Journal of the korean academy of Pediatric Dentistry
/
v.32
no.1
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pp.75-88
/
2005
Ameloblastoma is the most commonly occurring odontogenic tumor in oral cavity. Although most are benign epithelial neoplasm, they are generally considered to be locally aggressive and destructive, exhibiting a high rate of recurrence. The biological behavior of this neoplasm is a slowly growing, locally invasive tumor without metastasis, therefore malignant neoplasm, changed its histological appearance to carcinoma or showed distant metastasis, is only defined clinically. In this study, we identified the differentially expressed genes(DEGs) in stages under benign or malignant ameloblastoma compared with normal patient using ordered differential display(ODD) reverse transcription(RT)-PCR and $GeneFishing^{TM}$ technology. ODD RT-PCR is rather effective when the investigation of samples containing very small amounts of total RNA must be accomplished. ODD RT-PCR used the means of amplification with anchored T-primer and adaptor specific primer. bearing definite two bases at their 3' ends and so this method could display differential 3'-expressed sequence taqs(ESTs) patterns without using full-length cDNAs. Compared with standard differential display, ODD RT-PCR is more simple and have enough sensitivity to search for molecular markers by comparing gene expression profiles, However, this method required much effort and skill to perform. $GeneFishing^{TM}$ modified from DD-PCR is an improved method for detecting differentially expressed genes in two or more related samples. This two step RT-PCR method uses a constant reverse primer(anchor ACP-T) to prime the RT reaction and arbitrary primer pairs(annealing control primers, ACPs) during PCR. Because of high annealing specificity of ACPs than ODD RT-PCR, the application of $GeneFishing^{TM}$ to DEG discovery generates reproducible, authentic, and long(100bp to 2kb) PCR products that are detectable on agarose gels. Consequently, various DEGs observed differential expression levels on agarose gels were isolated from normal, benign, and malignant tissues using these methods. The expression patterns of the some isolated DEGs through ODD RT-PCR and $GeneFishing^{TM}$ were confirmed by Northern blot analysis and RT-PCR. The results showed that these identified DEGs were implicated in ameloblastoma neoplasm processes. Therefore, the identified DEGs will be further studied in order to be applied in candidate selection for marker as an early diagnosis during ameloblastoma neoplasm processes.
This study was carried out to decide ITS (internal transcribed spacer) sequence of some Korean native $Aster$ species and to resolve their relationship among Korean native $Aster$, including $Kalimeris$, $Gymnaster$, $Heteropappus$ genus separated from $Aster$ in a previously study based on the pappus length. We registered 11 ITS sequences of $Aster$ species including $A.$$glehni$ to GenBank and those sequences were used for the cluster analysis with $Kalimeris$ species. The size of ITS1 was varied from 248 to 256 bp, while ITS2 was varied from 220 to 222 bp. The G + C content of the ITS region ranged from 49.4 to 53.5%. Pairwise comparison results showed that the substitution rate of ITS1 and ITS2 region was 9% and 10%, respectively. $Kalimeris$ sensu strict substitution rate was lower than that of $Aster$ sensu strict species. The strict consensus parsimonious cluster analysis showed $A.$$tripolium$ is the first branching from the clade and the next is $A.$$scaber$. The $Kalimeris$ species except for the $A.$$hispidus$ were grouped into the same clade with high bootstrap value (91%) within $Aster$. $Gymnaster$ and $Heteropappus$ that has been classified by morphological characters were also grouped into broad sense $Aster$ clade. These results implied these three genera could be merged together into $Aster$ based on the ITS sequences.
