• 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.

• Journal of Microbiology
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• v.34 no.2
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• pp.151-157
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• 1996

By PCR amplification using the sequence of the previously cloned shiga-like toxin II DNA, a gene encoding it has been cloned from an isolate of healthy Korean native bovine feces, Escherichia coli KSC109. The nucleotide sequence s included tow open reading frames coding for 319 and 89 amino acids corresponding to A and B subunits, respectively. Comparison of the nucleotide and predicted amino acid sequences of newly cloned gene (slt-II) with those of others in the SLT-II family revealed completely identical homology with SLT-II cloned previously from bacteriophabe DNA of E. coli 933 derived from a patient with hemorrhagic colities. In addition, the sequence homology of SLT-II with SLT-II variant form bovine was more than 95% at both the nucleotide and protein levels. Overexpression of SLT-II recombinant gene by induction with IPTG using an E, coli hostvector, system was conducted and the correctly processed products with active mature form exhibited 1000-fold higher cytotoxycity for Vero cells than that form original strain.

• Journal of Plant Biology
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• v.39 no.4
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• pp.265-271
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• 1996

Full-length cDNA for RNA2 of cucumber mosaic virus strian Kor (Kor-CMV) was cloned downstream of synthetic T7 promoter by reverse transcriptase-polymerase chain reaction (RT-PCR). The clone could generate a full-length transcript corresponding to RNA1 in size when synthesized by T7 RNA polymerase. The complete nucleotide sequence has shown that the RNA2 is composed of 3,049 nucleotides and contains one functional open reading frame (ORF) of 2,574 nucleotides encoding 2a protein. The deduced translation product of the 2,574 nucleotides contains GDD motif which is a characteristic of RNA-dependent RNA polymerase (RdRp). The amino acid sequence analysis of the 2a protein has shown that the homology is found in decreasing order with O-CMV (98.8%), Y-CMV (98.7%), Fny-CMV (98.3%), KCMV (94.9%), Ix-CMV (91.9%), and Q-CMV (74.9%). Kor-CMV is suggested to belong to subgroup Ⅰ in the aspect of nucleotide sequence homology of RNA2.

• Mycobiology
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• v.28 no.4
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• pp.171-176
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• 2000

We have cloned a ${\alpha}-COP$ homolog, ancop, from Aspergillus nidulans by colony hybridization of chromosome specific library using ${\alpha}-COP$ homologous fragment as a probe. The probe DNA was amplified with degenerated primers designed by comparison of conserved region of the amino acid sequences of Saccharomyces cerevisiae ${\alpha}-COP$, Homo sapiens HEP-COP, and Drosophila melanogaster ${\alpha}-COP$. Full length cDNA clone was also amplified by RT-PCR. Comparison of genomic DNA sequence with cDNA sequence obtained by RT-PCR revealed 7 introns. Amino acid sequence similarity search of the anCop with other ${\alpha}-COPs$ gave an overall identity of 52% with S. cerevisiae, 47% with human and bovine, 45% with Drosophila and Arabidopsis. In upstream region from the transcription start site, a putative TATA and CAAT motif were also identified.

• Journal of Plant Biology
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• v.39 no.3
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• pp.197-202
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• 1996

A pollen-specific cDNA clone, LMP50, was isolated from the mature pollen cDNA library of the Easter lily. The LMP50 transcript was highly abundant in mture pollen grains but not detectable in other organs. The LMP50 cDNA clone contains 1383 nucleotides and two open reading frames. The first codes for a peptide of 15 amino acid residues. The role of this peptide is nuclear. The second encodes a protein containing 329 amino acid residues. This protein exhibited a significant homology to human tartrate-resistant acid phosphatase and porcine uteroferrin. Both of these enzymes have been suggested to play a role in iron transport. Therefore, LMP50 may act as an iron carrier protein in mature pollen grains.

