• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.1
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• pp.16-22
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• 2005

cDNA for oxidosqualene cyclase was cloned by a homology-based PCR method and sequenced from Centella asiatica. In a sequences analysis, the putative polypeptide of C. asiatica cycloartenol synthase (CaCYS) deduced from the 2,274 bp nucleotide sequence, consisted of 758 amino acids and had a molecular mass of 86.3 kD. The predicted amino acid sequence exhibited high homology to that of PNX (cycloartenol synthase) from Panax ginseng ($89\%$). Southern blot analysis suggests that CaCYS may be present in one copy of the C. asiatica genome. If methyl jasmonate (MJ) is applied exogenously to plants, not only triterpene saponins are accumulated in tissues, but also it produces effects such as growth inhibition and the promotion of ethylene production. In order to investigate the effect of MJ and thidiazuron (TDZ), a cytokinin that plays a role as an antisenescence agent in several plants, on the level of CaCYS mRNA, we performed northern blot analysis. When MJ is alone treated by adding to culture medium, CaCYS transcripts were inhibited. However, sustained levels of the expression of CaCYS, by adding TDZ to the medium despite MJ treatments, were demonstrated in C. asiatica leaves.

• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.285-292
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• 1993

C. sinensis total RMh was containing large amount of 185 rRNA but little 285 rRNA. The size of the double-stranded cDNA synthesized from poly $(A)^{+}$ mRNA was 0.4-4.2 kb long with tapering unto 9.5 kb. Degenerated oligonucleotides (as 2 sense and 3 antisense Primers) were designed on the conserved regions of the known tropomyosin amino acid sequences. From one out of the PCR amplifications using total CDNA and matrix of primers, a specific gene product, 580 bp in size, was produced. Upon Southern hybridization of the PCR products with Schistosomn mnnsoni tropomyosin (SMTM) CDNA, only one signal appeared at the band of 580 bp product. This 580 bp product was considered to encode C. sinensis tropomyosin (CSTM) and cloned in pGEM-3Zf(-) for DNA sequencing. CSTM cDNA was 575 bp containing one open reading frame of 191 predicted amino acids, which revealed 86.3% homology with SMTM and 51.1% with rrichostronsylur coeubnlormis tropomyosin. CSTM cDNA obtained will serve as a probe in the studies of molecular cloning of CSTM.

• Journal of fish pathology
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• v.7 no.2
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• pp.95-103
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• 1994

We have prepared cDNA libray of loach. M. mizolepis in order to isolate cDNA clone of growth hormone gene. Total RNA was isolated from pituitary of loach, and then mRNA was further purified from total RNA by oligo (dT)-coupled magnetic beads. The purified mRNA was used as substrates to prepare cDNA. The resulting cDNA was ligated into the EcoRV/Smal site of pBlueKS+. The ligation mixture have transformed E. coli JM109 strain with electroporator to obtain high yield of transformation efficiency. All the transformants was screened with DIG-labeled Tilapia growth hormone gene by high density colony hybridization. After isolating 10 putative colonies showing the positive signals, secondary colony hybridization and southern hybridization could confirm it as true clones. The nucleotide sequence of one candidate, pCGHI, was compared with 312 bp DNA fragment used as DNA probe and show 52% relative homology to Tilapia growth hormone gene.

• The Plant Pathology Journal
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• v.23 no.4
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• pp.272-280
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• 2007

Isolates of Cucumber mosaic virus (CMV) collected in Korea, were compared with their pathological features in tobacco and zucchini squash. Full-length cDNA clone of RNA3 was generated by using long-distance RT-PCR. Transcript RNA3 from the cDNA clone was inoculated onto host plants with transcripts RNA1 and RNA2 of Fny strain, generating RNA3-pseudorecombinant CMV. Timing and severity of systemic symptom was not significantly different among the pseudorecombinant CMVs in tobacco, compared with strains Fny-CMV and Pf-CMV. However, the pseudorecombinant CMVs induced two different systemic symptoms (mosaic vs. chlorotic spot) in zucchini squash. Based on symptom induction, the pseudorecombinant CMVs were categorized into two classes. The severity and timing of symptoms were correlated with viral RNA accumulations in systemic leaves of zucchini squash, suggesting that different kinetics of virus movement associated with CMV proteins are crucial for systemic infection and symptom development in zucchini squash. The analysis of movement proteins (MP) of CMV strains showed high sequence homology, but the differences of several amino acids were found in the C-terminal region between Class-I-CMV and Class-II-CMV. The analysis of coat proteins (CP) showed that the CMV isolates tested belonged to CMV subgroup I and the viruses shared overall 87-99% sequence identity in their genomes. Phylogenetic analysis of MP and CP suggested that biological properties of Korean CMV isolates have relationships associated with host species.

• The Korean Journal of Food And Nutrition
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• v.19 no.4
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• pp.496-501
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• 2006

This study was to isolate a bacterium producing a alkaline protease from mud flats of the west seaside of Korea and to investigate the biochemical analysis of the alkaline protease producing from the isolate. The isolate was named as Gelidibacter sp. HK-1 based on 16S rRNA sequence, Gram staining and the photograph of electron microsceope. Optimum temperature for growth and pretense production of the isolate was $25^{\circ}C$. Growth of the isolate was reached at stationary phase after 10hrs followed by inoculation. Maximum activity of protease produced from the isolate was shown after 14hrs. Optimum temperature and pH for the protease activity were $45^{\circ}C$ and pH 9, respectively. Molecular weight of the pretense was about 50KD and the partial amino acid sequence of the pretense was Ala-Try-Ala-Leu-Asn-Thr-Ser-Val-Thr-Glu-Thr-Phe-Ala-Lys. The partial amino acid sequences of the protease showed significant homology with a pretense produced from Streptomyces avermitilis.

