• 농업과학연구
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• 제23권1호
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• pp.99-107
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• 1996

Eighteen nuclear probes were used to examine RFLP(restriction fragment length polymorphism) between six species of Artemisia spp. of Korea. Total DNA from six different species of Artemisia was separately cut with three restrict enzymes. The PstI enzyme was showed to reduce the variation of polymorphisms than the other two enzymes(EcoRl and BamHI). The genetic variation of polymorphism was similar between the Dhewegiki-ssug and Cham-ssug. RAPD analysis was applied to the same six species of Artemisia spp. in order to assess the degree of DNA polymorphism within the Artemisia genus. Six species of Artemisia were evaluated for variation using a set of 11 random 10-mer primers. Nine out of the eleven primers revealed scorable polymorphisms between six species of Artemisia spp. Genetic distances between each of the species were calculated and cluster analysis was used to generate a dendrogram showing phylogenetic relationships between them This result indicates that molecular markers will be more usable in intraspecific study of Artemisia spp. than isoenzyme markers.

• 농업과학연구
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• 제47권1호
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• pp.173-182
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• 2020

Human Astrovirus (HuAstV), known as a waterborne virus, is a group IV positive-sense single-stranded RNA that belongs to Astroviridae. The first outbreak of HuAstV was reported in England in 1975. HuAstV can exist not only among clinical patients but also in various water environments, such as water for agriculture and vegetables. For diagnosis of HuAstV from water samples, a polymerase chain reaction (PCR) system has been developed. However, the PCR-based diagnostic method has problems in field application, such as reaction time, sensitivity and specificity. For this reason, in this study we developed the loop-mediated isothermal amplification assay (LAMP) system, aimed specifically at HuAstV. Three prepared LAMP primer sets were tested by specificity, non-specificity and sensitivity; one LAMP primer set was selected with optimum reaction temperature. The developed LAMP primer set reaction conditions were confirmed at 62℃, and detection sensitivity was 1 fg/μL. In addition, restriction enzyme HaeIII (GG/CC) was introduced to confirm that the LAMP reaction was positive. As a result, selected LAMP primer set was 100 - 1000 times more specific, rapid, and sensitive than conventional-nested PCR methods. For verification of the developed LAMP assay, twenty samples of cDNA from groundwater samples were tested. We expect that the developed LAMP assay will be used to diagnose HuAstV from various samples.

• Journal of Plant Biology
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• 제35권2호
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• pp.165-171
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• 1992

rbcL 유전자 발현조절에 관한 연구의 일환으로 Cp DNA로부터 분리한 rbcL 유전자를 클로닝하였다. 옥수수의 엽록체로부터 DNA를 분리한 후 제한효소 BamHI으로 절단하여 rbcL 유전자가 포함된 BamHI 9 절편을 pUC19에 클로닝하여 재조합 플라스미드 pRLYS1을 만들었다. 쌀의 rbcL 유전자 일부를 probe로 사용하여 pRLYS1과 Southern hybridization한 결과와 제한효소 BamHI, HindIII, 그리고 PstI으로 절단된 pRLYS1 절편의 전기영동 결과로부터 재조합 플라스미드의 내부에 완전한 rbcL 유전자의 존재를 확인하였고 삽입방향을 결정하였다.

• The Plant Pathology Journal
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• 제19권3호
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• pp.159-161
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• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

• Parasites, Hosts and Diseases
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• 제48권4호
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• pp.297-301
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• 2010

Recently, emerging waterbome protozoa, such as microsporidia, Cyclospora, and Cryptosporidium, have become a challenge to human health worldwide. Rapid, simple, and economical detection methods for these major waterborne protozoa in environmental and clinical samples are necessary to control infection and improve public health. In the present study, we developed a multiplex PCR test that is able to detect all these 3 major waterborne protozoa at the same time. Detection limits of the multiplex PCR method ranged from $10^1$ to $10^2$ oocysts or spores. The primers for microsporidia or Cryptosporidium used in this study can detect both Enterocytozoon bieneusi and Encephalitozoon intestinalis, or both Cryptosporidium hominis and Cryptosporidium paNum, respectively. Restriction enzyme digestion of PCR products with BsaBI or BsiEI makes it possible to distinguish the 2 species of microsporidia or Cryptosporidium, respectively. This simple, rapid, and cost-effective multiplex PCR method will be useful for detecting outbreaks or sporadic cases of waterborne protozoa infections.

• Asian-Australasian Journal of Animal Sciences
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• 제14권12호
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• pp.1659-1664
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• 2001

Several studies have been done regarding carcass traits and growth in pigs. Recently, these have progressed to examine increases in economic traits, including meat quality and meat quantity, by using candidate genes. One of them is the pituitary-specific protein PIT-1, a member of the POU (Pit-Oct-Unc) family of transcription factors playing an important regulatory role in developmental processes. In addition, muscle development is known to be regulated in part by growth factors and cytokines locally produced. Therefore, studies were performed to analyze PIT-1 genotypes and serum cytokines (IGF-I, IGF-II, TGF-${\beta}1$, EGF, cortisol, DHEA-S, IL-2, and IL-6) in castrated male pigs for their possible involvement in the development of carcass traits. The genotypes of PIT-1 gene were analyzed by PCR-RFLP with MspI restriction enzyme. But, only CD and DD genotypes, not CC genotype, have been detected. Based on PIT-1 genotyping, a significant difference in EGF expression beween CD type (78.8 ng/ml) and DD type (46.0 ng/ml) was detected (p<0.05), whereas other cytokines did not show any statistical significance depending on PIT-1 genotypes. Collectively, these results suggest the possibility that EGF could affect the formation of carcass traits.

