• 대한바이러스학회지
• /
• 제29권4호
• /
• pp.261-269
• /
• 1999

The relationship between second messenger cGMP and human cytomegalovirus (HCMV) replication was investigated. First, the intracellular level of cGMP ([cGMP]i) in HCMV-infected cells was measured. The [cGMP]i increased at early times after HCMV infection, reached maximum level at 12 hr and returned to basal level at 24 hr after virus infection, while [cGMP]i in mock-infected cells remained relatively unchanged. Increasing [cGMP]i resulted in enhanced transcription of HCMV major immediate early gene. For early gene expression, cGMP had varying effect. Expression of 1.2 kb RNA decreased and 2.2 kb RNA increased with increasing cGMP, while 2.7 kb RNA gene expression was not affected. HCMV early genes are regulated by immediate early gene, and the effect of cGMP on the regulatory effect of major immediate early gene on early genes was investigated. In the absence of cGMP, major immediate early gene repressed 2.7 kb RNA gene expression, while 1.2 kb RNA and 2.2 kb RNA early genes were not significantly affected. In the presence of $1\;{\mu}M$ cGMP, however, major immediate early gene stimulated the expression of three early genes.

• 환경생물
• /
• 제23권3호
• /
• pp.269-274
• /
• 2005

The 2D/E gel analysis for polypeptide expression reflecting I-18 C gene (early-ecdysterone inducible gene) has conducted the emerged C. riparius adults from larval phase exposure to tebufenozide acting as an ecdysteroidal molting hormone. Control group, the amount of ORE II of the I-18 C gene was larger than that of ORE I of this gene. After treatments, ORE I of the I-18 C gene was overexpressed as the polypeptide, whereas ORF II of this gene was expressed as the polypeptide and was clearly reduced expression. Accordingly, we consider that tebufenozide exhibited endocrine disruptions related processing of ecdysteroid receptor protein reflecting ORF II of I-18 C gene. Also, earlier emergence day was related overexpressed polypeptide reflecting ORE I of I-18 C gene. In this study result, tebufenozide induced changing of physiological condition, and then polypeptide expression reflecting early-ecdysterone inducible I-18 C gene was different between control group and exposure group.

• Journal of Microbiology
• /
• 제38권1호
• /
• pp.24-30
• /
• 2000

US3 gene is a member of the human cytomegalovirus (HCMV) immediate early gene. Although the precise functions of the US3 gene in HCMV replication and pathogenesis are not known, it has been reported to play a role in inhibiting major histocompatibility class I antigen presentation. For further knowledge of US3 gene expression, rabbit polyclonal antiserum of the US3 gene product was used for indirect immunofluorescence assay. In permissive human foreskin fibroblast (HFF) cells, US3 gene expression was detectable as crescent or half-moon shape in the perinuclear region at immediate early times after virus infection. HFF cells infected with mutant HCMV lacking US3 open reading frames were negative for US3 immunofluorescence assay. Double immunofluorescence assay using monoclonal antibody to gamma adaptin (specific for the Golgi complex) and rabbit anti-US3 antiserum revealed that US3 gene product could be localized to the Golgi complex. At later time after HCMV infection, US3 gene products were detected as globular aggregates in the cytosol. These aggregates were positive for gamma adaptin and stained with preimmune serum, suggesting a nonspecific reaction to the Golgi complex. Northern blot analysis revealed that transcription of US3 was observed only during immediate early times after virus infection (until 6 h postinfection). Therefore US3 gene expression appears to be confined to immediate early time and its gene products are localized to the Golgi complex as crescent shaped forms in the perinuclear cytoplasm.

