• BMB Reports
• /
• v.38 no.4
• /
• pp.386-390
• /
• 2005

Based on the reported cDNA sequences of $BmK{\alpha}Txs$, the genes encoding toxin $BmK{\alpha}Tx11$ and $BmK{\alpha}Tx15$ were amplified by PCR from the Chinese scorpion Buthus martensii Karsch genomic DNA employing synthetic oligonucleotides. Sequences analysis of nucleotide showed that an intron about 500 bp length interrupts signal peptide coding regions of $BmK{\alpha}Tx11$ and $BmK{\alpha}Tx15$. Using cDNA sequence of $BmK{\alpha}Tx11$ as probe, southern hybridization of BmK genome total DNA was performed. The result indicates that $BmK{\alpha}Tx11$ is multicopy genes or belongs to multiple gene family with high homology genes. The similarity of $BmK{\alpha}$-toxin gene sequences and southern hybridization revealed the evolution trace of $BmK{\alpha}$-toxins: $BmK{\alpha}$-toxin genes evolve from a common progenitor, and the genes diversity is associated with a process of locus duplication and gene divergence.

• Asian-Australasian Journal of Animal Sciences
• /
• v.25 no.6
• /
• pp.758-763
• /
• 2012

As a member of a subclass of immunophilins, it is controversial that FKBP38 acts an upstream regulator of mTOR signaling pathway, which control the process of cell-growth, proliferation and differentiation. In order to explore the relationship between FKBP38 and mTOR in the Cashmere goat (Capra hircus) cells, a full-length cDNA was cloned (GenBank accession number JF714970) and expression pattern was analyzed. The cloned FKBP38 gene is 1,248 bp in length, containing an open reading frame (ORF) from nucleotide 13 to 1,248 which encodes 411 amino acids, and 12 nucleotides in front of the initiation codon. The full cDNA sequence shares 98% identity with cattle, 94% with horse and 90% with human. The putative amino acid sequence shows the higher homology which is 98%, 97% and 94%, correspondingly. The bioinformatics analysis showed that FKBP38 contained a FKBP_C domain, two TPR domains and a TM domain. Psite analysis suggested that the ORF encoding protein contained a leucine-zipper pattern and a Prenyl group binding site (CAAX box). Tissue-specific expression analysis was performed by semi-quantitative RT-PCR and showed that the FKBP38 expression was detected in all the tested tissues and the highest level of mRNA accumulation was detected in testis, suggesting that FKBP38 plays an important role in goat cells.

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

• Journal of Microbiology and Biotechnology
• /
• v.11 no.3
• /
• pp.508-514
• /
• 2001

A stuructural gene (ycl) encoding novel yeast cell wall hydrolase, YCL, was cloned from alkalophilic Bacillus alcalophilus subsp. YB380 by PCR, and transformed into E. coli JM83. Based on the N-terminal and internal amino acid sequences of the enzyme, primers were designed for PCr. The positive clone that harbors 1.8 kb of the yeast cell wall hydrolase gene was selected by the colony hybridization method with a PCR fragment as a probe. According to the computer analysis, this gene contained a 400-base-paired N-terminal domain of the enzyme. Based on nucletide homology of the cloned gene, a 850 bp fragment was amplified and the C-terminal domain of the enzyme was sequenced. With a combination of the two sequences, a full nucleotide sequence for YCL was obtained. This gene, ycl, consisted of 1,297 nucleotides with 27 nucleotides with 27 amino acids of signal sequence, 83 redundant amino acids of prosequence, and 265 amino acids of the mature protein. This gene was then cloned into the pJH27 shuttle vector and transformed into the Bacillus subtilis DB104 to express the enzyme. It was confirmed that the expressed cell wall hydrolase that was produced by Bacillus subtilis DB104 was the same as that of the donor strain, by Western blot using polyclonal antibody (IgY) prepared from White Leghorn hen. Purified yeast cell wall hydrolase and expressed recombinant protein showed a single band at the same position in the Western blot analysis.

• Korean Journal of Veterinary Service
• /
• v.33 no.1
• /
• pp.13-21
• /
• 2010

Chicken anemia virus (CAV) has been recognized as an immunosuppressive agent and plays role as an etiological agent of multifactorial diseases in chicken. In this study, we investigated distribution of CAV antibody by ELISA and the virus gene by PCR in poultry farms in Jeongeup, Jeonbuk province. In the test using ELISA kit, 41 (95.3%) of 43 flocks and 88.6% of the individual chickens were positive, respectively. By PCR, 90.9% of the broiler breeders and 75.0% of White-semi breeders were found positive, respectively. All hatchery was negative by PCR. Of the clinical cases from 49 poultry flocks, 87.5% of flocks and 54.7% for each samples were found positive by ELISA, respectively. By PCR test, 21 (42.9%) of 49 flocks were positive. Major clinical signs of the infected flocks were growth retardation, femoral subcutaneous bleeding, depression, limping, and continuing selection. The genetic analysis of separate N genes of CAV showed highly homologous each other. The nucleotide sequence of field isolates had homology ranged from 99.9% to 97.5% with Chinese strains, and 99.9% to 99.6% with Japanese strain. Phylogenetic analysis based on the N gene of CAV isolates showed the closely relation with Chinese strains. The results of this survey could be used as basic data for development of vaccine.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.2
• /
• pp.287-296
• /
• 2007