• Journal of mucopolysaccharidosis and rare diseases
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• 제2권1호
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• pp.5-7
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• 2016

Mucolipidoses II and III alpha/beta (ML II and ML III) are lysosomal disorders in which the essential mannose-6-phosphate recognition marker is not synthesized onto lysosomal hydrolases and other glycoproteins. The disorders are caused by mutations in GNPTAB, which encodes two of three subunits of the heterohexameric enzyme, N-acetylglucosamine-1-phosphotransferase ML II, recognizable at birth, often causes intrauterine growth impairment and sometimes the prenatal "Pacman" dysplasia. The main postnatal manifestations of ML II include gradual coarsening of neonatally evident craniofacial features, early cessation of statural growth and neuromotor development, dysostosis multiplex and major morbidity by hardening of soft connective tissue about the joints and in the cardiac valves. Fatal outcome occurs often before or in early childhood. ML III with clinical onset rarely detectable before three years of age, progresses slowly with gradual coarsening of the facial features, growth deficiency, dysostosis multiplex, restriction of movement in all joints before or from adolescence, painful gait impairment by prominent hip disease. Cognitive handicap remains minor or absent even in the adult, often wheelchair-bound patient with variable though significantly reduced life expectancy. As yet, there is no cure for individuals affected by these diseases. So, clinical manifestations and conservative treatment is important. This review aimed to highlight the extra-skeletal clinical problems in ML II and III.

• 대한의생명과학회지
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• 제8권4호
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• pp.275-282
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• 2002

This study was carried out to investigate the effect of fibrinogen genotype and other characteristics on the plasma fibrinogen levels. Many studies have conformed that high plasma fibrinogen levels are associated with the increased risk of ischaemic heart disease, stroke and arterial disease. And fibrinogen levels are related with age, obesity, cholesterol and alcohol consumption, genotypes. For this study the blood samples were collected from 93 healthy Koreans (66 males and 27 females). The blood samples were individually analyzed by smoking status, cholesterol levels, genotype, age, and gender. The plasma fibrinogen was assayed by clotting method (modified Clauss assay) and cholesterol was assayed by cholesterol oxidase method. Subjects were classified by current smokers, ex-smokers (<6 month), or nonsmokers. The $\beta$-fibrinogen genotype was detected by PCR of relevant region and digestion with HaeIII, with the H$_1$H$_1$ allele allowing cleavage by this restriction enzyme and H$_2$H$_2$ allele being refractory. In conclusion, the study shows that the factor of the increasement in the fibrinogen level was closely related with the cholesterol level, smoking status and genotype (H$_1$H$_2$); but there was no significant difference by gender, Especially, among the people over 50 years of age, fibrinogen level was higher with the increasement of cholesterol level (<200 mg/dl), current smoker, and genotype H$_1$H$_2$.

• Journal of Animal Science and Technology
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• 제55권6호
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• pp.551-557
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• 2013

This study was conducted to analyze the genotypes and genes of FABP3 (Fatty-acid Binding Protein 3) in pigs using MSPI restriction enzyme and electrophoresis. Analysis of data collected from a total of 210 crossbred pigs (LYD or YLD) in Chungcheongnam-do, Korea, revealed the following. The AA genotypes of FABP3 were detected in the 750 bp and 100 bp bands, while the Aa heterotype appeared in the 850, 750 and 100 bp bands and the aa recessive homotype was detected in a single band of 850 bp. The genotype frequency of AA, Aa and aa was 46.67%, 51.43% and 1.90%, respectively. The genetic equilibrium of this population showed a significant difference (p<0.001) based on a ${\chi}^2$-test. The carcass weight, backfat thickness, marbling score, pH, drip loss, cooking loss, and meat color based on the CIE $L^*$, and $b^*$ values according to genotypes of FABP3 did not differ significantly (p>0.05); however, the CIE $a^*$ values did (p<0.05).

• 대한바이러스학회지
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• 제29권2호
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• pp.99-105
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• 1999

BamHI restriction pattern and genomic library of Herpes simplex virus type 2 (HSV-2) strain G were constructed, and locations of the glycoproteins gB and gD, and tk genes on the fragments were detected by Southern blot analysis. HSV-2 genomic DNAs were cleaved into twenty-seven fragments by BamHI enzyme in the range of 0.72 to 15.08 (total 150.44 kb), which were cloned into the BamHI site of pBluescript SK(+) to construct genome library of the HSV-2. The library was named by the order of the fragment size from smallest one to largest one. HSV-2 glycoprotein gD gene was located in pHLA2-21 and pHLA2-22 recombinant plasmids, gB gene in pHLA2-24 plasmid, and tk gene in pHLA2-11 clone by Southern blot analysis.