• Journal of Microbiology
• /
• 제41권2호
• /
• pp.129-136
• /
• 2003

The Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMPI) exhibits considerable sequence heterogeneity among EBV isolates. Seven distinct EBV strains have been defined based on sequence polymorphisms in the LMPI gene, which are designated China 1, China 2, China 3, Alaskan, Mediterranean, NC, and the B95-8 strains. In this study, we analyzed a 30-bp deletion and sequence variations in the carboxy-terminal region of the LMPl gene in 12 EBV isolates from spontaneous lym-phoblastoid cell lines derived from individuals with non-EBV associated cancers in Korea. Eleven of the 12 isolates showed a 30-bp deletion spanning LMPI amino acids 342 to 353, suggesting a high prevalence of the LMPI 30-bp deletion variant among EBV isolates in Korea. In addition, all 12 isolates had a 15-bp common deletion in the 33-bp repeat region and multiple base-pair changes relative to the prototype B95-8 EBV strain along with variations in the number of the 33-bp repeats. The bp changes at positions 168746, 168694, 168687, 168395, 168357, 168355, 168631, 168320, 168308, 168295, and 168225 were highly conserved among the isolates. Comparative analysis of sequence change patterns in the LMPI carboxy-terminal coding region identified nine 30-bp deletion variants as China 1, two deletion variants as a possible interstrain between the Alaskan and China 1 strains, and a single undeleted variant as a possible variant of the Alaskan strain. These results suggest the predominance of the China 1 EBV strain in the Korean population.

• BMB Reports
• /
• 제29권5호
• /
• pp.468-473
• /
• 1996

The P38 promoter of minute virus of mice (MVM) is a very weak promoter which is strongly transactivated by viral nonstructural proteins. To analyze the upstream sequence of the P38 promoter which is responsible for the transactivation by nonstructural proteins in MVM, chloramphenicol acetyltransferase (CAT) reporter plasm ids containing a series of 5' deletion and internal deletion mutants of the P38 promoter were constructed. The wild type and mutant CAT constructs of P38 promoter were cotransfected into murine A92L fibroblast cells with a plasmid expressing viral nonstructural proteins by DEAE-dextran method. Each promoter activity was analyzed by CAT assay. As previously reported (Ahn et al., 1992), the proximal DNA cis-elements required for transactivation of the MVM P38 promoter are GC box and TATA box. However, the analysis of 5' deletion mutants showed that H-l tar like sequence (MVM TAR) which is located between -143 and -122 relative to the transcription initiation site is also required for transactivation of the P38 promoter by nonstructural proteins. Interestingly, even if the MVM TAR was removed by internal deletion, the level of the transactivation is still 70% of wild type level of transactivation. We also found that, in addition to the MVM TAR motif, there are two other motifs which are similar to the MVM TAR sequence. When these TAR like motifs were further deleted, the levels of transactivation were decreased further. Taken together, the MVM TAR sequence and TAR like motifs located upstream of P38 promoter are playing an important role for the transactivation of P38 promoter by nonstructural proteins in minute virus of mice.

• Reproductive and Developmental Biology
• /
• 제30권4호
• /
• pp.293-299
• /
• 2006

In order to generate transgenic goats expressing human growth hormone (hGH) in their mammary glands, goat ${\beta}-Casein/hGH$ hybrid gene was introduced into goat zygotes by pronuclear microinjection. DNA-injected embryos were transferred to the oviduct of recipients at 2-cell stage or to the uterus at morula/blastocyst stage after cultivation in glutathione-supplemented mSOF medium in vitro. Pregnancy and survival rate were not significantly different between 2-cell embryos and morula/blastocysts transferred to oviduct and uterus, respectively. One transgenic female goat was generated from 153 embryos survived from DNA injection. Southern blot analysis revealed that the transgenic goat harbored single-copy transgene with a partial deletion in its sequences. Despite of the partial sequence deletion, the transgene was successfully expressed hGH at the level of $72.1{\pm}15.1{\mu}g/ml$ in milk throughout lactation period, suggesting that the sequence deletion had occurred in non-essential part of the transgene for the transgene expression. Unfortunately, however, the transgene was not transmitted to her offspring during three successive breeding seasons. These results demonstrated that goat ${\beta}-casein/hGH$ gene was integrated into the transgenic goat genome in a mosaic fashion with a partial sequence deletion, which could result in a low level expression of hGH and a failure of transgene transmission.

• BMB Reports
• /
• 제30권1호
• /
• pp.41-45
• /
• 1997

The present study was performed to analyze the molecular mechanism which dictates the transcription regulation of the $O^6$-methylguanine-DNA methyltransferase (MGMT) gene in Saccharomyces cerevisiae. Previously we identified one possible upstream repressing sequence (URS) in MGMT promoter by promoter deletion and competition analysis. In this paper we report another regulatory element (UAS: upstream activating sequence. -213 to -136) which affects the transcription activity of MGMT promoter. Gel mobility shift assay and Southwestern blot analysis using UAS probe showed several specific proteins which were able to bind to this sequence.