• Journal of Microbiology and Biotechnology
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• v.24 no.8
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• pp.1073-1080
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• 2014

A total of 10 cellulase-producing bacteria were isolated from soil samples irrigated with paper and pulp mill effluents. The sequencing of 16S rRNA gene revealed that all isolates belonged to different species of genus Bacillus. Among the different isolates, B. subtilis IARI-SP-1 exhibited a high degree of ${\beta}$-1,4-endoglucanase (2.5 IU/ml), ${\beta}$-1,4-exoglucanase (0.8 IU/ml), and ${\beta}$-glucosidase (0.084 IU/ml) activity, followed by B. amyloliquefaciens IARI-SP-2. CMC was found to be the best carbon source for production of endo/exoglucanase and ${\beta}$-glucosidase. The ${\beta}$-1,4-endoglucanase gene was amplified from all isolates and their deduced amino acid sequences belonged to glycosyl hydrolase family 5. Among the domains of different isolates, the catalytic domains exhibited the highest homology of 93.7%, whereas the regions of signal, leader, linker, and carbohydrate-binding domain indicated low homology (73-74%). These variations in sequence homology are significant and could contribute to the structure and function of the enzyme.

• Korean Journal of Poultry Science
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• v.33 no.4
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• pp.249-254
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• 2006

This experiment was conducted to examine the effective DNA related with muscle growth of Korean native chicken. cDNA library was constructed with mRNA subtraction from Korean native chicken to Cornish. Total mRNA was purified from pectoralis muscle of adult chicken. Five clones were compared their DNA sequence and characteristics based on GenBank. Clone NDS-1 (618nt) was low homology (10%) with other species, but it is closely related with triosephosphate isomerase which is play an important role in glycolysis. Clone NDS-6 (651nt) is corresponding to glyceraldehyde-3-phosphate dehydrogenase. These two clones are encoding to enzymes in key role in glycolysis. However, other three clones (NDS-2, NDS-10, NDS-12) have low homology with other species about 5.0%. These clones were not similar with any other eukaryotics. Therefore, three clones (NDS-2, NDS-10, NDS-12) are high possibility of specific DNA for muscle growth in Korean native chicken.

• Research in Plant Disease
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• v.14 no.3
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• pp.223-225
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• 2008

This study was carried out to identify the Rice black-streaked dwarf virus that infected maize plants collected from Gochang-gun in Jeollabukdo in 2005. The genomic dsRNA from infected plants was extracted and the genome pattern was analyzed by polyacrylamide gel electrophoresis. Results of the electrophoresis revealed the already known to-segment genome and the difference of mobility was confirmed in isolates by collected areas. The RBSDV was identified from the result of RT-PCR using the template of extracted dsRNA and specific primer. The results of S10 cloned to pGEM-T vector and the conducted in sequence analysis consisted of 1,801nt and 559aa. This was of the same size as the RBSDV S10 identified in rice, and the change was confirmed in 18 base and displayed homology of 99%.

• Microbiology and Biotechnology Letters
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• v.22 no.2
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• pp.134-142
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• 1994

The neucleotide sequences of the xylA gene encoding $\beta $-xylosidase of Bacillus stearothermophilus and is its flanking regions were datermined. Three open reading frame(ORFs) were found, one of which(ORF1) appeared to code for the $\beta $-xylosidase. The 1830 base pair ORF1 encoded 609 amino acids starting from a TTG initiation codon. The molecular weight deduced from the nucleotide sequence(68 KD) was in agreement with that estimated by SDS-polyacrylamide gel electrophoresis of the purified enzyme(66 KD). The Shine-Dalgarno sequence(5'-AGGAGG-3') was found 11 bp upstream of the initiation codon. Further 15 bp upstream, there observed a potential transcription initiation signals. The putative -10 sequence(CATAAT) and -35 sequence(TTGTTA) coresponded closely to the consensus sequences for Bacillus subtilis RNA polymerase with major sigma factor. The guanine-plus-cytosine content of the coding region of the xylA gene was 56mol% while that of the third position of the codons was 63 mol%. Based on the comparison with the amino acid sequences of several other carbohydrate degrading enzymes, two conserved regions, possibly participating in the catalytic mechamism of $\beta $-xylosidase xylA, were identified in 278-298 and 329-350 regions of the translated xylA gene. The nucleotide sequence of the xylA was found to exhibit no homology to any other genes so far reproted.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2005.11a
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• pp.64-65
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• 2005

Members of the Pumilio are the RNA binding proteins acting as translational repressors and required for germ cell development and asymmetric division. We identified chicken Pum1 and Pum2 that are similar to mouse and human in highly conserved C-terminal RNA-binding domain and eight tandem repeats. The comparative sequence analysis of Pum1 and Pum2 from fly, chicken, mouse and human shows high degree of evolutionary conservation in the homology of the peptide sequence and the structure of PUM-HD (Pumilio homology domain) with similar spacing between adjacent Pum repeats. Also, structures of chicken Pum1 and Pum2 genes are almost identical to those of mouse and human. We revealed that the expression levels of Pum1 and Pum2 were the highest in hatched female gonad among various embryonic tissues, and Pum2 expressed highly in 12-day and hatched gonad by real-time RT-PCR. These results suggest that Pum1 and Pum2 might have an effect on the development of chicken gonad.