• 식물분류학회지
• /
• 제52권4호
• /
• pp.269-274
• /
• 2022

Dracocephalum rupestre Hance is a perennial herb distributed across China, Mongolia, and Korea. This study reports the first complete chloroplast genome sequence of D. rupestre. The plastome is 151,230 bp long and exhibits a typical quadripartite structure comprising a large single-copy region of 82,536 bp, a small single-copy region of 17,408 bp, and a pair of identical inverted repeat regions of 25,643 bp each. It contains 130 genes, comprising 85 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Phylogenetic analysis of D. rupestre and related species of Lamiaceae showed that the genus Dracocephalum is a monophyletic group, and D. rupestre is most closely related to D. psammophilum.

• 식물분류학회지
• /
• 제53권3호
• /
• pp.230-236
• /
• 2023

Rhododendron caucasicum Pall. is a shrub distributed in the mountainous areas of the Caucasus from northeastern Türkiye towards the Caspian Sea. This study reports the first complete chloroplast genome sequence of R. caucasicum. The plastome is 199,487 base pairs (bp) long and exhibits a typical quadripartite structure comprising a large single-copy region of 107,645 bp, a small single-copy region of 2,598 bp, and a pair of identical inverted repeat regions of 44,622 bp each. It contains 143 genes, comprising 93 protein-coding genes, 42 tRNA genes, and eight rRNA genes. The large chloroplast genome size is likely due to the expansion of inverted repeats. A phylogenetic analysis of chloroplast genomes with other Rhododendron species supports previously recognized infrageneric relationship.

• Journal of Crop Science and Biotechnology
• /
• 제11권4호
• /
• pp.237-242
• /
• 2008

A liguleless mutant, which showed complete loss of lamina joint region at the junction between leaf blade and leaf sheath, was isolated from a Ds insertional mutants derived from the source cultivar, Dongjin. This mutant could not affect other developmental patterns like phyllotaxis. Southern blot analysis, using GUS as a probe, revealed that the liguleless mutant contained three Ds copies transposed in the rice genome. Among the four genomic sequences flanking the Ds, one was mapped in the intergenic region (31661640 - 31661759), and the other two predicted a protein kinase domain (12098980 - 12098667) as an original insertion site within a starter line used for massive production of Ds insertional mutant lines. Another predicted and inserted in first exon of liguleless 1 protein (OsLG1) that was mapped in coding region (LOC_Os04g56170) of chromosome 4. RT-PCR revealed that the OsLG1 gene was not expressed liguleless mutants. Structure analysis of OsLG1 protein revealed that it predicted transcription factor with a highly conserved SBP domain consisting of 79 amino acids that overlapped a nuclear localization signal (NLS). RT-PCR revealed that OsLG1 is mainly expressed in vegetative organs.

• Toxicological Research
• /
• 제18권4호
• /
• pp.411-416
• /
• 2002

Apoptosis, or programmed cell death, is a genetically regulated pathway that is altered in many cancers. Overexpression of bcl-2 leads to resistance to apoptosis and promotes tumorigenesis. To determine the effect of bcl-2 antisense treatment in human lung cancer cell lines, a 20 mer full phosphorothioate oligonucleotide (ODN) targeted at the coding region of the bcl-2 mRNA was synthesized. Western blot analyses were used to examine bcl-2 protein level in five human non-small cell lung cancer (NSCLC) cell lines (NCI-H226, SK-MES-1 NCI-H358, NCI-H522 and NCI-Hl 299) and four human small cell lung cancer (SCLC) cell lines (NCI-H69, NCI-H4l7, HCC-2108 and SW2). Three out of five NSCLC (NCI-H226, SK-MES-1 and NCI-Hl 299) and all of SCLC cell lines expressed Bcl-2 protein. Treatment of these cell with antisense ODN for 48 hours reduced their viability and Bcl-2 protein level. As a conclusion, bcl-2 antisense treatment appears reduction of the Bcl-2 protein levels and cytotoxic effect including apoptosis in human lung cancer cell lines.

