• The Korean Journal of Food And Nutrition
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• v.9 no.4
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• pp.452-458
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• 1996

A PPIase gene of Bacillus stearothermophilus was screened from a genomic library by plaque hybridization using the A-1 primer as a probe. A PPIase positive plaque contained a 3.0kb insert of the chromosomal DNA. A 3.0kb fragment was subcloned into pUC18, resulting pPI1-40. A DNA fragment encoding the N-terminal portion of the PPIase in pPi-40 was amplified by polymerase chain reaction(PCR) method using the A-1 and B-2 primers. The amplified fragment was cloned into the Sma I site of pUC18 and recombinant plasmid was designated as pSN-18. The nucleotide sequence of 167bp fragment was determined. The deduced amino acid sequence of PPIase was completely matched with the determined N-terminal amino acid sequence of PPIase B. stearothermophilus. The translated protein sequence of PPIase B. stearothermophilus was compared with sequence from periplasmic PPIase from Escherichina coil ; homogies of 16 and 58%, respectively, were found. The clond PPIase gene was over-expressed in E. coil cell using pUC19 as an expression vector. The enzyme was partially purified by heat treatment and colum chromatochraphy on DEAE-Sepharose CL-6B. The molecular weight of the enzyme was dermined to be about 18.0 kDal by SDS-PAGE.

• The Journal of the Korea institute of electronic communication sciences
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• v.13 no.6
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• pp.1397-1404
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• 2018

Due to the technical difficulties and high cost for obtaining 3-dimensional structure data, sequence-based approaches in proteins have not been widely acknowledged. A motif can be defined as any segments in protein or gene sequences. With this simplicity, motifs have been actively and widely used in various areas. However, the motif itself has not been studied comprehensively. The value of this study can be categorized in three fields in order to analyze the human proteins using artificial intelligence method: (1) Based on our best knowledge, this research is the first comprehensive motif analysis by analyzing motifs with all human proteins in Protein Data Bank (PDB) associated with the database of Enzyme Commission (EC) number and Structural Classification of Proteins (SCOP). (2) We deeply analyze the motif in three different categories: pattern, statistical, and functional analysis of clusters. (3) At the last and most importantly, we proposed random motif frequency(RMF) matric that can efficiently distinct the characteristics of proteins by identifying interface residues from non-interface residues and clustering protein functions based on big data while varying the size of random motif.

• Journal of Animal Science and Technology
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• v.51 no.1
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• pp.1-8
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• 2009

We generated 57,598 expressed sequence tags (ESTs) from 3 cDNA libraries of Hanwooo (Korean Cattle), fat, loin, liver. Liver, intermuscular fat and longissimus dorsi tissues were obtained from a 24-month-old Hanwoo steer immediately after slaughter. cDNA library was constructed according to the oligocapped method. The EST data were clustered and assembled into unique sequences, 4,759 contigs and 7,587 singletons. To carry out functional analysis, Gene Ontology annotation and identification of significant leaf nodes were performed that were detected by searching significant p-values from $2^{nd}$ level GO terms to leaf nodes using Bonferroni correction. We found that 13, 26 and 8 significant leaf nodes are unique in the transcripts according to 3 GO categories, molecular function, biological process and cellular component. Also digital gene expression profiling using the Audic's test was performed and tissue specific genes were detected in the above 3 libraries.

• The Korean Journal of Pesticide Science
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• v.11 no.4
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• pp.320-330
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• 2007

Mitochondrial 16S DNA sequences of Bemisia tabaci which were collected on rose greenhouse of Iwol and Jinchen in Chungbuk and red pepper field of Miryang, Gyeongnam, were analyzed. The mtCOI PCR product of B. tabaci collected on red pepper field of Miryang were digested with EcoT14I (Sty I) into two fragments 555bp and 311bp, while the PCR product of B. tabaci collected on rose greenhouse of Iwol were digested with Sty I into two fragments of 560bp and 306bp. As a result, B. tabaci collected on red pepper reveal biotype Q and those on rose greenhouse was biotype B. These was difference between two biotypes in insecticide susceptibility, and the biotype B was more susceptible than biotype Q. As a result of foliar systemic test, root-uptake systemic test and residual effect, the biotype B was more susceptible. In case of inhibition effect on enzyme activities of fenitrothion (organophosphorous) and fenothiocarb (carbamate), those of biotype Q was higher than those of biotype B. These results indicate that biotype Q was more resistant than biotype B against 12 insecticides.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.117-123
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• 2003