• Asian-Australasian Journal of Animal Sciences
• /
• 제8권5호
• /
• pp.449-454
• /
• 1995

The present study was conducted to assess gene expression of bacterial lacZ driven by the SV40 promoter at early developmental stages of bovine embryos. The lacZ gene was linearized with BamHI digestion and introduced into the pronucleus by microinjection at 20 hrs after the commencement of in vitro fertilization. Intact bovine blastocysts were not stained with X-Gal, suggesting that there is no endogenous beta-galactosidase activity in these blastocysts. In contrast, the bovine blastocyst cells microinjected with the lacZ gene exerted a characteristic greenish-blue color originating from the bacterial beta-galactosidase activity, albeit at a low rate, i.e. 2.1% of the total fertilized oocytes injected. It was concluded, therefore, that the lacZ gene driven by the SV40 promoter could be used for an indirect screening method in which the presence of transgene is evaluated from the product of transgene expression.

• BMB Reports
• /
• 제32권5호
• /
• pp.456-460
• /
• 1999

The promoter of the HSV-1 VP16 gene contains binding sites for the cellular transcription factors such as USF, CTF, and Sp1, each of which affects basal level expression of the VP16 gene. Transcription of the VP16 gene was induced by viral immediate-early proteins, ICP0 and ICP4, in a synergistic manner but repressed by ICP22. To gain further insight into the role of ICP0 in the expression of the VP16 gene during virus infection, several mutants with deletions in each of their transcriptional regulatory elements were generated. According to transient gene expression assays of these mutants using the CAT gene as a reporter, the USF and CTF binding sites were necessary for efficient induction of the promoter in the presence of transfected ICP0 or during virus infection, whereas the Sp1 binding site had little effect on ICP0-mediated VP16 expression. These results indicate that the immediate early proteins of HSV-1 regulate expression of the VP16 gene during virus infection by modulating the activities of cellular transcription factors such as USF and CTF.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제13권1호
• /
• pp.23-30
• /
• 2006

BmCu/Zn SOD was isolated from early embryo of Bombyx mori using microarray analysis. The BmCu/Zn SOD gene was observed during the early embryonic stage with the strongest signal found at the unfertilizaion, fertilization and blastoderm stages. The BmCu/Zn SOD gene encodes a protein of 154 amino acids with a calculated Mr of 15 kDa. The deduced amino acid sequence of BmCu/Zn SOD indicated that the residues that form on the Cu/Zn binding site are conserved and that the sequence is a 60% identity to that of M. domestica. In a phylogenetic tree, Bm SOD was also close to Drosophila SODs rather than other insect SODs. The BmCu/Zn SOD gene exists as a single copy in the genome. Transcripts of BmCu/Zn SOD cDNA were identified by northern blot analysis. The expression of the BmCu/Zn SOD gene was observed weakly in most of larvae, pre-pupae, pupae and adult tissues. Also, the BmCu/Zn SOD gene was observed in early embryonic stage. Although the roles of SODs remains to be further elucidated, the high expression of BmCu/Zn SOD gene at before 24 h post fertilization suggests that this gene is of general importance during early embryogenesis in the Bombyx mod.

• BMB Reports
• /
• 제29권5호
• /
• pp.474-480
• /
• 1996

We present a study of evolutionary changes in expression of actin genes among closely related sea urchin species that exhibit different modes of early development. For this purpose, polyclonal antisera raised against peptides from the carboxyl terminus of the HeCyI cytoskeletal actin of Heliocidaris erythrogramma were used. H. erythrogramma is a direct developing sea urchin that proceeds from embryonic to adult stages without an intervening feeding larval stage. Expression patterns of the CyI actin isoform were compared with those of Heliocidaris tuberculata and to a related sea urchin Strongylocentrotus purpuratus, which both produce a feeding pluteus larval stage. The CyI actin of all three species is expressed in the same cell types. However, its expression patterns have been changed with reorganization of early cell lineage differentiation, which is apparent among the three species. Thus. evolutionary changes in CyI actin gene expression patterns are correlated with not only phylogenetic relationship, but developmental mode. The implication of this observation is that evolutionary changes in expression patterns of histospecific genes may underlie the emergence of novel developmental processes.

