• Korean Journal of Microbiology
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• v.32 no.3
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• pp.198-201
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• 1994

The nucleotide sequence of the cohesive end site (cos) of Lactobacillus casei phage J1 genome was determined. Comparison between the nucleotide sequences of the circular cos and the left end of the linear J1 DNA showed that the nicking sites of the terminase were as follows: 5'- GGTCGGCC$\downarrow$ -3' 3'- $\uparrow$CCAGCCGG -5' The cohesive single-stranded ends of J1 were found to be 3'-protruding and composed of 8 nucleotides. The mol% G + C of the cohesive ends was 87.5. The cos site shows dyad symmetry from -33 to + 25 bp if the 5' terminal nucleotide of the left end of the linear J1 DNA is numbered +1. No homology was found among the cos sites of phages reported so far.

• The Plant Pathology Journal
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• v.18 no.4
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• pp.187-191
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• 2002

This paper describes the cloning and sequence analysis of the 5'-terminal region and full-length cDNA production of genomic RNA of Lily symptomless virus (LSV), a Species Of the genus Carlavirus. A sing1e DNA band about 600 bp harboring the 5'-end of genomic RNA of the virus was successfully amplified by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE), and was cloned for nucleotide sequence determination. Sequence analysis of selected RACE cDNA clones revealed that the LSV 5'non-translated region consists of 67 nucleotides long of AT rich stretch followed GC rich from the 5'-end. To produce full-length cDNA products for the viral genomic RNA, a set of LSV-specific primers could be designed based on the obtained sequence in this study and the known sequences of 3'-terminal region for the virus. Full-length cDNA copies of LSV, an 8.4 kb long, were directly amplified by the long-template RT-PCR technique from the purified viral genomic RNA samples. This full-length cDNA copies were analyzed by restriction mapping. The molecules produced in this study can be useful for the production of in vitro infectious cDNA clone, as well as, for the completion of genomic RNA sequence and genome structure for the virus.

• Korean Journal of Food Science and Technology
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• v.24 no.1
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• pp.59-64
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• 1992

Salted and then washed flounder muscles with salting levels of 10%, 15% and 20% were mixed with boiled foxtail millet and spices(radish, garlic, ginger and red pepper) and fermented at $15^{\circ}C$ for 7 days. The changes of taste constituents of fermented flounder Sikhae, such as sugars, free amino acids and 5'-nucleotides, were investigated. The content of fructose decreased significantly during Sikhae fermentation, but the content of mannitol that was not detected from raw material was estimated to be $6.26%{\sim}8.97%$ in Sikhae. The content of total free amino nitrogen in the 15% salted Sikhae was 290.6 mg% and the highest value with 53.4% of its extract nitrogen. It is believed that leucine, alanine, arginine, glutamine, isoleucine, valine, glutamic acid and lysine may play an important role as the taste constituents in Sikhae. The detected 5'-nucleotides were CMP, UMP, CTP, AMP, ADP and ATP and among them the nucleotide showing the hightest level irrespective of treatment was UMP estimated to be $761.0\;{\mu}g{\sim}849.0\;{\mu}g/g$. ATP and ADP were significantly decreased in Sikhae, but CMP and CTP were significantly increased in the 15% salted Sikhae compared with those of raw material.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.2
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• pp.175-179
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• 1999

Processing conditions for dried condiments with oyster, pen shell and cockle shell were investigated. The enzymatic hydrolysis for 3 hours was more profitable than hydrothermal extraction to develop flavoring matters from oyster, pen shell and cockle shell. As a result of omission tests, nucleotides were predominated in the taste compounds of shellfish hydrolysates rather than free amino acids, and the contribution of nucleotides and free amino acids to the taste of shellfish hydrolysates was remarkable. The major flavoring components of shellfish hydrolysates were free amino acids and oligopeptides below 500 dalton. When shellfish hydrolysates were separated with membrane (molecular weight cutoff 500 dalton) for recovering flayer, recovering yields of amino type nitrogen were $92.1\~92.8\%$. Moisture contents of dried shellfish condiments prepared with pretense hydrolyzed oyster, pen shell and cockle shell were $3.5\%,\;3.8\%$ and $3.7\%$, respectively. Contents of total nitrogen were $69.4\%,\;78.8\%$ and $74.2\%$, and those of amino nitrogen were $45.5\%,\;48.9\%$ and $45.4\%$, respectively. Drying yield, solubility and absorption rates at Aw 0.88 were $11.7\%,\;78.4\%$ and $6.8\%$ in oyster, $8.2\%,\;73.6\%$ and $6.1\%$ in pen shell, $9.8\%,\;76.9\%$ and $6.6\%$ in cockle shell, respectively.