For screening of autoregulator receptor gene from Saccharopolyspora erythraea, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHI site of pUC19 and transformed into the E. coli DH5$\alpha$. The isolated plasmid from transformant contained the fragment of 120 bp, which was detected on 2% gel after BamHI treatment. The insert, 120 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridization using Saccha. erythraea chromosomal DNA were performed with the insert as probe. The plasmid (pEsg) having 3.2 kbp SacI DNA fragment from Saccha. erythraea is obtained. The 3.2 kbp SacI DNA fragment was sequenced by the dye terminator sequencing. The nucleotide sequence data was analyzed with GENETYX-WIN (ver 3.2) computer program and DNA database. frame analyses of the nucleotide sequence revealed a gene encoding autoregulator receptor protein which is a region including KpnI and SalI sites on 3.2 kbp SacI DNA fragment. The autoregulator receptor protein consisting of 205 amino acid was named EsgR by author. In comparison with known autoregulator receptor proteins, homology of EsgR showed above 30%.

• Food Science of Animal Resources
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• v.27 no.3
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• pp.351-356
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• 2007

The objective of this study was to develop a rapid and reliable method for the sex determination of beef using the PCR(polymerase chain reaction) technique. We have used two bovine sex determining genes, SRY and ZFY, on the Y-chromosome to identify the sex of Hanwoo and Holstein beet. We attempted to amplify 1,348 bp and 979 bp fragments from male and female genomic DNA corresponding to the SRY and ZFY genes, respectively, using male specific primers. The amplified PCR products were separated by electrophoresis in a 1.5% agarose gel to detect a male specific DNA band. When DNA from male beef was amplified with primers specific for the SRY gene, a DNA band of 1,348 bp was present in all of the male samples, but absent from all of the female samples. Also, when DNA from male beef was amplified with primers specific for the ZFY gene, a DNA band of 979 bp was observed in all of the male samples, but absent from all female samples. In conclusion, the bovine SRY and ZFY genes are typically found only in male beef. For the practical application of this method for the sexing of commercial beef at the processing and marketing stages after slaughter. a total of 350 beef samples collected randomly from local markets were analyzed for sex determination. The proportions of male and female samples were 252 (72%) and 98 (28%), respectively. Therefore. the SRY and ZFY genes. which are specific for the Y-chromosome, may be useful sex-diagnostic DNA markers to distinguish male meat from female meat.

• Microbiology and Biotechnology Letters
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• v.42 no.4
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• pp.407-412
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• 2014

A number of Saccharomycopsis fibuligera strains that can hydrolyse and utilize starch as a carbon source were isolated from nuruk, a traditional Korean starter for rice wine fermentation, and their specific growth rates on starch-containing medium were compared to choose the prominent strain. S. fibuligera strain MBY1320 showed a higher growth rate at $42^{\circ}C$ than that of strain S. fibuligera KCTC7806, indicating that S. fibuligera MBY1320 has more thermo-tolerant machinery for starch hydrolysis and utilization than KCTC7806. Although the activity of ${\alpha}$-amylase at $30^{\circ}C$ was significantly lower for S. fibuligera MBY1320 than KCTC7806 (3,812.5 U vs. 14,878.5 U), S. fibuligera MBY1320 showed a much higher glucoamylase activity at $42^{\circ}C$ than S. fibuligera KCTC7806 (5,048.9 U vs. 13,152.3 U). Thus, a new S. fibuligera strain, with a higher starch-hydrolysing activity at elevated temperatures than that of other types of strain, this study reports.

