• International Journal of Industrial Entomology and Biomaterials
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• v.3 no.1
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• pp.57-62
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• 2001

A cDNA encoding the defensin homologue of a novel family member was isolated from the cDNA library of the firefly,Pyrocoelia rufa. Sequence analysis of the cDNA encoding the defensin homologue of P. rufa resulted that the 165 bp cDHA has an open reading frame of 55 amino acid residues. The deduced amino acid sequences of the defensin homologue gene from P. rufa showed identity to known mammalian defensins. Also 6 cystein residues in the P. rufa defensin homologue gene were conserved in the same position as those of known mammalian defensins. The result suggested that P. rufa defensin homologue is a novel member of the insect defensin family. Southern blot analysis suggests that there may be a single copy number of the P.rufa defensin homologue gene and their fat body-specific expression pattern at the transcriptional level was confirmed by Northern blot analysis.

• BMB Reports
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• v.29 no.2
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• pp.146-150
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• 1996

A 2.1 kb cDNA clone for rat transketolase was isolated from rat liver ${\lambda}gt11$ cDNA library and its sequence was determined. The predicted rat transketolase (655 amino acids with $M_r$ 71,186) is highly similar (92%) to that of the human enzyme except that it contains an extra 32 amino acids at its N-terminus. Although it is less similar (<27%) to transketolases from non-mammalian species, the functional motifs such as the catalytic sites and thiamine binding domain are well conserved in the rat enzyme. Southern blot analysis of genomic DNA verified that transketolase appears to be derived from a single gene. Immunoblot and Northern blot analyses suggested that hepatic transketolase was activated pretranslationally by a 2.1-fold while little change was observed in brain enzyme, indicating a tissue-specific pretranslational activation during postnatal development.

• Plant Biotechnology Reports
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• v.4 no.3
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• pp.193-199
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• 2010

An olive-green mutant was generated in Synechocystis sp. strain PCC 6803 by inactivation of the sll0396 gene. Whole-cell absorption spectra of the mutant revealed the missing of phycocyanin peak. An investigation of the low-temperature fluorescence emission spectra revealed that the $sll0396{\Omega}$ mutant has a reduced amount of phycocyanin. Western blot analysis showed that the mutant contained less phycocyanin ${\beta}$- and ${\alpha}$-subunits and lacked the 30- and 32-kDa linker polypeptides, and northern blot analysis revealed that the transcription of the 1.4-kb cpcBA gene encoding the phycocyanin ${\beta}$- and ${\alpha}$-subunits was lower in the mutant. The Sll0396 protein has a DNA-binding motif and shares homology with known response regulators. Our results indicate that Sll0396 plays a regulatory role in the transcription of the phycocyanin genes during phycobilisome synthesis.

• Journal of The Korean Society of Grassland and Forage Science
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• v.20 no.2
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• pp.97-102
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• 2000

To increase thennotolerance of forage crops, transgenic rice plants as a model for transformation of monocots were generated. A cDNA encoding the chloroplast-localized small heat shock protein (small HSP) of rice, Oshsp21, was introduced into rice plants via Agrobacterium-mediated gene transfer system. Calli induced from scutella were co-cultivated with a A. tumefaciens strain EHAlOl canying a plasmid, pIGhsp21. A large number of transgenic plants were regenerated on a medium containing hygromycin. Integration of Oshsp2l gene was confirmed by PCR and Southern blot analyses with genomic DNA. Northern blot and immunoblot analyses revealed that the Oshsp21 gene was constitutively expressed and accumulated as mature protein in transgenic plants. Effects of constitutive expression of the OshspZl on thermotolerance were first probed with the chlorophyll fluorescence. Results indicate that inactivation of electron transport reactions in photosystem I1 (PSII), were mitigated by constitutive expression of the Oshsp21. These results suggest that the chloroplast small HSP plays an important role in protecting photosynthetic machinery during heat stress. (Key words : Thermotolerance, Rice, Transgenic, cDNA)

• Journal of Applied Biological Chemistry
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• v.44 no.1
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• pp.6-11
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• 2001

An ${\omega}-3$ fatty acid desaturase gene which is involved in de novo synthesis of -Iinolenate was isolated from cDNA library of Perilla frutescens. A cDNA library was constructed with mRNA extracted from perilla seeds of 12 DAF. The cDNA clone consisting of 1317-bp open reading frame encoding 438 amino acids with a relative MW of 50kDa, was isolated and showed 65-83% similarities to other known genes. This cDNA is deduced to encode a plastidal ${\omega}-3$ fatty acid desaturase based on the fact that it has higher homology to plastidal ones than to microsomal ones and its N-terminal sequence shares several characteristics of transit peptides of chloroplast proteins. Southern blot analysis of genomic DNA indicated that more than one gene or alleles for ${\omega}-3$ fatty acid desaturase are present in the genome of perilla. Northern blot analysis showed that the ${\omega}-3$ fatty acid desaturase gene is mainly revealed in early developing seeds and has different expression patterns depending on tissue types compared to the microsomal ones.