• Journal of Life Science
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• v.9 no.4
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• pp.341-347
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• 1999

Insects use chitinolytic enzyme to digest chitin in the exoskelton during the molting process. We have isolated and sequenced a chitinase-encoding cDNA from the silkworm, Bombyx mandarina, compared its sequenced with genes encoding chitinolytic enzymes from other sources. The insert DNA in the clone is 2,675 nucleotides long with an open reading frame of 1,695 uncletides that encodes a protein of 565 amino acids with a molecuar weight of 63.4 kDa. The 3' -untranslated region of 889 nucleotides is AT-rich and contains two putative polyadenylation signals. The N-terminal sequence of the encoded protein contains numerous hydrophobic residues characteristic of a leader peptide. The amino acid alignment revealed that the endo-$\beta$-N-acetylglucosaminidase had 83% and 97% homology with M. sexta and B. mori, respectively. The deduced amino acid had two highly conserved region at the amino acid residues 97-111 and 139-148 that were related to the existing chitinase.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.118-120
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• 2002

Genetic materials including DNA plasmid are effective delivery vehicle to express interesting gene efficiently and safely not to generate replication competent virus. Moreover, it has advantages to design a better vector and to simplify manufacturing and storage condition. To understand a possible pathogenic mechanism by a flavivirus, West Nile virus (WNV), WNV genome sequence was aligned to other pathogenic viral genome. Interestingly, WNV capsid (Cp) amino acid sequence has some homology to HIV-l Vpr protein. These proteins induce apoptosis in human cell lines as well as in vivo and cell cycle arrest. Therefore, DNA plasmid carrying apoptosis-inducing and cell cycle arresting viral proteins including a HIV-1 Vpr and a WNV Cp protein can be useful for anti-cancer therapeutic applications. This WNV Cp protein is an early expressed protein which can be a reasonable target antigen (Ag) for vaccine design. Immunization of a DNA construct encoding WNV Cp protein induces a strong Ag-specific humoral and Th1-type immune responses in animal. Therefore, DNA plasmid encoding apoptotic viral proteins can be useful tool for therapeutic and prophylactic applications.

• Journal of Photoscience
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• v.4 no.3
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• pp.141-145
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• 1997

Polymerase chain reaction(PCR) was conducted with a pea cDNA library using two primers synthesized from homology analysis of amino acid sequences for animal and plant cytosolic FBPases. A PCR product with 650 bp long was cloned into pGEM-T vector and sequenced. The deduced amino acid sequence of the cDNA fragment was 98, 91, and 85% homologous with those of cytosolic FBPases from spinach, sugarbeet, and sugarcane, respectively. It was 51% homologous with amino acid sequence of FBPase from pea chloroplasts. Northern blot analysis was proceeded with the cDNA clone resulting that 1.2 kb transcript was highly expressed in light-grown pea leaves but almost not expressed in dark-grown etiolated pea seedlings. When peas grown in the light for 10 days were transferred to darkness, the transcript was gradually decreased with dark treatment, indicating that the expression of the enzyme was induced by continuous white light but suppressed by dark treatment. Pea cytosolic FBPase was highly expressed in leaves with trace amounts in stems. but almost not expressed in roots.

• Journal of Plant Biology
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• v.32 no.1
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• pp.33-40
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• 1989

Polysomal polyadenylated mRNAs which were purified from pea leaves were fractionated by sucrose grandient sedimentation. Fractions corresponding to the peak at 11.5S were found to contain mostly mRNA encoding the precursor polypeptide to the small subunit of ribulose bisphosphate carboxylase (rbcS) by in vitro translation in wheat germ extract. Double-stranded cDNA which was synthesized from the 11.5S mRNA was cloned into Hind III site of plasmid pBR 325. A cDNA clone, H24, was identified to code for rbcS. In vitro translation product of the hybridization-selected mRNA was molecular weight 20,000, presumably the precursor of rbcS. The nucleotide sequences of the H24 showed almost complete homology with the sequences encoding the transit peptide of the rbcS-3A gene which was reported by Fluhr et al.(1986).

• Journal of Sericultural and Entomological Science
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• v.41 no.3
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• pp.174-179
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• 1999

The genetic relationships among six Cordyceps spp. were investigated based on internal transcribed spacer sequences of ribosomal DNA .A portion of the these genes was amplified by PCR. Approximately 590 base pairs were successfully amplified, cloned, sequenced, compared. The nucleotide sequence of the six amplified fragments were aligned by the clustal W program. As a result, Cordyceps militaris shared 87, 96, 98, 90 and 97% sequences homology with Paecilomyces japonica, Paecilomyces sp. J300, Paeciomyces farinosa. Paecilomyces sp. J500 and Cordyceps sinensis, respectively. Paecilomyces japonica also shared 87, 88, 92 and 87% sequence similarity with Paecilomyces sp. J300, Paecilomyces farinosa, Paecilomyces sp. J500 and Cordyceps sinensis, repectively.