This paper deals with the reproductive ecology e.g., number of the pre-spawning moults, morphological characteristics of the pre-spawning moult the common moult, daily ration druing a molting cycle mating behavior, structures of spermatozoa and spermatophore, structure of vas deferens, mechanisms of the oviposition and brooding into the egg-chambers, a suitable time for the artificial mating and fertilization, time sequence of the oviposition and brooding into egg-chambers from the copulation, responses to temperature and chlorinity on the egg development and hatching, effect of temperatures on duration of egg development, physical mechanism of the egg hatching, to make an attempt for the artificial spawning and brooding to establish a suitable system of the artificial seedling-production for the aquaculture. 1. Females molted commonly $8{\~}10$ times at an interval of $17{\~}18$ days at $28^{\circ}C,\;3.26\~4.35\%_{\circ}$ while the prespawning moltings were $4{\~}5$ times at an interval of $13{\~}14$ days. The suitable state for artificial copulation was within 14 hours elapsed from the prespawning moltings (most suitable state was within 8 hours). Males discharged a gelatinous spermatophore and placed it on the females sternum during copulation. Oviposition was seen $6{\~}17$ hours after copulation. External fertilization was considered to take place at oviposition. Fertilized eggs held in egg-chambers forming between pleopods were about $5000{\~}6000$ in females those sizes about 6.5 cm in body length. 2. Eggs immediately after oviposition were elliptic shape, measuring $0.58{\times}0.48$ mm up to hatching. Their sizes increased with egg development and finally reached $0.85{\times}0.54$ mm up to hatching. The relationship between the long axis of the egg(Y in U) and days elapsed(X) was expressed as Y= 5.60194 + 0.007358X. The eggs performed superficial cleavage and their cleavage furrows became visible at the 4-daughter-nucleus stage. The eggs showed normal development up to hatching at water temperature range of $22{\~}30^{\circ}C$ (optimum temperature : $26{\~}28^{\circ}C$) and at chlorinity range of $0.00\~6.64\%_{\circ}$ (optimun chlorinity : $2.21{\%}_{\circ}$). The relationship between incubation period (Y in days) and water temperature(X in $^{\circ}C$) could be expressed as Y= 50.803-1.3555X. The eggs hatched $12{\~}13$ days after oviposition at $28.0{\~}28.6^{\circ}C$ 3. The pre-spawning moltings were appreciably different in the morphologic structure from those of common moltings. Breeding setae and dresses were formed on the thoracic regions, abdominal epimerae and the bases of the first to fourth pleopods in order to prepare and support oviposition, transfering and supporting eggs in egg-chambers up to hatching. These supplementary breeding organs were observed only at reproductive seasons.
Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.
Kim Kye-Won;Ha Sun-Hwa;Cho Kang-Jin;Kim Eun-Ju;Lee Min-Kyung;Yu Jae-Ju;Kim Jong-Guk;Lee Shin-Woo
Journal of Plant Biotechnology
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v.32
no.3
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pp.167-173
/
2005
Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.
Two attenuated Cucumber mosaic virus (CMV) isolates, Paf-CMV and Rs2-CMV that had been selected from CMV isolates associated with satellite RNA (satRNA) were tested for cross-protection effect in pepper plants. The viruses selected as attenuated strains appeared to be identical serologically and physically to the challenge virus (Mf-CMV), but they were lower in the dilution end-point of infectivity of crude sap than Mf-CMV When symptoms were observed in several indicator plants after inoculation, Paf-CMV and Rs2-CMV were symptomless or showed mild mosaic symptoms while another satRNA isolate Ap-CMV developed severe mosaic symptoms on the leaves as Mf-CMV. The nucleotide sequences of the satRNAs were determined by sequencing full-length cDNA clones. Paf-, Rs2- and Ap-satRNAs were 386, 335, and 347 nucleotides long, respectively, The sequences were then compared with the other known Y-satRNA, revealing that nucleotide sequences of the satRNAs consisted of 5'- and 3'-terminal conserved regions. However variations occurred on the middle regions of the sequences, especially those related to symptom interference, showing significant differences between Paf-satRNA and other isolates. Infectious transcripts of Paf-satRNA and Rs2-satRNA induced mild mosaic symptoms in pepper plants when supported by genomic RNAs of Mf-CMV. Under greenhouse conditions, Paf-CMV and Rs2-CMV were tested for cross-protection effect in pepper and tobacco (Nicotiana tabacum cv, Xanthi nc) plants against Mf-CMV. No symptoms were developed on the plants vaccinated with Paf-CMV until 3 weeks after inoculation with the virulent strain; however another attenuated isolate, Rs2-CMV, showed less effectiveness in cross-protection. Depending on the concentration of the challenged virus, symptoms sometimes appeared later in the upper leaves. However, in plants challenged with low concentrations (below 0.2 mg/ml) of the challenge inoculum, symptoms caused by the virulent strain did not develop on the plants vaccinated with Paf-CMV. In the field experiments, the number of pepper plants with severe mosaic symptoms in the control plots was progressively increased after transplanting and reached approximately 50% after 50 days. On the other hand, the incidence of mosaic disease appeared very low on the plants that had received the protective inoculation with Paf-CMV.
A 14-year-old male patient with previous surgical repair of tetralogy of Fallot was admitted with hemodynamically significant ventricular tachycardia (VT). On preoperative electrophysiologic study (EPS), the morphology of documented VT was RBBB of vertical axis with 320 msec cycle length. The endocardial mapping during VT delineated the origin of VT at right ventricular outflow tract (RVOT), where the patch was attached. The clinical VT had a clockwise reentry circuit around the patch with the earliest activation at the same site seen during the preoperative EPS. The previously placed right ventricular outflow patch and fibrous tissue were removed. During a postoperative EPS, it was no longer possible to induce the VT. Ventricular tachycardia following repair of tetralogy of Fallot seen in this patient was caused by a macro-reentry around the right ventricular outflow patch. We were able to ablate the VT with the aid of a detailed mapping of its epicardial activation sequence.
