• Korean Journal of Microbiology
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• v.32 no.4
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• pp.258-263
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• 1994

Genomic Southern hybridization of Frankia EuIKl strain, a nitrogen fixing symbiont of Elaeagnus umbellate root nodules, with nifH,D of K. pneumoniae as a probe, showed that 3.2 Kb and 5.5 Kb of BamHI fragments and 15 Kb PstI fragment were strongly hybridized with the probe, indicating nifH,D are located on these fragments. Using the same probe, one clone(pEuNIF) was isolated from the genomic library constructed into pWE15 cosmid vector by colony hybridization. The 3.2 Kb and 5.5 Kb BamHI fragments of this clone were hybridized with the same probe and this result corresponds to the genomic Southern hybridization data. However, using nifH of Frankia FaCl strain as a probe, only the 3.2 Kb BamHI fragment showed hybridization signal. Amino acid sequence deduced from nucleotide sequence of 3' terminus of the 3.2 Kb and 5' terminus of the 5.5 Kb fragments showed that the former was highly homologous with that of ArI3 nifD from 182nd to 240th amino acids, while the latter was from 241st to 282nd amino acids. These results show that nifH and partial nifD sequences are located on the 3.2 Kb fragment and residual sequences of nifH on the 5.5 Kb fragment which is contiguous to the 3.2 Kb fragment.

• Journal of Nutrition and Health
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• v.30 no.3
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• pp.307-317
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• 1997

To determine the effects of endogenous and exogenous strogen on serum lipid levels, twenty nonsmoking healthy Korean women were participated in this experiment for 12 weeks. They were assigned to three groups : (1) eight women aged 22 to 30(yr) for the premenopausal(Pre) group, (2) eight, aged 49 to 60(yr) for the postmenoparusal(Pst) group, (3) four, aged 23 to 30(yr) for the oral contraceptive(OC) group which used triphasic OC formulation. Fasting blood samples representing every phase of the hormonal levels were obtained from the subjects of the Pre and the OC group. From the subjects of the Pst group, fasting blood samples were obtained once per three weeks for 12 weeks. All the serum data were adjusted for dietary effects, exercise, personality type and body mass index(BMI) by using analysis of covariation(ANCOVA). Serum lipid levels of the three groups were significantly different. While serum levels of triglycerides(TG)(p<0.0001), low density lipoprotein-chloesterol(LDL-C)/high density liporotein-cholesterol(HDL-C) ratio (LDC-C/HDL-C)(P<0.01) and total cholesterol (TC)/HDL-C ratio (TC/HDL-C)(P<0.001) were significatnly high in the Pst group, serum HDL-C(P0.001) level was significantly high in the Pre group. The OC group showed significantly low serum TC(P<0.0001) and LDL-C(P<0.0001) levels. There was no signidicant difference in the fluictuation of serum lipid levels during the menstrual cycle of the Pre group. However, in the OC group, serum TG level was significantly increased at phase 2(P<0.05) where exogenous estrogen administration was highest. Even though other serum lipid levels of the OC group were not significantly fluctuated according to the exogenous estrogen administration, there was a trend of increased levels of serum TC, LDL-C, LDL-C/HDL-C and TC/HDL-C and decreased level of HDL-C during the menstruation period. Also, serum TC level was high(P<0.005) and serum TG level was low (P<0.005) at the baseline of the OC group compared with the periods of OC administration. When screening and counseling the female population at risk for coronary heart disease(CHD), the result of this study suggest that in may be desirable to divide the population into several groups according to their personal physiological characteristics, such as age, OC administration, menstrual cycle and menopause, as well as general risk factors for CHD.

• Clinical and Experimental Pediatrics
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• v.45 no.12
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• pp.1585-1590
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• 2002

Rhizomelic chondrodysplasia punctata(RCDP) is a rare autosomal recessive disorder clinically characterized by symmetrical shortening of the proximal limbs, contractures of joints, a typical dysmorphic face, cataracts, and itchyosis. Patients with RCDP can be subdivided into three subgroups based on biochemical analysis and complementation studies. RCDP type I results from mutations in the PEX7 gene encoding the peroxisomal targeting signal type II(PST2) receptors and presents with both a defect in plasmalogen biosynthesis and phytanic acid oxidation. RCDP type II is deficient in the activity of dihydroxyacetonephosphate acyltransferase(DHAP-AT). RCDP type III is deficient in alkyl-dihydroxyacetonephosphate synthase(alkyl-DHAP). We report a case of RCDP type I which was confirmed with biochemical study, fibroblast culture, and gene study.

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.43-51
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• 1993

Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

• Korean Journal of Microbiology
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• v.34 no.3
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• pp.120-125
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• 1998

A plasmid pNPG contains a genomic DNA complementing npgA1 which is located on the left arm of linkage group I. It transformed Aspergillus nidulans at a high frequency. No abortive transformants were observed and the $Trp^+$ transformants were all $Npg^+$. The 10.4 kb Psti fragment of the genomic DNA was subcloned into pILJ16, which increased the transformation efficiency by more than 200-folds. The transformants were mitotically unstable and yielded $Arg^-$ conidia at the frequency of more than 80%. An additional gene cloned into the plasmid containing the fragment was always lost with $argB^+$ marker. These characteristics strongly indicate the possibility that the plasmids autonomously replicate. The full activity of enhanced transformation was retained on the 4.9 kb EcoRI-HaeIII fragment. The DNA segment was similar to AMA1 rather than ANS1 in function and designated AMA2.