• Journal of Animal Science and Technology
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• v.45 no.6
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• pp.911-916
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• 2003

The mitochondrial DNA(mtDNA) D-loop region was amplified from Duroc(Sus scrofa) by polymerase chain reaction(PCR). The oligonucleotide primer used to amplify the Sus scrofa mtDNA D-loop region was designed using tRNA-Pro and tRNA-Phe sequence in mtDNA regions highly conserved in many other animal species. There were 1,145 base pairs(bp) in the D-loop region. The middle of the region contained 10 tandem repeat of an 10-bp Sus scrofa-specific sequence, TACACGTGCG. We designed primers for PCR-mediated single stranded conformation polymorphism(SSCP) analysis that amplified a 345 bp fragment, which contained the most variable region according to our sequencing data. SSCP analysis of denatured amplification products was carried out by polyacrylamide(8%) gel electrophoresis followed by ethidium bromide staining. The SSCP analysis identified two band patterns(A and B) and comparision of these two nucleotide sequences identified 21 base substitutions. These results show that SSCP analysis of the D-loop region is useful for detecting the genetic polymorphism.

• Journal of Life Science
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• v.21 no.10
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• pp.1434-1442
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• 2011

A genetic variation within 29 strains of F. velutipes was analyzed by internal transcribed spacer (ITS) sequence analysis and random amplified polymorphic DNA (RAPD). Seven hundred and twenty base pairs were sequenced during the analysis of the ITS region, but no significant variation was observed among the 29 strains of F. velutipes. Sixteen out of 40 random primers amplified polymorphic RAPD fragment patterns. The polymorphic levels of RAPD bands by some primers (OPA-2,4,3,9,10,20) were very high in all 29 strains, with 3,030 fragments ranging between 200 and 2,000 bp. Intraspecific genetic dissimilarity of the 29 strains was calculated to range from 3.3% to 45% by Nei-Li's method using these 3,030 RAPD bands. The genetic variation among Korean strains was relatively high, with dissimilarities ranging between 17% and 38.6%. In the Neighbor-Joining analysis using the genetic dissimilarities based on RAPD, all 29 strains were classified into 5 clusters. Strains in each cluster showed specific characteristics according to their origin and strains. These results suggested that OPA and OPB primers could be used for developing molecular genetic markers and screening of unidentified (F. velutipes) strains.

• Journal of Life Science
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• v.20 no.12
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• pp.1743-1749
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• 2010

A research group demonstrated that the 37 kDA protein of Edwardsiella tarda, a causing causative agent of edwardsiellosis in fish, exhibited high antigenicity in Japanese flounder. The research group also showed that the N-terminus amino acid sequences of the 37 kDa protein were mapped to the N-terminus of GAPDH (glyceraldehyde-3-phosphate dehydrogenase). Using degenerated primer sets based on the known N-terminus sequence, the corresponding E. tarda DNA was amplified and cloned. The nucleotide sequences of the cloned gene revealed high homology with a bacterial gene for GAPDH, as we was expected. The amino acid sequence of E. tarda GAPDH (etGAPDH) revealed a <70% similarity with GAPDH proteins in other Enterobacteriaceae. With the application of artificial protein overexpression system in Escherichia coli, the recombinant etGAPDH (rGAPDH) was produced and purified. In this study, Using the purified rGAPDH, the etGAPDH specific polyclonal antibody has been was generated using the purified rGAPDHin this study. The immunoblotting analyses demonstrated that the location of the GAPDH protein is located with the association of is associated with the envelops of E. tarda. The rGAPDH was administrated into Japanese flounder via IP route for evaluation of the protective ability. Although the specific antibody titer against etGAPDH was increased about 3-fold after 4 weeks post-vaccination, the survival rates of vaccinated Japanese flounder and the control group with wild type E. tarda was were 12.5% and 0%, respectively. Our results indicated that rGAPDH is immunoreactive antigen but that it will not generate protective immunity in Japanese flounder.