• 한국미생물·생명공학회지
• /
• 제29권1호
• /
• pp.18-24
• /
• 2001

누에 핵다각체병 바이러스의 gp64 프로모터를 이용한 transient 발현 벡터를 제작하기 위해서 gp64 유전자의 구조를 분석하였다. Southern blotting 분석을 통해 genome DNA에서 gp64 유전자를 탐색하기 gp64 구조유전자를 포함하는 2,277 nucleotide의 염기를 분석하였으며 gp64의 early, late 프로모터 발현을 조절하는 인자들을 확인하였다. gp64프로모터를 이용한 transient 발현 벡터를 제작하고 외래유전자로써 lacZ 유전자를 Bm5 세포주에서 transient 발현시켰다. 세포주 내에 도입된 플라스미드 DNA의 안정성을 확인하였으며, gp64 프로모터의 외래유전자 발현성 여부를 조사하기 위하여 gp64 프로모터 하에 laxZ 유전자를 가지는 재조합 바이러스를 제작하고 $\beta$-galactosidase in 냐셔 staining을 수행한 결과 전체적인 발현량은 매우 약한 것으로 판단되었다. BmNPV-K1의 gp64 프로모터를 이용한 벡터는 더욱 민감한 표지 유전자를 발현시켜 재조합 바이러스의 분리에 이용하거나 숙주세포에 독성을 보이는 유전자 산물의 소량 발현에 더욱 유용할 것으로 판단된다.

• Journal of Animal Science and Technology
• /
• 제48권3호
• /
• pp.345-352
• /
• 2006

지방합성이 왕성하게 진행되는 비육후기의 근육내 지방합성에 있어서, 지방산 및 당의 이용경로 및 이에 따른 근육내 지방합성과정을 규명하기 위해서 지방산 및 당 운반 유전자인 FABP4, GLUT4와 근육내 지방대사 주요유전자인 ACL, ACC, LPL 및 SCD 유전자의 mRNA 발현양상을 분석하였다. 한우 비육전기 및 후기 각 3두를 공시하여 total RNA 추출 및 1st cDNA 합성하여 SYBR green을 이용하여 real-time PCR 분석을 각 유전자별로 3반복씩 수행하였다. FABP4 유전자는 비육전기에 비해 비육후기에 있어서, 3배 이상 유전자 발현이 증가함을 확인할 수 있었고, GLUT4 유전자는 비육전기 및 비육후기에서 큰 차이를 보이지 않았다. 또한 근육내 지방합성 주요유전자인 ACL, ACC, LPL 및 SCD 유전자의 발현량은 비육후기에서 ACL 유전자가 3.8배, ACC 유전자는 2.7배, LPL 유전자는 3.5배, 그리고 SCD 유전자는 7.5배 발현량이 증가하였다. 따라서 본 연구를 통하여 한우의 비육후기에서 근육내 지방합성은 FABP4에 의한 지방산 유입의 증가와 더불어 ACL 유전자에 의해서 지방산 합성의 중간대사물인 acetyl-CoA가 합성되고, 이 중간 대사물을 이용하여 ACC 및 SCD 유전자에 의해서 긴 사슬 지방산 합성이 왕성하게 일어남을 알 수 있었다.

• 한국양식학회지
• /
• 제12권4호
• /
• pp.275-281
• /
• 1999

The utility of RSV-LTR and carp beta-actin promoters was evaluated in a marine flatfish species, Limanda Yokohamae by examining successful expression of transgenic DNA in muscles (transfected by direct injection) and in early embryos (transformed by lipofected sperm). The expressed pattern of injected DNA in skeletal muscles was dependent on the DNA amount injected. The activity reached to maximal level at 48 hours post injection, and persisted up ot 1 month transiently. Gene transfer into early embryo of this species was successfully achieved using lipofected sperm with the efficiency ranging 36.8% to 48.1%. The expression of transgene during embryonic development was shown as stage-specific and transient.