• 한국식품과학회지
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• 제32권6호
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• pp.1331-1335
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• 2000

김치 젖산균인 WL6는 API kit 또는 Biolog system방법에 의하여 동정한 결과, Leuconostoc mesenteroides ssp. cremoris, Leu. mesenteroides ssp. dextranicum 또는 Lactobacillus bifermentans로 나타나 동정되지 않았다. 그러나, pepN gene과 16S rRNA gene으로부터 2개의 specific-sequence primer set을 제조하여 PCR 방법으로 증폭한 후에 표준균주들과 비교한 결과, WL6는 Lactobacillus bifermentans로 추정되었다.

• The Plant Pathology Journal
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• 제22권2호
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• pp.155-160
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• 2006

A strain of Pepper mottle virus (PepMov) was isolated from chili pepper plants in Korea. In host range study, this virus, designated PepMoV-SNU1, shared most characteristics with PepMoV isolates reported previously. Thermal inactivation point ($45^{\circ}C\;to\;75^{\circ}C$) and dilution end point ($10^{-1}\;to\;10^{-4}$) of PepMoV-SNU1 showed differences depending on the propagation hosts. Cylindrical and pinwheel-shaped inclusions were always observed in pepper leaf tissues infected with the virus alone. Unexpectedly, a special structure of pinwheel shaped inclusion surrounded with unknown small spots was also observed in the leaf section when co-infected with a strain of pepper mild mottle virus. The partial sequence of coat protein gene and 3' untranslated region of PepMoV-SNU1 showed 98% identity with those of other PepMoV isolates. A primer pair derived from 3' end of the coat protein gene and poly A tail regions were designed. Optimal detection condition of PepMoV-SNU1 by RT-PCR was tested to determine appropriate annealing temperature and additional volumes of oligo-dT (18-mer), dNTP, and Taq polymerase. Under the optimized condition, an expected 500 Up PCR-product was detected in pepper leaves infected with PepMoV-SNU1 but not in healthy plants.

• Clinical and Experimental Reproductive Medicine
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• 제24권1호
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• pp.51-56
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• 1997

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'dysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.109-114
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• 1996

General PCR technique alone has a limitation for preimplantation genetic diagnosis(PGD) using single blastomere. Recntly developed primer extension preamplification(PEP) technology amplifies the whole genome and thus, simultaneous multiple locus analysis became possible. In this study, we report the efficacy of PEP-PCR for PGD in three muscular dystrophy carriers undergoing IVF-ET. A total of 37 blastomeres were obtained from 40 embryos at six to eight cell stage in three IVF cycles in two DMD and one BMD carriers. Whole genome from single blastomeres were amplified using I5-base oligonucleotide random primers. PCR amplified products of exon 45 in the dystrophin gene and alphoid X/Y loci for gender determination were analysed by 2% metaphor gel electrophoresis. A total of 37 PEP-PCR replicates from 37 single blastomeres from 40 embryos and 37 blanks were performed. We obtained the reliable results for exon 45 and alphoid X/Y. Transfer of female embryos and unaffected male embryo was attempted in three couples. Unfortunately, pregnancy was not achieved in these cases. PEP-PCR is a reliable and efficient PGD method in multiple locus analysis using single blastomere.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1629-1633
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• 2015

Peptide assimilation in Helicobacter pylori necessitates a coordinated working of the peptide transport systems (PepTs) and aminopeptidase (PepA). We found that H. pylori hydrolyzes two detector peptides, _L-phenylalanyl- _L-3-thiaphenylalanine (PSP) and _L-phenylalanyl- _L-2-sulfanilylglycine (PSG), primarily before intake and excludes their antibacterial effects, whereas Escherichia coli readily transports them with resultant growth inhibition. PSP assimilation by H. pylori was inhibited by aminopeptidase inhibitor bestatin, but not by dialanine or cyanide-m-chlorophenylhydrazone, contrary to that of E. coli. RT- and qRT-PCR analyses showed that H. pylori may express first the PepTs (e.g., DppA and DppB) and then PepA. In addition, western blot analysis of PepA suggested that the bacterium secretes PepA in response to specific inducers.

