• 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.

• 한국잠사곤충학회지
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• 제37권1호
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• pp.46-51
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• 1995

국내 분리주의 BmNPV를 이용하여 유용 단백질을 생산할 수 있는 새로운 전이벡터의 제작과 외내 유전자의 발현에 대한 연구결과는 다음과 같다. 1. PCR 기법에 의하여 다각체 단백질 유전자의 ＋1 - －194에 해당하는 promoter 부위를 증폭하고 클로닝 하였다. 2. 다각체 단백질 유전子의 5' 및 3'의 leader 부위를 promoter 부위와 함께 단계적으로 클로닝하여 전이벡터를 완성하였다. 3. 완성된 전이벡터는 pBmKSK1으로 명명하였으며, 염기서열 결정을 통하여 다각체 단백질 유전자의 ＋2 부터 ＋597까지 제거되고 외내 유전자의 클로닝 sites로 EcoRI, KpnI과 SacI을 가짐을 확인하였다. 4. 외래 유전자로서 E. coli의 $\beta$-galactosidase 유전자를 pBmKSKl에 클로닝하고 누에 세포주에 전이시킨 후, X-gal 염색 및 SDS-PAGE에 의하여 pB- mKSKl이 제 기능을 수행함을 확인하였다.

• 대한의생명과학회지
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• 제10권1호
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• pp.55-63
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• 2004

Bovine leukemia virus (BLV) is a causative agent for lymphoma disease in cattle including cows worldwide. BLV shares similar virion structure and characteristics with other retroviruses. The pol gene of the BLV genome produced reverse transcriptase (RT) and integrase (IN) for important roles for BLV genome integration into host cell chromosomes that is known to be coded in the 3' side of the BLV pol gene (one third portion). In this study, we have sequenced 978 bp in the 3' side of the BLV pol gene from BLV 10C3 in order to determine the BLV IN region of it. And we compared it to the nucleotide sequences of an Australian BLV isolate. As a result, nucleotide sequences of the IN region of the Korean-type BLV pol gene were mutated at a rate of 3.7%. We can confirm that the typical mutations are such as Arg (AGG) $\rightarrow$ Lys (AAG), Thr (ACG) $\rightarrow$ Met (ATG), Ile (ATT) $\rightarrow$ Val (GTT), Asn (ACC) $\rightarrow$ His (CAC), Phe (TTT) $\rightarrow$ Leu (TTG) and Asn (ACC) $\rightarrow$ Asp (GAC). From the analysis of the sequencing data, we were able to determine the zinc-finger-like "HHCC" motif in the amino terminus of BLV IN, that was H-$X_3$-H-$X_{25}-C-X_2$-C. It was also found the DD35E motif in the IN catalytic domain as D-$X_{56}$-D-$X_{35}$-E. It fits very well to the consensus sequences of retroviral IN as well as HHCC motif.

• BMB Reports
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• 제33권2호
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• pp.166-171
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• 2000

Thioredoxin is a multifunctional protein that is ubiquitous in microorganisms, animals and plants. Previously, the expression of the Escherichia coli thioredoxin gene (trxA) was found to be negatively regulated by cAMP. In the present study, the effect of temperature on the expression of the E. coli trxA gene was investigated. In order to examine the temperature effect, the fusion plasmid pCL70 that harbors the E. coli trxA P1P2 promoter was used. The other two fusion plasmids, pJH3 and pMH521 that were constructed in different vectors which harbor the E. coli trxA P2 promoter, were also used. When the E. coli strain MC1061/pCL70 was grown in a rich medium at $25^{\circ}C$, $34^{\circ}C$ and $42^{\circ}C$, the cells grown at $42^{\circ}C$ gave the highest $\beta$-galactosidase activity. The E. coli MC1061/pJH3 and MC1061/pMG521 cells showed increased $\beta$-galactosidase activity after the shift of the culture temperature to $42^{\circ}C$. The wild-type trxA gene of the E. coli MC1061 cells produced much higher thioredoxin activity at the higher temperature. These results support the conclusion that the E. coli trxA gene is regulated in a temperature-dependent manner. Especially the expression from its P2 promoter appeared to be sensitive to temperature.

• 인지과학
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• 제28권3호
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• pp.173-191
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• 2017

The theories of cultural evolution hold subtly or clearly different stances about definition of culture, pattern of cultural evolution, biases that affect cultural evolution, and relationship between culture and organism. However, the cultural evolution theories have a common problem to solve: As the evolutionary theory of life tries to explain the early steps and the origin of life, the cultural evolution theories also must explain the early steps of the cultural evolution and the role of the human capability that makes cultural evolution possible. Therefore, explanations of the human's unique traits including the cultural ability are related to determine which one is the most plausible among many cultural evolution theories. Theories that tried to explain human uniqueness commonly depict the coevolution of gene (organism) and culture. We will explicitly call the niche construction theory and the dual inheritance theory the 'gene-culture co-evolutionary theory'. In these theories, the most important concept is the 'concept of positive feedback'. In this paper, we distinguish between core positive feedback and marginal positive feedback, according to whether the trait that the concept of positive feedback explains is the trait of human uniqueness. Both types of positive feedback effectively explain the generality of human uniqueness and the diversity of human traits driven by cultural groups. However, this positive feedback requires an end, in contrast to negative feedback which can be continued in order to maintain homeostasis. We argue that the co-evolutionary process in the gene-culture co-evolutionary theories include only the positive feedback, not covering the cultural evolution after the positive feedback. This thesis strives to define the coevolution concept more comprehensively by suggesting the potential relationships between gene and culture after the positive feedback.