Lee, H.J.;Lee, S.H.;Cho, Y.M.;Yoon, H.B.;Jeon, B. K.;Oh, S.J.;Kwon, M.S.;Yoon, D.H.
Journal of Animal Science and Technology
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v.46
no.6
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pp.947-956
/
2004
The primary role of lipoprotein lipase(LPL) is the hydrolysis of triglycerides(TG) from the core of triglyceride-rich lipoproteins such as chylomicrons and very low density lipoproteins in plasma. Fatty acids liberated by LPL on capillary endothelial surfaces are available for tissues as energy sources especially in muscles or for storage in the form of TG in adipose tissues. Therefore, as the candidate gene related to the carcass traits of the beef cattle, we have directly sequenced the exon 5${\sim}$exon 6 region in the bovine LPL gene for discovery of single nucleotide polymorphism(SNP) with 24 unrelated Hanwoo(Korean cattle). Novel eight sequence variants were detected: three loci on exon 5, three on intron 5 and two on exon 6. All SNPs identified were strongly linked each other, and one hundred twenty eight Hanwoo samples were genotyped one SNP on intron 5 using PCR-restriction fragment length polymorphism method by digestion with Hae III restriction enzyme. The allele frequency of the polymorphism was 0.76 and 0.24. The effects of this polymorphism on the breeding values of the carcass weight, loin muscle area, back fat thickness and marbling score were analyzed using least square methods of SAS GLM. The marbling score of BB genotype was significantly higher than those of AA and AB genotypes(P<0.05). This result indicates that this polymorphism may be associated with the variation of marbling score. Further study is warranted to investigate the phenotypic association in Hanwoo.
Recently, it was reported that certain polymorphisms in the growth hormone secretagogue receptor gene (GHSR) are associated with the growth of chickens. However, the correlation between GHSR polymorphisms and economic traits has not been investigated in Korean native chickens (KNCs). Therefore, the objective of this study was to confirm the suitability of the GHSR gene as a candidate for genomic selection and identify a genetic marker for KNCs. A total of 220 KNCs from six breeds raised at the National Institute of Animal Science were genotyped for the c.739+726 SNP in the GHSR gene using polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP), and the sequence for a subset of 30 birds was analyzed using direct sequencing. The association between the SNP genotypes and the economic traits of the KNCs was analyzed using the statistical package for the social science (SPSS) software program. The association analysis between the c.739+726T>C SNP and economic traits revealed that the SNP was significantly associated with body weight at 150 and 270 days (BW150 and BW270, respectively) in all KNCs (p<0.01), BW150 in KNC (Gary) (p<0.05), and egg production number in KNC (White, p<0.05). In addition, the SNPs discovered using direct sequencing (513A>G, 517A>T) had a significant effect on the body weight and egg production traits (p<0.05). In conclusion, these results might be useful as a basis for studies on the improvement of KNC breeds. Furthermore, these results suggest that the SNPs (c.739+726T>C, 513A>G, and 517A>T) located in the GHSR gene could be useful molecular genetic markers for KNCs.
Seo, Bo Yoon;Park, Chang Gyu;Jung, Jin Kyo;Cho, Jumrae;Lee, Gwan-Seok;Kim, Kwang-Ho
Korean journal of applied entomology
/
v.57
no.4
/
pp.393-399
/
2018
A loop-mediated isothermal amplification (LAMP) primer set (WBPH-65) was designed for the species-specific detection of white-backed planthopper (WBPH) Sogatella furcifera based on the full-length sequence of the internal transcribed spacer 2 (ITS2) (KC417469.1). The WBPH-65 primer set consists of six primers (total 165 bp), F3 (18 bp), B3 (18 bp), FIP (43 bp), BIP (40 bp), LF (21 bp), and LB (25 bp). After the LAMP reaction of three rice planthoppers, S. furcifera, Nilaparvata lugens, and Laodelphax striatellus, with the WBPH-65 primer set for 60 min at $65^{\circ}C$, the LAMP products were observed in the genomic DNA of S. furcifera only. According to the DNA amount of S. furcifera and incubation duration at $65^{\circ}C$, the difference of fluorescence relative to the negative control (0 ng) was clearly observed in a 40-min incubation with 10 and 100 ng or in case of 60-min incubation with 0.01, 0.1, 1, 10, and 100 ng. There was little difference in fluorescence between the negative control and all the other DNAs tested in 20- and 30-min incubations. On the other hand, the WBPH-65 primer set without LF and LB primers showed little amplification in the genomic DNAs of the three rice planthoppers, S. furcifera, N. lugens, and L. striatellus in a 60-min incubation. These results suggest that all six primers (F3, B3, FIP, BIP, LF, and BF) are necessary for the WBPH-65 primer set to detect S. furcifera within a 60-min incubation, and is able to discriminate S. furcifera from at least N. lugens and L. striatellus.
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