• 식물병연구
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• 제23권2호
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• pp.193-201
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• 2017

국내의 토마토에서 발생하는 주요 바이러스는 Tomato chlorosis virus (ToCV), Tomato spotted wilt virus (TSWV), Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Tomato mosaic virus (ToMV)이다. 이들 바이러스를 진단하기 위해 프라이머 세트와 반응액을 포함하는 역전사중합반응(RT-PCR)법의 조건을 조사하였다. 공시한 바이러스에 특이적인 염기서열로부터 모두 46개 프라이머 세트를 설계하고, 이를 이용해 주형을 넣지 않은 RT-PCR에서 비특이 반응을 조사하였다. 이들 가운데 16개 조합을 건전한 토마토 RNA에 적용한 결과 프라이머 세트와 RT-PCR 반응액 간의 친화성이 비특이 반응 감소에 영향을 주었다. cDNA 합성과 관련된 인자와 RT-PCR 반응액 사이의 조합을 근거로 ToCV 진단을 위한 두 종류의 반응액을 선발하였다. ToCV 진단 시 수립된 조건을 나머지 바이러스 진단에 적용했을 때, 특이성이 높은 프라이머 세트 C029 (ToCV), C072 (TSWV), C070 (CMV), C048 (PepMoV), C065 (ToMV)를 선발할 수 있었다. 이들 프라이머 세트는 공시한 바이러스를 특이적으로 진단하는 데 유용할 것으로 판단된다.

• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.147.2-148
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• 2003

Complete nucleotide sequence of genome RNA of a Korean isolate of Pepper mottle virus (PepMoV-Vb) from field-collected diseased paprika (Capsicum annuum var grossum) was determined in this study. Symptoms of isolates of PepMoV were divided largely into two groups, vein banding (Vb) and vein clearing (Vc) patterns. PepMoV-Vb RNA consists of 9,640 nucleotides excluding the poly(A) tail. A single open reading frame was identified beginning at nucleotide position 169 encoding a polyprotein of 3024 amino acids which is typical of the Potyvirus genus. The complete nucleotide sequence and coding regions of PepMoV-Vb were compared to that of 11 potyviruses within the genus Potyvirus. The overall nucleotide sequence identity was 94.7 and 94.1％ identical to PepMoV-C and PepMoV-FL, respectively. Full-length cDNAs of PepMoV-Vbl were synthesized from purified viral RNAs by RT-PCR and their genome structure was analysed by RFLP analysis. This is the first report on complete nucleotide sequence of PepMoV isolated from paprika in Korea.

• 식물병연구
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• 제12권2호
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• pp.91-98
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• 2006