• Asian-Australasian Journal of Animal Sciences
• /
• 제20권6호
• /
• pp.821-826
• /
• 2007

The aim of the experiment was to detect polymorphisms in the keratin-associated protein 8.2 (KAP8.2) gene to determine associations between the genotype and fibre traits in Chinese Inner Mongolia cashmere goats. The fibre traits data investigated were cashmere fibre diameter, combed cashmere weight, cashmere fibre length and guard hair length. Five hundred and forty-two animals were used to detect polymorphisms in the complete coding sequence of the hircine KAP8.2 gene by means of PCR-SSCP. The results identified six genotypes, AA, BB, DD, AB, AD and BD, coded for by three different alleles A, B and D. Two SNPs in the coding region were confirmed by sequencing, which were A214G and T218C respectively. The relationships between the genotypes and cashmere fibre diameter, combed cashmere weight, cashmere fibre length and guard hair length were analyzed. There were significant differences (p<0.01) between the associations of the different genotypes with cashmere fibre diameter, cashmere weight and hair length. Cashmere length was the only trait that was not associated with the genotypes. The genotype AA (0.73) was found to be predominant in Inner Mongolia cashmere goats and the animals with this genotype had the thinnest cashmere fibre diameter compared with the other genotypes. These results suggested that polymorphisms in the hircine KAP8.2 gene may be a potential molecular marker for cashmere fibre diameter in cashmere goats.

• Journal of Microbiology and Biotechnology
• /
• 제8권4호
• /
• pp.369-377
• /
• 1998

Rotavirus is a major cause of severe gastroenteritis in young children and animals throughout the world. The VP7 of rotavirus is thought to induce the synthesis of neutralizing antibodies and to be responsible for determining viral serotypes. The cDNA coding for the VP7 capsid protein of human rotavirus, obtained from Korean patients (HRV-Y14), was cloned and its nucleotide sequence was determined. Comparative analysis of the nucleotide sequences between VP7 of Y14 and that of other foreign isolates showed $92.7~95.2\%$ homology to G1 serotypes (RV-4, KU, K8, WA), $74.2\%$ homolgy to G2 serotype HU-5, $76.4\%$ homology to G3 serotype SA-11, and $77.6\%$ homology to G4 serotype A01321. These data suggest that HRV-Y14 can be classified as a G1 serotype. cDNA coding for VP7 of HRV-YI4 was subcloned into the baculovirus vector and the VP7 glycoprotein was expressed in insect cells. The expressed proteins in Sf9 cell extract and tissue culture fluid were separated on SDS-PAGE, and Western blot analysis with monoclonal antibody raised against the synthetic peptide containing 21 amino acids within the VP7 conserved region was performed. The molecular weight of recombinant VP7 was estimated to be 36 kDa which is about the same size as the native VP7. Addition of tunicamycin in the culture media caused a reduction of the molecular weight of the recombinant VP7 indicating that the expressed protein was glycosylated.

• Journal of Microbiology and Biotechnology
• /
• 제24권3호
• /
• pp.337-345
• /
• 2014

Pichia pastoris is one of the most widely used expression systems for the secretory expression of recombinant proteins. The secretory expression in P. pastoris usually makes use of the prepro $MAT{\alpha}$ sequence from Saccharomyces cerevisiae, which has a dibasic amino acid cleavage site at the end of the signal sequence. This is efficiently processed by Kex2 protease, resulting in the secretion of high levels of proteins to the medium. However, the proteins that are having the internal accessible dibasic amino acids such as KR and RR in the coding region cannot be expressed using this signal sequence, as the protein will be fragmented. We have identified a new signal sequence of 18 amino acids from a P. pastoris protein that can secrete proteins to the medium efficiently. The PMT1-gene-inactivated P. pastoris strain secretes a ~30 kDa protein into the extracellular medium. We have identified this protein by determining its N-terminal amino acid sequence. The protein secreted has four DDDK concatameric internal repeats. This protein was not secreted in the wild-type P. pastoris under normal culture conditions. We show that the 18-amino-acid signal peptide at the N-terminal of this protein is useful for secretion of heterologous proteins in Pichia.

