGEM-3zf(+/-) vectors to facilitate sequencing. The nucleotide sequence of this PCR product showed about 74% and 70% identity with the same regions of the oat and tomato ADC cDNA sequences, respectively. The predicted amino acid sequence exhibited 45% and 62% identity with oat and tomato ADC polypeptide fragments, respectively. The sequence similarities of 34%, 47% and 38% were previously reported in oat and E. coli, tomato and oat, and tomato and E. coli ADC amino acids, respectively. Therefore, similarities and identities between rice and oat or tomato are remarkably higher than those others of the previous reports. In the highly conserved regions in both the amino acid sequence and spacing regions among the sequences of these three, rice ADC open reading frame also has the exactly same regions with the striking similarity. RNA blot analysis showed that hnc is expressed as a transcript of approximately 2.5 kbP in the rice seedling leaf tissues.

• The Korean Journal of Zoology
• /
• v.39 no.4
• /
• pp.352-361
• /
• 1996

The c-myc proto-onco9ene, one of the immediately early genes, is involved in ceflular proliferation and differentiation, and its biologleal function is regulated hy dimerization with a heterodimeric partner, myn. In the present study, gene expression patterns of c-myc and myn during mouse preimplantation embryo development were examined using a semi-quantitative reverse transcription-poiymerase chain reaction (RT-PCR). Myn transcripts were rather constitutively expressed throughout embryonic stages with a slight increase only at biastocyst stage. in contrast, expression of c-myc transcripts wm developmental stage-'pedfic. The c-myc transcripts were detected at 1-cell stage, declined abruptly at 2-cell stage and then increased gradually at blastocyst stage. To examine the possible role of myn during preimpiantation mouse embryo development, two myn antisense oligonucleotides spanning the tail of zipper dognain (myn2; 20-mer) and the second helix domain (myn3; 20-mer) were microinjected into the fertilized 1-cellembryos. Microinjection of myn2 and myn3 resulted in developmental tion at morula/biastocyst transition stage, leading to the fiagentation of embryos. Talien together, these results suggest that c-myc and its heterodimeric partner, myn, are differentially expressed In a developmental stage-dependent manner, and myn may play an important role in mouse preimpiantation embryo development.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.21 no.1
• /
• pp.53-66
• /
• 1995

The gene for ${\gamma}$-casein, a milk protein, is a member of a family of casein gene which is expressed in mammary glands of the animal during the late gestation and lactation periods binder the influence of various hormones. In order to elucidate tile mechanisms b)'which hormones regulate the coordinate induction of milk protein genes, the mouse ${\gamma}$-casein gene was isolated and characterized. The ${\gamma}$-casein gene was screened from a mouse genomic library constructed in bacteriophage EMBL3 with the ${\gamma}$-casein CDNA used as probe and one clone was obtained. The ${\gamma}$-casein CDNA as probe was partially sequenced and contained ATG start codon and 5'-noncoding region. The cloned genomic DNA was digested with Sal I restriction enzyme, by which the insert DNA can be isolated from EMBL3 vector. Three DNA bands were observed and the size of DNAs was approximately 28kb, 14kb and 9Kb, respectively Accordingly the size of the insert DNA was calculated with approximately 23Kb. The result of Southern blot analysis, however, showed that the cloned genomic DNA was not hybridized with the synthetic oligonucleotides (40 mer) of cDNA 5'-end region, but it was hybridized with the y -casein CDNA. This means that tile cloned y -casein gene may not contain its promoter region. The ${\gamma}$ -casein genomic DNA containing the promoter region has been screening from mouse genomic library with oligonucleotides of CDNA 5'-end region as probe, and twenty-nine clones was obtained.

• Journal of Life Science
• /
• v.13 no.4
• /
• pp.541-550
• /
• 2003

E2 glycoprotein of hepatitis C virus (HCV) comprises a surface of viral particle together with E1 glycoprotein, and is thought to be involved in the attachment of HCV viral particle to receptor (s) on the permissible cells including hepatocytes, B cells, T cells, and monocytes. We constructed a phage library expressing cellular proteins of hepatocytes on the phage surface, which turned out to be 8.8${\times}$$10^5$ cfu of diversity and carried inserts in 95% of library. We screened both cDNA phage library and 12-mer peptide library to identify the cellular proteins binding to E2 protein. Some intracellular proteins including tensin and membrane band 4.1 which are involved in signal transduction of survival and cytoskeleton organization, were selected from cDNA phage library through several rounds of panning and screening. On the contrary, membrane proteins such as CCR7, CKR-L2, and insulin-like growth factor-1 receptor were identified through screening of peptide library. Phages expressing peptides corresponding to those membrane proteins were bound to E2 protein specifically as determined by neutralization of binding assay. Since it is well known that HCV can infect T cells as well as hepatocytes, we examined to see if E2 protein can bind to CCR7, a member of C-protein coupled receptor family expressed on T cells, using CCR7 transfected tells. Human CCR7 cDNA was cloned into pcDNA3.1(-) vector and transfected into human embryonic kidney cell, 293T, and expressed on the surface of the cell as shown by flow cytometer. Binding assay of E2 protein using CCR7 transfected cells indicated that E2 protein bound to CCR7 by dose-dependent mode, giving rise to the possibility that CCR7 might be a putative cellular receptor for HCV.