• Microbiology and Biotechnology Letters
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• v.29 no.1
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• pp.18-24
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• 2001

Expression of the baculovirus major envelope glycoprotein gene(gp64) is regulated by transcription from botha early and late promoters. To develop a transient expression vector under the control of gp64 gene promoter, the gp64 gene of Bombyx mori nucleopolyhedrovirus-K1(BmNPV-K1) was characterized. The gp64 gene was local-ized at EcoR I-Pst I 7.38-kb fragment of the BmNPV-K1 genome. The EcorR 1-Pst I 7.38-kb fragment was cloned and the nucleotide sequence of 2,277 bases including the coding region of gp64 gene was determined. Based on these results, transient expression vector using gp64 gene promoter was constructed and named as pBm64. E.coli lacZ gene was introduced onto pBm64 as a reporter gene and expressed transiently in B. mori 5(Bm 5) cells. The expression vector transfected into the cells was maintained stably for 1 to 5 days. In order to confirm the expression of the reporter gene by gp64 promoter, recombinant virus was constructed. The recombinant virus has two independent transcription units in opposite orientations with two promoters; gp64 and polyhedrin gene promoters each initiating transcription of $\beta$-galactosidase and polyhedrin, respectively. Polyhedra formation and expression of $\beta$-galactosidase in Bm5 cells infected with the recombinant virus were observed with phase contrast microscope and in situ staining.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.9
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• pp.1226-1230
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• 2005

The present investigation was undertaken to study the genetic polymorphism of the DRB3 exon 2 in 75 crossbred cattle by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Five genotypes i.e. HaeIII-a, HaeIII-b, HaeIII-e, HaeIII-ab and HaeIII-ae were observed when the 284 bp PCR products were digested with HaeIII restriction enzyme. The corresponding frequencies of these patterns were 0.53, 0.04, 0.01, 0.38 and 0.04, respectively. Digestion with RsaI restriction enzyme resolved 24 different restriction patterns. The frequencies of these patterns ranged from 0.013 (RsaI-f, RsaI-k and RsaI-c/n) to 0.120 (RsaI-n). The results revealed that the crossbred cows belonged to the RsaI patterns namely b, k, l, a/l, d/s, l/n, l/o and m/n, whose corresponding frequencies were 0.027, 0.013, 0.040, 0.027, 0.040, 0.067, 0.027 and 0.067, respectively. Digestion of the 284 bp PCR product of DRB3.2 gene with PstI in the crossbred cattle did not reveal any restriction site. These results suggested the absence of the recognition site in some of the animals. These results also revealed that the crossbred cows studied were in homozygous as well as heterozygous condition. On the basis of the above results it can be concluded that the DRB3.2 gene was found to be highly polymorphic in the crossbred cattle population.

• Korean Journal Plant Pathology
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• v.11 no.2
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• pp.173-178
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• 1995

동물계에서 항바이러스와관련된 dsRNA 의존성 인산화 효소(PKR)의 유전자를 식물체에서 발현시킬 경우 PKR에 의한 단백질합성 및 식물바이러스의 증식조절 가능성에 대한 기초자료를 확보하기 위하여 사람에서 분리된 PKR cDNA를 Agrobacterium 방법에 의하여 연초식물체(Nicotiana tabacum cv. Xanthi-nc)로 형질전환시켰다. HindIII/PstI처리에 의해 얻어지는 약 1.8kb의 phPKR cDNA절편을 일련의 유전자 조작 방법을 통하여 식물발현벡터인 pBI121에 도입하여, p12168을 재조합하였다. 이를 A. tumefaciens LBA 4404에 형질전환시켜 연초식물체형질 전환에 이용하였다. 2mg/l BA와 0.5mg/l NAA가 포함되고 100$\mu\textrm{g}$/ml의 kanamycin이 첨가된 MS배지에서 shooting시킨 후 phytohormone이 첨가되지 않은 MS배지상에서 rooting을 시켜 형질전환 연초식물체를 얻었으며, 형질전환식물체는 정상식물체와 유사한 생육양상을 나타내었다. 형질전환식물체의 유전자도입은 hPKR cDNA의 전사부여는 RT-PCR 방법에 의하여 확인되었다.

• Animal Systematics, Evolution and Diversity
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• v.13 no.2
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• pp.73-81
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• 1997

한국산 조류 중 박새속(genus Parus)에 속하는 4종 Parus major wladiwostokensis (박새), P. ater amurensis(진박새), P. palustrius hellmayri(쇠박새) 및 P. varius varius(곤줄 박이)를 대상으로 이들의 계통적 유연관계를 구명하기 위하여 mtDNA분석을 실시하였다. 6 base를 인지하는 10개의 제한효소를 처리하여 얻어진 mtDNA의 크기는 16.6~17.0Kb였으며 Pst I과 Pvu II는 종간 차이가 뚜렷하였다. 각 종간의 절편양상을 비교하여 Parus속의 종 간 분화정도를 비교한 결과 P. m. wladiwostokensis와 P. a. amurensis의 유전적 근연관계 가 가장 가까웠고(p=0.073, F=0.294) P. a. amurensis와 P. v. varius는 비교적 현저한 유전 적 차이를 보였다.(p=0.119, F=0.143). Brown 등 (1979)의 공식을 이용하여 박새속 4종의 분 화시기를 추정한 결과 이들은 후기 선신세(Pliocene)와 초기 홍적세(Pleistocene) 사이에 분 화된 것으로 추정되었다.

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.167-172
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• 1988

A genetic map of the IncP-1 group plasmid pKU10 has been prepared through the construction of recombinant plasmids containing various fragments of pKU10. Phenotypic analysis of these derivatives has identified the location of genes encoding resistance to ampicillin, tetracyclin, and chloramphenicol. The region involved in conferring resistance to ampicillin was located around two PstI sites that are 1.0Kb apart. The tetracyclin resistance gene was mapped on the region of HindIII E fragment and a part of HindIII D fragment, and the determinant for chloramphenicol resistance gene was localized on HindIII D fragment.