• Journal of Plant Biology
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• v.35 no.3
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• pp.229-235
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• 1992

Differential expression of the three chlorophyll afb binding (cab) protein gene (cabl, cab2, and cab3) promoters of Arabidopsis thaliana was studied in tobacco plants transformed with cab-CAT (chloramphenicol acetyltransferase) translational fusions. CAT activity was measured to monitor the activities of the cab promoters. The activity of cabi promoter was higher than the other two in transformed tobacco leaves and also in calli and shoots derived from the leaves. Their activities were organ-specific and were the lowest in roots, medium in stems, and the highest in leaves. The relative activity of cabi promoter in stems comparing to it activity in leaves was, however, much higher than the values of cab2 and cab3. When the cab promoter activity was expressed as CAT activity per unit chlorophyll instead of CAT activity per unit protein, the relative cab] promoter activity (stem/leaf) became almost unity. This result suggests that cab2 and cab3 show photosynthetic organ-specificity but cabl does not. Similar result was obtained in the differentiation process of stems and leaves from shoots derived from the transgenic tobacco leaves.leaves.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1151-1160
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• 1999

This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

• Food Science and Biotechnology
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• v.17 no.2
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• pp.438-441
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• 2008

A novel lactobacilli replicon from plasmids of Lactobacillus reuteri KCTC 3678 was isolated. Eight L. reuteri strains from Korean Collection for Type Cultures (KCTC) and Korea Food Research Institute (KFRI) were screened for cryptic plasmids and most strains harbored 1 or 2 plasm ids. Particularly, L. reuteri KCTC 3678 contained 6 plasm ids which all were used for screening of lactobacilli replicon. EcoRI digests of the plasmid DNA prep from L. reuteri KCTC 3678 were ligated with pUC19 and the recombinant DNAs were serially named from pLR1 to pLR7. A cat (chloramphenicol acetyltransferase; $Cm^r$) gene originated from pC194 was introduced into pLR1-7, resulting in pLR1cat-pLR7cat, respectively. The recombinant plasmids were introduced into L. reuteri KCTC 3679, and only transformants harboring pLR5cat were obtained, indicating that the insert in pLR5 functioned as a lactobacilli replicon.

• Journal of Embryo Transfer
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• v.23 no.2
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• pp.127-131
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• 2008

Bone marrow (BM) cell harvesting is a crucial element in the isolation of mesenchymal stem cells (MSCs). A simple method for harvesting cat BM cells is described. The results show that a large number of BM cells can rapidly be harvested from the cat by this simple procedure. MSCs prepared by density-gradient method were spindle-shaped morphology with bipolar or polygonal cell bodies and strongly positive for CD9 and CD44 and negative for CD18 and CD45-like. They were capable of differentiation to adipocytic and osteocytic phenotypes when exposed to appropriate induction media. The advantages of this method are its rapidity, simplicity, low invasiveness, and low donor attrition and good outcome.

• Korean Journal of Ichthyology
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• v.9 no.1
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• pp.91-98
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• 1997

The transgene, pFV4CAT, containing CAT reporter gene regulated by carp $\beta$-actin promoter, was expressed in independent transgenic mud loach germ lines, determined by reverse transcriptase-PCR (RT-PCR) and enzyme-linked immunosorbant assay (ELISA). Expression of the transmitted transgene was found to be tissue-specific in F1 and F2 generations. Tissue specificity of the expression was dependent on each transgenic line with reproducible patterns. Liver and spleen did express the transgene more frequently than other tissues tested, and muscle and heart revealed the higher amount of CAT than other tissues, while testes showed the lowest expression level. The highest level of CAT expression in muscle from a transgenic F1 line was corresponding to 68-fold compared to the basal levels of controls.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.9
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• pp.1328-1334
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• 2006

This paper aimed to study the toxicity of arsanilic acid on rat primary hepatocytes in vitro by a modification of the perfusion method. The conditions included concentrations of 0, 1.085, 10.85, 108.5, 1,085 and 10,850 mg/kg arsanilic acid in RPMI 1,640 medium at rat hepatocytes plates respectively, each group had five repeats at $37^{\circ}C$ for 48 h. The rat primary hepatocytes survival ratio, DNA Ladder, activities of glutathione peroxidase (GSH-px), superoxide dismutase (SOD) and catalase (CAT) in hepatocytes, activity of SOD in the medium and the expression of gene bax in hepatocytes were measured at 12 h, 24 h and 48 h respectively. The results showed that arsanilic acid decreased the activities of GSH-px and SOD, and increased the activity of CAT in all dosages, and affected as positive DNA ladder. Although the SOD activities of both hepatocytes and medium in 1.085 mg/L arsanilic acid were significantly lower than the base line at 12 h, CAT activity in 10.85 mg/L arsanilic acid was significantly higher than the base line at 48 h, and all of the DNA ladders were positive, which means 1.085 mg/L arsanilic acid induced apoptosis at 24 h. The gene expression of bax was significantly upregulated in 1.085 mg/L arsanilic acid or higher for 24 h.The parameters in 1,085 mg/L and 10,850 mg/L arsanilic acid had more severe changes than the others at any time indicating that these levels of arsanilic acid were toxic hazards for hepatocyte survival. It was concluded that arsanilic acid induced a dosage- and time-dependent gene expression of bax, 1.085 mg/L arsanilic acid could be involved in rat liver cell apoptosis at 24 h. Arsanilic acid as additives in livestock feed could present potential toxic implications for farm animals.