• Asian-Australasian Journal of Animal Sciences
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• 제17권6호
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• pp.743-749
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• 2004

The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

• Reproductive and Developmental Biology
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• 제37권1호
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• pp.1-8
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• 2013

In order to investigate genetic stability and gene expression profile after cloning procedure, two groups of cloned pigs were used for swine leukocyte antigen (SLA) gene nucleotide alteration and microarray analyses. Each group was consist of cloned pigs derived from same cell line (n=3 and 4, respectively). Six SLA loci were analyzed for cDNA sequences and protein translations. In total, 16 SLA alleles were identified and there were no evidence of SLA nucleotide alteration. All SLA sequences and protein translations were identical among the each pig in the same group. On the other hand, microarray assay was performed for profiling gene expression of the cloned pigs. In total, 43,603 genes were analyzed and 2,150~4,300 reliably hybridized spots on the each chip were selected for further analysis. Even though the cloned pigs in the same group had identical genetic background, 18.6~47.3% of analyzed genes were differentially expressed in between each cloned pigs. Furthermore, on gene clustering analysis, some cloned pigs showed abnormal physiological phenotypes such as inflammation, cancer or cardiomyopathy. We assumed that individual environmental adaption, sociality and rank in the pen might have induced these different phenotypes. In conclusion, the results of the present study indicate that SLA locus genes appear to be stable following SCNT. However, gene expressions and phenotypes between cloned pigs derived from the same cell line were not identical even under the same rearing conditions.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2000년도 Proceedings of 2000 KSAM International Symposium and Spring Meeting
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• pp.178-181
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• 2000

In order to develop the novel gene expression system, we introduced new control elements which could influence the foreign gene expression in animal cells. When the foreign genes are introduced into the genome of higher eukaryotic cells, the expressions from these integrated genes are often low and can vary greatly depending on the positions of the integration sites due to the complex nature of the chromatin structures (1). First we screened the various DNA sequence elements which can function as an insulator of gene expression from these position effects and can cooperate with the SV40 enhancer/promoter. Among the several DNA elements from the various sources, we identified the particular DNA element which confers the increased frequency of the positive colonies, assayed by the reporter gene from stable selections indicating significantly reduced position effects. This element also showed the several fold-increased expression level as well as the copy-number dependent expression with host cell specificity. Second we modified the transcription termination element where we introduced the specific terminator in combination with SV40 polyA signal. This modified terminator showed the increased efficiency and the level of the gene expression. By combining these two elements, we made the animal cell expression system and tested successfully for the recombinant protein productions of TGF ${\beta}$-soluble receptor, Antithrombin III, and single chain Pro-Urokinase. [Supported by grants from MOCIE]

• 미생물학회지
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• 제31권3호
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• pp.197-202
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• 1993

출아법으로 분열하는 S. cerevisiae 는 mother cell 과 bud cell 과의 연결부위근처의 원형질막 내부에 10-nm filament ring 이 존재한다. CDC3, CDC10, CDC11, CDC12 유전자가 이 filament 를 암호화할 가능성이 많은 것으로 알려져 있으며, 근래에 CDC3 와 CDC12 유전자의 생성물들이 filament ring 에 존재한다는 것이 형광현미경을 이용하여 밝혀졌으나 그 기능은 밝혀지지 않았다. 이에 본인은 10-nm filament ring 의 지능을 알아보기 위하여 ring을 이루고 있는 S. cerevisiae 의 CDC3유전자와 유사한 S. pombe 유전자의 clone 을 시도하였다. 이를 위하여 .lambda. gt11 expression vector 에 S. pombe genomic library 를 만들고 CDC3 항채를 이용하여 screen 하였다. Screen 된 유전자를 sequencing 하여 본 결과, 2개 이상의 intron 이 존재하는 것이 밝혀졌으나, 일부 부위의 아미노산 서열과 CDC3 아미노산 서열을 비교하여 본 결과, 약 62%의 유사성이 존재하였다.

• 한국양식학회지
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• 제12권3호
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• pp.175-183
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• 1999

We have shown previously that the sequence of olive flounder (Paralichthys olivaceus) hsp70-related cDAN has a high similarity with those of cognate hsc70 of other species (Kim et al., 1999; J. Aquaculture, 12:91-100). In order to investigate whether this gene encodes the congate hsc70, we examined the expression of this gene in normal and heat-shocked conditions. By in vitro translation, this gene encoded a 70 kD protein which was constitutively experessed and was not induced by heat shock. This translated protein was recognized by anti-hsp/hsc70 antibody. Tests of heat-inducibility showed that this gene was constitutively expressed in normal conditions and its expression was not increased after heat shock. The expression levels of this gene were high in stomach, gill, intestine, kidney and brain, moderate in liver, and comparatively low in overy and heart. Furthermore, Northern blot analysis of transcript expression showed that the corresponding mRNA were detected throughout embryonic development in the absence of any heat shock. These results provided evidence that olive flounder hsp70-related cDNA encoded to cognate member of hsp70 family, hsc70.