• Microbiology and Biotechnology Letters
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• v.49 no.3
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• pp.316-319
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• 2021

Restriction endonucleases play an important role in molecular cloning, clinical diagnosis, and pharmacological drug studies. In this study, DNA-templated copper nanoclusters (DNA-CuNCs) were used to detect AluI endonuclease activity due to their high fluorescence emission and rapid synthesis of DNA-CuNCs under ambient conditions. Results showed that AluI activity was detected in a highly sensitive manner at low concentrations of AluI endonuclease by the fluorescence intensity of DNA-CuNCs. Additionally, its inhibition was monitored in the presence of daidzein under optimal conditions.

• Korean Journal of Biological Psychiatry
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• v.23 no.4
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• pp.140-147
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• 2016

Objectives To determine the relationship between the Alu insertion/deletion (I/D) polymorphism in the tissue-type plasminogen activator (tPA) gene and the clinical outcome of mirtazapine treatment in Korean major depressive disorder (MDD) patients. Methods We enrolled 422 patients in this study. Symptoms were evaluated using the 21-item Hamilton Depression Rating (HAMD-21) Scale. After 1, 2, 4, and 8 weeks of mirtazapine treatment, the association between the Alu I/D polymorphism in the tPA gene and remission/response outcomes were evaluated. Results The proportion of I/I homozygotes in responders was higher than that in non-responders, whereas the proportion of D/D homozygotes in responders was lower than that in non-responders at 8 weeks of treatment (p = 0.032, OR = 1.57). The percentage decline of HAMD-21 scores in I allele carriers was larger than that of D/D homozygotes at 2 and 8 weeks of treatment (p = 0.035 and 0.007, respectively). I allele carriers were associated with remission at 8 weeks of treatment (p = 0.047, OR = 2.2). Conclusions These results show that treatment response and remission to mirtazapine were associated with the Alu I/D polymorphism of the tPA gene. This suggests the Alu I/D polymorphism may be a potential genetic marker for the prediction of therapeutic response to mirtazapine treatment in patients with MDD.

• Microbiology and Biotechnology Letters
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• v.38 no.4
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• pp.455-460
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• 2010

A polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis was applied to detect and identify ten Weissella spp. frequently found in kimchi. The previously reported genus-specific primers designed from 16S rDNA sequences of Weissella spp. were adopted but PCR was performed at the increased annealing temperature by $4^{\circ}C$. The sizes of amplified PCR products and restricted fragments produced by AluI, MseI, and BceAI endonucleases were well correspond with the expected sizes. W. kandleri, W. koreensis, W. confusa, W. minor, W. viridescens, W. cibaria, and W. soli were distinguished by AluI and MseI and W. hellenica and W. paramesenteroides were identified by BceAI. W. thailandensis was distinguished when restriction pattern of other species was compared but identified by the single use of MspI.

• Proceedings of the Korean Institute of Communication Sciences Conference
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• 2006.07a
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• pp.423-423
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• 2006

• Journal of Life Science
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• v.8 no.2
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• pp.126-130
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• 1998

A pair of designed primers (sequences from Gene Bank) amplified 16S fRNA gene of V. vulnificus within polymerase chain reaction (PCR) machine. This PCR product is about 1.3kb DNA fragment. Six enzymes (BamH I, Alu I, Sau3A I, Hind III, Sal I, Sma I) were used for restriction pattern analysis of amplified 16S rRNA gene of V. vulnificus ATCC 27562. Digested fragments are resolved by 3% agarose gel. BamH I did not show digested fragment so, there was no cutting site of BamH I in PCR product. Alu I produced three small fragments from 400 bp to 200 bp. Sau3A I produced three fragments larger than Alu I from 70 bp and 500 bp. One of fragments of Sal I was same with 500 bp of Hind III fragment and the other was 750 bp. Sma I showed two fragments of 800 bp and 470 bp. The profile of digested fragments of 16S rRNA of V.vulnificus ATCC 27562 will may be able to use standard profile for identification of V. vulnificus.

• Journal of Embryo Transfer
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• v.28 no.4
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• pp.355-360
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• 2013

Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.

• Ocean and Polar Research
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• v.26 no.4
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• pp.551-557
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• 2004

Polymorphism of the lipoprotein lipase (LPL) gene which plays an important role in regulation of lipid deposition was analysed in two red seabream (pagrus major) populations (KF4, cultured KORDI line, n=100 : JPN, imported from Japan, n=100). We amplified a DNA fragment (1,091 bp) including the exon 2 region of the LPL gene, and conducted PCR-RFLP analysis using MspI and AluI. The PCR products were also sequenced. Two alleles (A and B) were found in MspI digestion and Sve alleles (A, B, C, D and E) in AluI digestion. The sequenced data revealed four nucleotide substitutions including one transversion at the MspI recognition site (nt 2,235, $C{\rightarrow}10$) and three transitions at the AluI recognition sites (nt 1,721, $A{\rightarrow}G;$ nt 2,319, $C{\rightarrow}T;$ nt 2,319, $T{\rightarrow}C$). Among them, substitutions at the nt 2,235 and 2,319 sites which are located in the exon 2 were proved to be silent point mutations. MspI polymorphism resulted in 3 genotypes, and the allele frequency was significantly different between the two fish populations, KF4 and JPN. In the case of AluI polymorphism, the 5 alleles (A, B, C, D, E) comprised 12 genotypes of the 5 alleles. KF4 population, alleles D and I were specific to the LPL gene Polymorphisms would be useful DNA markers for red seabream population.