A Lactobacillus paraplantarum strain producing a bacteriocin was isolated from kimchi using the spot-on-the lawn method and named L. paraplantarum C7 [15]. The bacteriocin, paraplantaricin C7, was found to inhibit certain Lactobacillus strains, including L. plantarum, L. pentosus, and L. delbrueckii subsp. lactis. It also inhibited Enterococcus faecalis, yet did not inhibit most of the other LAB (lactic acid bacteria) tested. The maximum level of paraplantaricin C7 activity was observed under the culture conditions of $25^{\circ}C$ and a constant pH of 4.5. Paraplantaricin C7 retained 90% of its activity after 10 min of treatment at $100^{\circ}C$ and remained stable within a pH range of 2-8. Based on a culture supernatant, paraplantaricin C7 was purified by DEAE-Sephacel column chromatography and $C_{18}$ reverse-phase HPLC. SDS-PAGE and activity staining were then conducted using the purified paraplantaricin C7, and its molecular mass determined to be about 3,800 Da. The 28 N-terminal amino acids from the purified paraplantaricin C7 were determined, and the structural gene encoding paraplantaricin C7, ppnC7, was cloned by PCR using degenerate primers based on the N-terminal amino acid sequence. The nucleotide sequences for ppnC7 and other neighboring orfs exhibited a limited homology to the previously reported plantaricin operon genes. Paraplantaricin C7 is a novel type II bacteriocin containing a double glycine leader sequence.

• Korean Journal of Microbiology
• /
• v.40 no.1
• /
• pp.1-6
• /
• 2004

Tetracycline resistant bacterial strains were isolated from 10 batches of Kimchi among 50 batches collected in Taegu restrict. The MIC of tetracycline ranged between 25 and> 100 ㎖/l. Total genomic DNA preparation from all 10 tetracycline resistant lactic acid bacterial isolates were subjected to PCR amplification with class-specific primers for tet(M) and tet(O). In only one isolate, HJ9, tet(M) was detected. By Southern blotting and hybridization with a tet(M)-specific probe, the tet(M) gene of HJ9 isolate could be localized on a plasmid. The partial nucleotide sequence and deduced amino acid sequence of tet(M) of HJ9 showed 90-99% and 94-100% homology to those of Gram positive bacteria, respectively. With sequencing of 16S rRNA, HJ9 isolate from Kimchi was identified as Lactobacillus sakei. From these results, Kimchi can be considered potential vehicle for the spread of antibiotic-resistant lactic acid bacteria along the food chain to the consumer.

• Korean Journal of Plant Resources
• /
• v.17 no.3
• /
• pp.239-247
• /
• 2004

A cyclophilin 1 cDNA clone(GenBank accession no.CF924191) was isolated from the taproot of C. lanceolata and designed as C1CyP1. Determination of the nucleotide sequence of C1CyPl identified an open reading frame of 525bp, which shared high homologies with cyclophilins that were previously reported in other organisms. The C1CyP1 amino acid sequence possesses 7 amino acid residue stretch(KSGKPLH) that is characteristic of plant cytosolic dehydrins. Currently available amino acid residues of plant cyclophilins were compared to examine their phylogenetic relationship to C1CyP1. In the phylogenetic analysis, based on the aligned sequences, C1CyP1 showed high homology with arabidopsis ROC2 and rice CyP1. The transcript that corresponded to C1CyP1 was abundant in callus, but only basal level of transcript was detected in stem, leaf and root. For the study in the defense mechanism against various stresses, we report expression patterns of this gene by quantative RT-PCR.

• Journal of Veterinary Science
• /
• v.23 no.4
• /
• pp.57.1-57.9
• /
• 2022

Background: Classical swine fever virus (CSFV), the causative agent of classical swine fever (CFS), is a highly contagious disease that poses a serious threat to Chinese pig populations. Objectives: Many provinces of China, such as Shandong, Henan, Hebei, Heilongjiang, and Liaoning provinces, have reported epidemics of CSFV, while the references to the epidemic of CSFV in Yunnan province are rare. This study examined the epidemic characteristics of the CSFV in Yunnan province. Methods: In this study, 326 tissue samples were collected from different regions in Yunnan province from 2015 to 2021. A reverse transcription-polymerase chain reaction (RT-PCR), sequences analysis, and phylogenetic analysis were performed for the pathogenic detection and analysis of these 326 clinical specimens. Results: Approximately 3.37% (11/326) of specimens tested positive for the CSFV by RT-PCR, which is lower than that of other regions of China. Sequence analysis of the partial E2 sequences of eleven CSFV strains showed that they shared 89.0-100.0% nucleotide (nt) and 95.0-100.0% amino acid (aa) homology, respectively. Phylogenetic analysis showed that these novel isolates belonged to the subgenotypes 2.1c and 2.1d, with subgenotype 2.1c being predominant. Conclusions: The CSFV was sporadic in China's Yunnan province from 2015 to 2021. Both 2.1c and 2.1d subgenotypes were found in this region, but 2.1c was dominant.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.32 no.2
• /
• pp.357-369
• /
• 2005

Streptococci are Gram-positive, facultative anaerobes and have no catalase activities. Among mutans streptococci containing ${\alpha}-type$ hemolytic activity, S. mutans is a causative agent for dental caries. As well as acid production yielding the demineralization of tooth enamel, adherence and colonization of S. mutans to the teeth are also important for its virulence. These early colonization are accomplished by the bacterial fibrillar protein, Antigen I/II (Ag I/II) and glucosyltransferase (GTF). Therefore, Ag I/II and GTF are reasonable targets for the development of vaccine against S. mutans GS-5. The ag I/II and gtfD genes from S. mutans GS-5 were cloned and sequenced. Sequence analyses showed the nucleotides sequence of cloned genes had high homology to the sequences previously reported. The sequence alignment of 280 nucleotides between the cloned Ag I/II and the available sequence of the corresponding S. mutans GS-5 showed the perfect match. Comparing with the sequence of gtfD from S. mutans UA159, the corresponding nucleotide sequence of S. mutans GS-5 showed some mismatches and the mismatches introduced changes in four residues out of 105 amino acids, yielding four missense mutations.