• Journal of Applied Biological Chemistry
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• v.44 no.4
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• pp.163-166
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• 2001

Piezoelectric (PZ) crystal biosensor system was used to detect the DNA of food pathogenic Listeria monocytogenes. L. monocytogenes-specific DNA was multiplied via the polymerase chain reaction using LM1 oligonucleotide (5'-TTACGAATTAAAAAGGAGCG-3') and LM2 oligonucleotide (5'-TTAAATCAGCAGGGGTCTTT-3') as primers. DNA fragment of 161 bp, which was specific only for L. monocytogenes, was observed. To obtain a large amount of single-stranded DNA containing an SH group used for coupling to the gold electrode chemisorptively, LM1 oligonucleotide containing a mercaptohexyl group was utilized as a single strand PCR primer. The PCR product was immobilized onto the gold electrode of PZ crystal, and hybridization was monitored in quartz crystal microbalance (QCM) system by injecting the antisense single-stranded DNA of 161 nucleotides obtained via the single strand PCR using the unmodified LM2 primer. Approximately 70 Hz of frequency drop was observed in the QCM system in the case of two consecutive injections of $5{\mu}g$ of the antisense single-stranded DNA.

• Journal of Veterinary Science
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• v.22 no.6
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• pp.66.1-66.9
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• 2021

Background: Maedi/Visna virus (MVV) is a contagious viral pathogen that causes considerable economic losses to the sheep industry worldwide. Objectives: In China, MVV has been detected in several regions, but its molecular characteristics and genetic variations were not thoroughly investigated. Methods: Therefore, in this study, we conducted next-generation sequencing on an MVV strain obtained from northwest China to reveal its genetic evolution via phylogenetic analysis. Results: A MVV strain obtained from Inner Mongolia (NM) of China was identified. Sequence analysis indicated that its whole-genome length is 9193 bp. Homology comparison of nucleotides between the NM strain and reference strains showed that the sequence homology of gag and env were 77.1%-86.8% and 67.7%-75.5%, respectively. Phylogenetic analysis revealed that the NM strain was closely related to the reference strains isolated from America, which belong to the A2 type. Notably, there were 5 amino acid insertions in variable region 4 and a highly variable motif at the C-terminal of the surface glycoprotein (SU5). Conclusions: The present study is the first to show the whole-genome sequence of an MVV obtained from China. The detailed analyses provide essential information for understanding the genetic characteristics of MVV, and the results enrich the MVV library.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.173-178
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• 1999

The extracellular cytosine deaminase (EC 3.5.4.1) from Chromobacterium violaceum YK 391 was purified 264.7-fold with an overall yield of 14.3%. The enzyme was for the first time homogeneous by the criteria of polyacrylamide gel electrophoresis performed in the absence and in the presence of sodium dodecyl sulfate. The molecular weight of the purified enzyme was estimated to be about 156 kDa. The enzyme consisted of two identical subunits of approximate molecular weight 78 kDa. The isoelectric point of the enzyme was pH 5.55. The enzyme had a pH optimum of 7.5 and a temperature optimum of around 40 to $45^{\circ}C$. Besides cytosine, the enzyme deaminated 5-fluorocytosine, cytidine, 5-methylcytosine, and 6-azacytosine, but not 5-azacytosine. The extracellular cytosine deaminase is believed to be unique because it was active not only on cytosine but also on cytidine. The apparent $K_m$ values for cytosine, 5-fluorocytosine, cytidine, and 5-methylcytosine were determined to be 1.55 mM, 5.52 mM, 10.4 mM, and 67.2 mM, respectively. The enzyme activity was strongly inhibited by heavy metal ions such as $Fe^{2+},Pb^{2+},Cd^{2+},Zn^{2+}, Hg^{2+}, and Cu^{2+}$ at 1 mM, and completely by $\alpha,\alpha$'-dipyridyl, and $\rho$-chloromercuribenzoate at 1 mM, and weakly inhibited by 1mM ο-phenanthroline. The enzyme activity was not affected by various nucleosides and nucleotides.