• Proceedings of the Korean Aquaculture Society Conference
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• 2003.10a
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• pp.33-33
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• 2003

Lipoprotein lipase (LPL)은 지방축적과 대사에 있어 중요한 효소로서 지방조직과 근육 내로의 지방산 유입을 조절하며, 여러 조직에서 합성되고, 조직 특이적인 방법으로 동물의 생리상태, 영양상태 및 발달단계에 따라 유전자 발현이 조절되는 것으로 알려져 있다. 본 연구는 어류의 체지방 축적과 대사과정을 이해하고 성장 및 경제형질 관련 DNA marker를 확보하기 위한 1차적인 연구로서, 한국산 선발계통 및 일본산 양식계통과 이들 두 계통간 잡종 참돔 집단을 이용하여 LPL 유전자 내의 다형성을 탐색하고 분석하였다. PCR-RFLP 분석을 실시하여 참돔 LPL 유전자 exon 2번을 포함하는 영역에서 3개의 (Msp I, Alu I 및 Hsp92II) 다형성을 확인하였고, 각 집단간 대립유전자의 빈도를 분석하였다. Exon 2번에서 관찰된 Msp I 다형성은 염기치환 (C$\longrightarrow$G)이 일어난 형태로서 아미노산 서열에는 변화가 없는 silent mutation 이었으며, 대립유전자의 빈도를 분석한 결과, 각 집단간 뚜렷한 차이는 없었다. Alu I 및 Hsp92II 다형성은 intron 영역에서 발견되었으며, Alu I 다형성으로 인한 4개의 대립유전자형 중 D 대립유전자는 한국산 선발 계통과 한국산 선발계통을 포함하는 교배집단에서만 검출되었다. 이 후의 연구에서는 참돔 LPL 유전자의 exon 영역에 존재하는 다형성을 탐색하고, 형질과의 연관성 및 지방축적 기능과의 관련성 등을 분석하고자 한다.

• Journal of Life Science
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• v.19 no.9
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• pp.1328-1332
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• 2009

Anisakidosis is caused by anisakid nematodes (family Anisakidae) larvae which can cause not only direct tissue damage but also a severe allergic response related to excretory-secretion products. Lots of different species of anisakid larvae, including Anisakis simplex, Contracaecum, Goezia, Pseudoterranova, and Hysterothylacium, cause the anisakidosis. But it is difficult to diagnosis the species of larvae since the morphologies of larval anisakid nematodes are almost indistinguishable. In order to diagnosis the differential infections of larval anisakid nematodes, polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) of 18S rDNA - was conducted. Three major species of anisakid larvae including A. simplex, C.ontracaecum spp, and Goezia spp. were collected from mackerel (Scomber japonicus), mullet (Mugil cephalus), founder (Paralichthys olivaceus), eel (Astroconger myriaster) and red sea bream (Pagrus major). PCR amplified 18S rDNA from each species of anisakid larvae was digested with eight restriction enzymes including Taq I, Hinf I, Hha I, Alu I, Dde I, Hae III, Sau96 I, and Sau3A I. The original sizes of PCR amplified 18S rDNA were 2.0Kb in both anisakid larvaes and Goezia. Restrction enzymes including Hinf 1, Alu 1, Hha I, Dde 1 and Hae III cut differently and distinguished the A. simplex and Contracaecum type C'. However, Contracaecum type A showed two different restriction enzyme cutting patterns by Taq 1, Hinf I, Alu 1, and Dde 1. One of the patterns was the same as those of A. simplex, Contracaecum type C' and Goezia and the other was unique. These results suggest that PCR-RFLP pattern by Hinf 1, Alu 1, Hae I, Dde 1 and Hae III can be applied to differential diagnosis of human infection with A. simplex and Contracaecum type C'. Contracaecum type A needs further study of classification by morphological characteristics and genetic analysis.

• Interdisciplinary Bio Central
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• v.4 no.3
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• pp.8.1-8.7
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• 2012

The motor neuron degeneration 2 (mnd2) mice carry a point mutation of A to T nucleotide transversion at the serine 276 residue of high temperature requirement A2 (HtrA2), resulting in losses of an AluI restriction enzyme site (5'AGCT3') and the HtrA2 serine protease activity. Moreover, dysfunctions of HtrA2 are known to be intimately associated with the pathogenesis of neurodegenerative diseases, including Parkinson's disease. Thus, this mnd2 mouse is an invaluable model for understanding the physiological role of HtrA2 and its pathological role in neurodegenerative diseases. Nevertheless, many molecular and cellular biologists in this field have limited experience in working with mutant mouse models due to the necessity of acquired years of the special techniques and knowledges. Herein, using the mnd2 mouse model as an example, we describe easy-to-use standard protocols for web-based analyses of target genes, such as HtrA2, and a novel approach for simple and accurate PCR-AluI-RFLP-based genotype analysis of mnd2 mice. In addition, band resolution of AluI-RFLP fragments was improved in 12% polyacrylamide gel running in 1X Tris-Glycine SDS buffer. Our study indicates that this PCR-AluI-RFLP genotype analysis method can be easily applied by the molecular and cellular biologist to conduct biomedical science studies using the other mutant mouse models.