• IMMUNE NETWORK
• /
• 제2권3호
• /
• pp.175-181
• /
• 2002

Background: S100A6 is a calcium-binding protein overexpressed in several tumor cell lines including melanoma with high metastatic activity and involved in various cellular processes such as cell division and differentiation. To detect S100A6 protein in patient' samples (ex, blood or tissue), it is essential to produce a monoclonal antibody specific to the protein. Methods: First, cDNA coding for ORF region of human S100A6 gene was amplified and cloned into the expression vector for GST fusion protein. We have produced recombinant S100A6 protein and subsequently, monoclonal antibodies to the protein. The specificity of anti-S100A6 monoclonal antibody was confirmed using recombinant S100A recombinant proteins of other S100A family (GST-S100A1, GST-S100A2 and GST-S100A4) and the cell lysates of several human cell lines. Also, to identify the specific recognition site of the monoclonal antibody, we have performed the immunoblot analysis with serially deleted S100A6 recombinant proteins. Results: GST-S100A6 recombinant protein was induced and purified. And then S100A6 protein excluding GST protein was obtained and monoclonal antibody to the protein was produced. Monoclonal antibody (K02C12-1; patent number, 330311) has no cross-reaction to several other S100 family proteins. It appears that anti-S100A6 monoclonal antibody reacts with the region containing the amino acid sequence from 46 to 61 of S100A6 protein. Conclusion: These data suggest that anti-S100A6 monoclonal antibody produced can be very useful in development of diagnostic system for S100A6 protein.

• 한국식물병리학회지
• /
• 제14권3호
• /
• pp.240-246
• /
• 1998

A cDNA encoding desiccation-related protein was isolated from a flower bud cDNA library of Chinese cabbage (C-DH) and its nucleotide sequence was characterized. It contains 679 bp nucleotides with 501 bp open reading frame. The amino acid sequence of the putative protein showed the highest amino acid sequence homology (79 % identity) to dehydrin protein in Gossypium hirsutum. Also, the C-DH shares 48-52% amino acid sequence identity with the other typical dehydrin proteins in plant cells. When the amino acid sequence of their proteins were aligned, several peptide motifs were well conserved, of which function has to be solved. Particularly the C-DH contains 15 additional amino acids at its N-terminus. Genomic Southern blot analysis using the coding region of C-DH showed that the C-DH consists of a single copy gene in Chinese cabbage genome. The C-DH mRNA, whose transcript size is 0.7 kb, was expressed with a tissue-specific manner. It was highly expressed in seed, flower buds and low expression as detected in root, stem or leaf tissues of Chinese cabbage. And the transcript level of C-DH was significantly induced by the treatment of plant hormone, abscisic acid and water-deficit conditions.

• BMB Reports
• /
• 제31권2호
• /
• pp.130-134
• /
• 1998

An aldolase gene has been cloned from Methanococcus jannaschii. The coding region of the gene has been expressed in E. coli using a pET system to a level of 30% of total cellular proteins. The protein was purified to more than 95 % homogeneity by heat treatment and ion exchange chromatography. The protein performed an aldol condensation reaction with glyceraldehyde as substrate and dihydroxyacetone phosphate as a carboxyl donor. The protein was determined to be a type II aldolase which requires the $Zn^{2+}$ ion as a metal cofactor. This enzyme has a broad range of optimum pH (7-9) and temperature ($50-80^{\circ}C$). It shows strong stability against heat, chemical denaturants, as well as a high percentage' of organic solvents. The half-life of this enzyme at $85^{\circ}C$ is more than 24 h and it maintains more than 90% of aldolase activity in the presence of 6 M urea, 50% acetonitrile, or 15% isopropyl alcohol.