Acinetobacter sp. KTB3의 배양여과액으로 제조한 분말 제제 KNF2022의 바이러스 감염억제 효과를 PMMoV 와 N. glutinosa 또는 N. tabacum cv. Xanthi nc를 이용하여 검정한 결과, 증류수 1/100 배양액의 실질 농도 10,000 ppm) 희석농도에서 $94.3{\sim}95.6%$의 높은 감염억제 효과를 나타냈다. 이 1/100 희석농도를 이용하여 PepMoV-C amaranticolor에서 검정한 억제효과는 97.1%, CMV-C amaranticolor 에서는 92.5%의 감염억제를 나타냈다. KNF2022 희석액을 N. glutinosa의 반엽에 도말하고 일정 시간 경과 후 PMMoV를 접종하여 조사한 억제효과의 지속성은 처리 2 일 후까지 약 80% 선에서 억제효과를 나타냈으나, 처리 4 일 후부터는 50% 수준으로 저하되었다. 고추에 KNF2022를 처리한 후 3 종의 바이러스를 접종하고 발현되는 병징을 조사한 결과, 각 바이러스를 단독으로 접종하였을 경우는 바이러스의 종류에 관계없이 접종 후 10 일까지 병징발현이 지연되는 효과를 보였다. 특히 PepMoV를 접종한 경우는 접종 후 30 일까지 병징발현이 억제되어 제제의 처리효과가 뚜렷하게 나타났다. 한편 이들 3 종 바이러스의 복합감염에 대한 KNF2022의 효과는 모든 바이러스 조합에서 단독감염 보다 강하게 발현되어 병징억제의 효과는 뚜렷하게 나타나지 않았다. 제제를 처리한 고추에서 증식된 바이러스의 농도를 PCR 및 ELISA로 검출한 결과, PepMoV를 단독으로 접종한 경우, 접종 후 30 일까지 cDNA가 검출되지 않았다. RT-PCR 검정에서 억제효과가 인정된 PepMoV와 그 복합감염 의 조합에 대해서 ELISA 검정을 실시한 결과, PCR 검정에서 얻어진 결과와 유사한 패턴을 보여 PepMoV에 대한 감염억제효과가 뚜렷하게 인정되었다. KNF2022를 처리하여 전자현미경으로 관찰한 PMMoV와 PepMoV는 처리 5 분 후에 각각 200-250 nm 및 400-600 nm의 길이로 절단된 입자가 많이 관찰되었고, 처리 30 분 후에는 대부분 100-150nm와 300-500 nm의 절편입자로 관찰되었다. 이와 같이 절단된 입자가 증가할수록 N. tabacum cv. Xanthi nc 와 C. amaranticolor에서 나타난 병반수는 급격하게 감소되었다. 한편 KNF2022를 처리한 후 전자현미경을 관찰한 CMV의 바이러스 입자는 처리시간에 비례하여 파괴된 입자의 수가 증가하였으며, 파괴된 바이러스입자가 증가할수록 C. amaranticolor에서의 병반수도 감소되었다.

• Journal of Microbiology and Biotechnology
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• 제20권12호
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• pp.1769-1771
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• 2010

The envelope glycoprotein E2 of hepatitis C virus (HCV) binds to various cell surface receptors for viral infection. We performed biopanning against this protein and selected peptides from phage display peptide libraries. Two short peptides, pep7-1 and pep12-1, were selected and their ability to inhibit the infection process was investigated. When pep7-1 was present, the infectivity of HCV particles in cell culture was notably decreased. This decrease was demonstrated by Western blot analysis, immunofluorescence assay, and reverse transcription PCR assay. However, pep12-1 showed little inhibitory effect on HCV infection.

• Journal of Genetic Medicine
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• 제2권1호
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• pp.35-39
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• 1998

Duchenne/Becker muscular dystrophy are the major neuromuscular disorders with X-linked recessive inheritance. Preimplantation diagnosis of sex determination has been generally used to avoid male pregnancies with these diseases. However, in order to determine if the embryo is normal, carrier or affected regardless of the sex, there is a need for a combined analysis of specific exon on dystrophin gene as well as sex determination of embryo using the same biopsied blastomere. If the exon deletion is not determinable, further diagnosis of carrier or patient can be performed by haplotype analysis. In this study, we applied the primer extension preamplification (PEP) method, which amplifies the whole genome, in 40 cases of single amniocyte and 40 cases of chorionic villus cell. We analysed haplotypes using two (CA)n dinucleotide polymorphic markers located at the end of 5' and 3' region of the dystrophin gene. Exon 46 of dystrophin gene and DYZ3 on chromosome Y were chosen as a target sequence for coamplification PCR. Upon optimizing the conditions, the amplification rates were 91.25% (73/80) for haplotypes (92.5% in amniocyte, 90% in chorionic villus cell) and 88.75% (71/80) for coamplification (85% in amniocyte, 92.5% in chorionic villus cell). The result of the study indicates that haplotypes analysis and coamplification of dystrophin and Y-specific gene using PEP can be applied to prenatal and preimplantation diagnosis in Duchenne/Becker muscular dystrophy making it possible to determine if the fetus is a carrier or an affected one.