• Parasites, Hosts and Diseases
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• v.40 no.3
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• pp.131-138
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• 2002

The cyclophilins (Cyps) are family members of proteins that exhibit peptidylprolyl cis-trans isomerase (PPIase, EC 5.2.1.8) activity and bind the immunosuppressive agent cyclosprin A (CsA) in varying degrees. During the process of random sequencing of a cDNA library made from Giardia intestinalis WB strain, the cyclophilin gene (gicypl) was isolated. An open reading frame of gicyp1 gene was 576 nucleotides, which corresponded to a translation product of 176 amino acids (Gicypl). The identity with other Cyps was about 58-71%. The 13 residues that constituted the CsA binding site of human cyclophilin were also detected in the amino acid sequence of Gicypl, including tryptophan residue essential for the drug binding. The single copy of the gicypl gene was detected in the G. intestinalis chromosome by southern hybridization analysis. Recombinant Gicyp 1 protein clearly accelerated the rate of cis ${\rightarrow}$ trans isomerization of the peptide substrate and the catalysis was completely inhibited by the addition of $0.5{\;}{\mu}M$ CsA.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.5
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• pp.512-518
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• 2002

In order to effectively utilize of granular ark and ark shell, lipid and fatty acid compositions, free amino acids, nucleotides and their related compounds and minerals in the muscle and viscera of raw and cooked specimens were analyzed. The major constituents of non-polar lipids in the granular ark and ark shell were triglycerides, which showed higher content in viscera than the muscle. The polar lipids in the granular ark and ark shell were mainly consisted of phosphatidylcholine and phosphatidylethanolamine. The major fatty acids of total lipid were 16:0, 20:5n-3, 18:1n-9, 16:1n-7, 18:0 and 22:6n-3 both the granular ark and ark shell. The major nucleotides and the related compounds were adenosine monophosphate and adenosine diphosphate and they had higher content in the muscle than in viscera both samples, free amino acids such as taurine, glycine, alanine, glutamic acid, phenyl alanine and aspartic acid were abundant both the granular ark and ark shell. In the raw muscle of granular ark, glycine, alanine and $\alpha$-amino-iso-butyric acid were high level, but glutamic acid, aspartic acid and phenyl alanine were low level compared with those of cooking muscle. In the raw muscle of ark shell, taurine and $\alpha$-amino-iso-butyric acid were high content, but the glutamic acid and aspartic acid were low level compared with those of cooking muscle. Minerals in the granular ark and ark shell were chiefly consisted of potassium, sodium, magnesium, iron and calcium.

• Journal of Nutrition and Health
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• v.11 no.3
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• pp.13-20
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• 1978

Changes of free amino acids, nucleotides and their related compounds as taste compounds during sun drying of ascidian Cynthia roretzi, were analyzed by amino acid autoanalyzer and high speed liquid chromatography. In fresh ascidian, the results showed that 5'-UMP $(12.1\;{\mu}mole/g)$ was dominant and the content of cytosine, 2', 3'-CMP, 2', 3'-GMP, hypoxanhtine, 5'-AMP,5'-IMP were 5.8, 3.4, 3.1, 2,3, 1.7 and $1.3\;{\mu}mole/g$ ondry base respectively. 5'-IMP, 2', 3'-CMP and 2', 3'-GMP tended to degrade slowly and 5'-AMP, cytosine and 5'-UMP were decreased rapidly while hypoxanthine were increased remarkably during the sun drying. In dried ascidian, the content of hypoxanthine was the highest, 7.2 mole/g on dry base, whereas that of 5'-AMP $(0.5\;{\mu}mole/g)$) and 5'-IMP $(0.9\;{\mu}mole/g)$ were lower. Glutamic acid, alanine and serine were dominant amino acid in the fresh extracts, having 22.4% (611.3mg%, on dry qase), 19.8% (540.5mg%) and 14.8% (402.8mg%) of the total amino acid content respectively. The content of tyrosine, histidine, lysine, methionine, isoleucine and valine were low, and proline, phenylalanine were detected in trace amount. The free amino acid were not changed in composition but the increase of total free amino acid was approximately 116.8mg% during sun drying. In sun dried ascidian, glutamic acid (691.0mg, on dry base), alanine (641.3mg%), serine (469.5mg%), threonine (234.8mg%) and glycine (206.3mg%) were dominant amino acid. It is believed that glutamic acid, serine, alanine, threonine, glycine and hypoxanthine play an important role as taste compounds in sun dried ascidian.