• YAKHAK HOEJI
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• v.45 no.1
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• pp.108-115
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• 2001

We have recently reported the development of a high efficiency expression vector, pCK, which can drive a high level of gene expression in mouse skeletal muscle. In this study, we tested the therapeutic potential of pCK expressing human VEGF165, pCK-VEGF in the rabbit ischemic hind limb model. To determine the optimal dose of plasmid DNA, various concentrations of pCK-CAT were injected into the muscle of a rabbit hind limb and the levels of CAT activity were determined. It was found that the expression level of the exogenously added gene became stable between 250 and 1,000 $\mu$g. Based on this result, we tested whether intramuscular transfer of 500$\mu$g of pCK-VEGF could actually modulate collateral vessel development in a rabbit ischemic hind limb model. It was found that relative to the control group injected with the pCK lacking the VEGF sequence, single intramuscular doses (500$\mu$g) of pCK-VEGF produced statistically significant augmentation of collateral vessels as determined by the angiographic vessel count, maximal blood flow by Doppler flowmeter and the number of capillaries by histology. These results suggest that a single 500$\mu$g-delivery of pCK-VEGF is potent enough to induce sufficient angiogenic activity and achieve therapeutic benefit on this rabbit model.

• Animal cells and systems
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• v.2 no.1
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• pp.117-122
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• 1998

D-raf, a Drosophila homolog of the human c-raf-1, is known as a signal transducer in cell proliferation and differentiation. A previous study found that the D-raf gene expression is regulated by the DNA replication-related element (DRE)/DRE-binding factor (DREF) system. In this study, we found the sequences homologous to transcription factor C/EBP, MyoD, STAT and Myc recognition sites in the D-raf promoter. We have generated various base substitutional mutations in these recognition sites and subsequently examined their effects on D-raf promoter activity through transient CAT assays in Kc cells with reporter plasmids p5'-878DrafCAT carrying the mutations in these binding sites. Through gel mobility shift assay using nuclear extracts of Kc cells, we detected factors binding to these recognition sites. Our results show that transcription factor C/EBP, STAT and Myc binding sites in D-raf promoter region play a positive role in transcriptional regulation of the D-raf gene and the Myo D binding site plays a negative role.

• Journal of Veterinary Clinics
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• v.33 no.3
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• pp.145-150
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• 2016

We determined the prevalence of feline hemotropic mycoplasma species including 'Candidatus Mycoplasma haemominutum', Mycoplasma haemofelis, and 'Candidatus Mycoplasma turicensis' in naturally infected feral cats in Jeonju, Korea. Forty six feral cats were evaluated by PCR assay targeting the 16S rRNA gene sequence. Nine cats (19.6%) were positive for 'Candidatus Mycoplasma haemominutum', 2 cats (4.3%) were positive for 'Mycoplasm a haemofelis', and 1 cat (2.2%) was infected with both 'Candidatus Mycoplasma haemominutum' and Mycoplasma haemofelis. 'Candidatus Mycoplasma turicensis' was undetected. Partial 16S rRNA gene sequences of Mycoplasma haemofelis were closely (> 96%) related to those from other countries. The amplification of hemoplasma DNA in these samples confirmed the presence of 'Candidatus M. haemominutum' and M. haemofelis in Korea.

• Journal of Applied Biological Chemistry
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• v.43 no.1
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• pp.1-4
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• 2000

Chloramphenicol acetyl CoA transferase (CAT) and angiotensin- converting enzyme inhibitory peptide (ACEI) were fused to C-terminal region of rice ubiquitin to examine the level of transcripts or enzyme activities in yeast. When two chimeric genes under an inducible Gall promoter control were transformed into Saccharomyces cerevisaie, both CAT and ACE inhibitory activities were enhanced by three to four-fold as compared to those containing no ubiquitin gene. However, the levels of transcripts of ubiquitin fused and un fused genes were not significantly different each other. Therefore, it was suggested that the expression of foreign genes was post-transcriptionally enhanced by fusion of plant ubiquitin in heterologous organisms such as yeast.