• Applied Biological Chemistry
• /
• 제38권5호
• /
• pp.387-392
• /
• 1995

대두 glycinin 유전자의 조직 특이적이고 분화 발달 특이적인 발현 조절 메카니즘을 연구하기 위하여 Gy2 유전자의 5' upstream 부위 염기서열을 조사한 결과, glycinin 유전자의 발현을 조절하는 인자로 여겨지는 여러 가지 조절 인자들을 발견하였다. 진핵세포 유전자에 공통적으로 존재하는 TATA box와 AGGA box가 존재하고, 종자 저장 단백질에서 공통적으로 발견되는 embryo factor binding sequence, RY repeat CACA sequence, ${\beta}$-conglycinin enhancer 와 유사한 sequence 등이 발견되었다. 이러한 조절 요소들이 Gy2 유전자의 발현 조절에 미치는 영향을 알아보기 위해 Gy2유전자의 5' upstream부위를 Exo III nuclease와 여러가지 제한효소를 이용하여 일련의 deletion mutants를 제조한 후 GUS 유전자와 결합시켰다. 이들 여러가지 chimeric constructs를 대두 원형질체에 전입하고 원형질체로부터 추출물을 분리하여 GUS 활성을 조사한 결과, $-28l{\sim}-223$ 혹은 $-l70{\sim}-122$ 부위를 포함하였을 경우 활성이 감소하였고, $-223{\sim}-170$ 혹은 $-l22{\sim}-16$ 부위를 포함하였을 경우 활성이 높게 나타났다. 이러한 Gy2 유전자의 이중적인 발현 양상은 glycinin 유전자의 발현조절에 음성 조절 요소와 양성 조절 요소가 관여하고있다는 사실을 제시해 주고 있다. 또한 이들 여러가지 chimeric constructs로 형질 전환된 담배의 종자와 잎에서 GUS활성을 조사한 결과, CaMV promoter를 포함하는 chimeric construct는 종자와 잎에서 모두 활성을 나타냈으나, Gy2 Promoter를 포함하는 chimeric constructs는 종자에서만 GUS 활성을 나타내고 잎에서는 활성이 나타나지 않는 조직 특이적인 발현 양상을 나타내었다.

• 미생물학회지
• /
• 제30권2호
• /
• pp.141-148
• /
• 1992

In order to overexpress bacteriophage PRD1 DNA polymerase in E. coli cells, the 2 kb HaeII fragment was isolated from phage genomic DNA. This fragment was then cloned into pEMBL/sup ex/ 3-expression vector. A specific 57bp deletion was performed by using uracil containing ss DNA and oligonucleotide spanning each region to remove an unwanted non-coding region. After this deletion, the PRD1 DNA polymerase gene is totally under the control of the vector promoter and SD sequence. Upon heat induction, a protein with an apparent size of 68 kdal was overexpressed as an active PRD1 DNA polymerase. The expression of PRD1 DNA polymerase was about 1% of total E. coli protein.

• 미생물과산업
• /
• 제14권1호
• /
• pp.12-16
• /
• 1988

Allantoin 분해 유전자들중 highly inducible 한 DAL7, DUR1,2및 constitutive한 DAL5 gene의 promoter를 deletion 방법에 의해 발현에 필요한 최소 DNA seqyence 부위를 정한후 이 DNA seqyence를 다시 oligonucleotide 합성방법에 의해 합성하여 Cyc 1-LacZ expression vector에 삽입하여 효모내에서 LacZ의 발현이 삽입한 DNA sequence에 의해 영향을 받는 정도를 측정하여 (.betha.-galactosidase activity) deletion 방법에 의해 결정한 이 DNA dequence들이 직접 발현유도에 관여하는가를 조사하였다.

• Journal of Plant Biology
• /
• 제33권4호
• /
• pp.303-307
• /
• 1990

A genomic clone carrying a proteinase inhibitor I sequence was isolated and characterized. The clone contained a 0.7 kb EcoRI fragment hybridized with tomato inhibitor I cDNA. The nucleotide sequence of the EcoRI fragment revealed presence of a truncated form of a proteinase inhibitor I gene of potato. The truncated gene contained the 5' flanking region and the first exon of a functional proteinase inhibitor I gene. Although the 5' flanking region contained the regulatory sequences TATAAA and CCACT, a deletion of 40 bp occurred between them.

• 한국식물병리학회지
• /
• 제14권6호
• /
• pp.612-613
• /
• 1998

The nucleotide sequence of hop stunt viroid HHSVd) Kh strain was sequenced by the reverse transcription and polymerase chain reaction. It consists of 296 nucleotides, and differs by one nucleotide deletion of cytosine at the position of 295 from the HSVd-K strain which consists of 297 nucletoides.

• Animal cells and systems
• /
• 제2권2호
• /
• pp.259-267
• /
• 1998

A deletion of one out of the two copies of 9-bp repeat sequence (CCCCCTCTA), between the cytochrome oxidase II and Iysine tranfer RNA (COII/$tRNA^{Lys}$) genes in human mitochondrial DNA (mtDNA) has been used as a polymorphic anthropological marker for people of east Asian origin, and to lesser extent, Pacific and African populations. We searched for the 9-bp deletion of the intergenic COII/$tRNA^{Lys}$ Lys region in two Korean populations (175 from Seoul and 38 from Cheju) and examine the distibution of this deletion in world populations. The 9-bp deletion was detected directly by electrophoresis of the polymerase chain reaction (PCR)-amplified nucleotide(nt) 8211-8310 mtDNA fragment. The frequencies of the 9-bp deletion were significantly different between the Seoul (16%) and Cheju (8%) populations. Examination of data from the world populations suggests a geographic gradient. The frequency reaches its highest values in some Pacific island populations and decreases along the southeast Asia-Siberia transect. In spite of this geographic gradient, Mongoloid populations including Korean, Chinese, Japanese, and Mongolian populations were relatively homo-geneous with regard to the 9-bp deletion type of the intergenic COII/$tRNA^{Lys}$ region. These results indicate Koreans are genetically related to northeast Asian populations, and have a maternal mongoloid ancestry. Therefore, the 9-bp deletion of the intergenic COII/$tRNA^{Lys}$ region will provide significant information to elucidate the historical patterns of migration of the Mongoloids.