• Microbiology and Biotechnology Letters
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• v.25 no.5
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• pp.483-489
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• 1997

Sulforhodamine B (SRB) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay, RNA dot blot and Northern hybridization analysis were performed to screen microorganisms for the production of anticancer agent. Among microorganisms tested, strain YP-1 was selected for its cytotoxicity and ability to reduce the level of c-myc RNA. Strain YP-1 was identified as Streptomyces endus. The anticancer material produced by Streptomyces endus YP-1 was sequentially purified by solvent extraction, silica gel column chromatograpby, preparative TLC and preparative HPLC. The cancer material identified as azalomycin B by the instrumental analyses such as $^{1}$H-NMR, $^{13}$C-NMR, Mass, IR and UV absorption. It was colorless amorphous powder and its molecular weight was 1025.278. Azalomycin B, produced by Streptomyces endus YP-1, showed anticancer activity against several human cancer cell lines and reduction of c-Myc protein level in Colo320 DM cells which was determined by Western blot analysis.

• International Journal of Industrial Entomology and Biomaterials
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• v.17 no.2
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• pp.223-228
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• 2008

A novel beetle antimicrobial protein from stimulated Copris tripartitus and the corresponding gene were isolated in parallel through differential display-PCR and expression in Escherichia coli. To find cDNA clones responsible for bacteria resistance, the suppression subtractive hybridization and GeneFishing differentially expressed genes system were employed in the dung beetle, Copris tripartitus immunized with lipopolysaccaride. One cDNA clone from eight subtracted clones was selected through dot blot analysis and confirmed by northern blot analysis. The 516-bp, selected cDNA clone was determined by 5' and 3' rapid amplication of cDNA ends and cloned into the GST fusion expression vector pGEX-4T-1 for expression of the protein. The expressed protein was predicted 14.7 kDa and inhibited the growth of gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa. These results implied that the expressed protein is related to immune defense mechanism against microorganism.

• BMB Reports
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• v.30 no.4
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• pp.285-291
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• 1997

It is becoming increasingly evident that significant changes in gene expression occur during the course of neuronal differentiation. Thus, it should be possible to gain information about the biochemical events by identifying differentially expressed genes in neuronal differentiation The PC12 cell line is a useful model system to investigate the molecular mechanism underlying neuronal differentiation and has been used extensively for the study of the molecular events that underlie the biological actions of nerve growth factor (NGF). In this study, we report an application of the recently described mRNA differential display method to analyze differential gene expression during neuronal differentiation. Using this technique, we have identified several cDNA tags expressed differentially during neuronal differentiation. Interestingly, one of these clones was cytochrome c oxidase subunit I (COX I) gene. The differential expression of COX I gene was confirmed by Northern blot analysis as well as RT-PCR. Southern blot analysis of the genomic DNA of PC12 cells revealed that COX I is a single gene. Induction of the oxidative enzyme might reflect the energy requirement in neuronal differentiation.

• Journal of Plant Biotechnology
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• v.36 no.1
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• pp.45-52
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• 2009

In order to understand the molecular interactions that occur between rice and the rice blast fungus during infection, we previously identified a number of rice blast fungal elicitor-responsive genes from rice (Oryza sativa cv. Milyang 117). Here, we report the cloning and characterization of the rice fungal elicitor-inducible gene Oshin1 (GenBank Accession Number AF039532). Sequence analysis revealed that the Oshin1 cDNA is 1067 bp long and contains an open reading frame encoding 205 amino acid residues. The Oshin1 gene shows considerable sequence similarity to the tobacco hin1 and hin2 genes. The predicted Oshin1 protein has a cysteine-rich domain at the N-terminus and is rich in leucine, serine, and alanine residues. Southern blot analysis suggests that Oshin1 gene is a member of a small gene family in the rice genome. To examine the expression of Oshin1, Northern blot analysis was conducted. Expression of the Oshin1 transcript is rapidly induced in suspension-cultured rice cells treated with fungal elicitor, salicylic acid or hydrogen peroxide. In addition, Oshin1 transcript levels are rapidly increased by treatment with $Ca^{2+}$/A23187. The expression of Oshin1 was also elevated in 3-week old leaf tissues upon ethephon application or fungal elicitor treatment. Our results suggest that the Oshin1 gene is involved in plant defense responses to environmental stresses.

• Journal of Plant Biotechnology
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• v.35 no.1
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• pp.41-45
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• 2008

Laccase (EC 1.10.3.2) is a small group of enzymes that catalyze the oxidation of a broad range of phenolic compounds including hazardous and recalcitrant pollutants in the environment. This study attempted to develop an efficient system for production of a recombinant laccase by chloroplast genetic transformation of tobacco. Chloroplast transformation vector was constructed and introduced into the tobacco chloroplast genome using particle bombardment. Chloroplast-transformed plants were subsequently regenerated. PCR and southern blot analyses confirmed stable integration of the laccase gene into the chloroplast genome. Northern blot analysis revealed that mRNA of the laccase gene was highly expressed in chloroplast-transformed plants.