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Cloning and Structure of B - Gene from Hepatitis B Viral Genome
Rho, Hyune Mo,Kim, Seong Kee,Kim, Yong Sok,Youn, Jee Hee 한국유전학회 1986 Genes & Genomics Vol.8 No.4
The hepatitis B viral C-gene promoter region is more complex than other usual promoter region, because it contains enhancer, B-gene (X-ORF) which can encode a polypeptide 154 amino acids in length, own promoter and polyadenylation signal which follows closely C-gene initiation codon and is passed over during the first transcription run. Therefore, the elucidation of the regulation mechanism for this promoter provides a clue for the understanding of HBV life cycle. In order to study the expression and function of B-gene in relation to C-gene promoter region, we have, first, cloned a whole B-gene in SV40 expression vector. This recombinant was constructed such that the expression of B-gene (Sph I-Bgl II fragment) is under the control of SV40 early promoter and polyadenylation signal of C-gene. We have also cloned heterologous DNA fragment containing HBV enhancer, C-gene promoter and thymidine kinase gene for the purpose of studying relations between B-gene and C-gene promoter region. Another attempt to produce large amount of B-gene product in E. coli was also performed. With these recombinants and transfection experiments, we are now studying the expression of B-gene and possible functions of B-gene product in C-gene promoter region.
Characteristics of Hypervariable Regions of Mitochondrial DNA in Korean Population
Rho, Hyune Mo,Han, Jae Seok,Lee, Dong Hoon The Korea Science and Technology Center 1998 BMB Reports Vol.31 No.6
The nucleotide sequence of two hypervariable regions of the D-loop and the frequency of the 9-bp repeat in the region V of mitochondrial DNA (mtDNA) were investigated in the Korean population. Alignment of these sequences with the published reference revealed a unique pattern of base substitution and deletion compared with those of other races. The deletion and addition frequency of the 9-bp repeat in the region V was also distinct.
Expression and Regulation of HBcAg Gene in Heterologous Host System
Rho, Hyune Mo,Kim, Seong Kee,Shin, Sang Hoon 한국유전학회 1988 Genes & Genomics Vol.10 No.4
The coding sequence of HBcAg in the HBV genome (subtype adr-K) contains two inphase initiation codon: one for precore region and the other for core antigen gene. To study the expression of core antigen and the role of precore region, we subcloned the coding sequence of HBcAg gene with or without precore (pre-C) into heterologous expression vectors containing SV40 promoter, yeast PGK promoter, and lambda PL promoter. We analysed expression and secretion of core antigen in COS cells, yeast, and E. coli. To investigate the role of upstream region in the expression of the core antigen, a series of 5′ deletion mutants were constructed. In COS cells containing precore region and core antigen gene, core antigens were detected both in cell extract and cultured medium. In the case of yeast and E. coli, the antigens were detected only in cell extract in spite of the presence of precore region, suggesting that precore region could not affect the antigen-secretion in these two system.
Rho, Hyune Mo,Choi, Cheol Yong,Park, Geon Tae 생화학분자생물학회 1980 BMB Reports Vol.29 No.2
Transcription of hepatitis B viral pregenomic promoter is known to be regulated mainly by the combined interaction of enhancers I, II and the intervening regulatory sequences between the two enhancers. A positive regulatory element was identified by serial deletion and measuring the linked chloramphenicol acetyltransferase (CAT) activities, which overlapped with the 5' region of the X open reading frame. When the positive regulatory element was inserted upstream of the SV40 early promoter, it elevated SV40 promoter activity in HepG2 cells. Two cellular proteins of 110 (p110) and 33 (p33) kDa interacted with the positive element and both of them were present in the nucleus, but p110 also existed in the cytoplasm in phosphorylated form. Dephosphorylation of p110 by acid phosphatase enhanced the DNA-binding activity of p110. The p33 could bind to single-strand DNA specifically as well as to double-strand DNA.
Role of Precore Region in the Expression of Hepatitis B viral core Antigen
Rho, Hyune Mo,Kim, Seong Kee,Kim, Yong Sok 한국유전학회 1987 Genes & Genomics Vol.9 No.4
The coding sequence of HBcAg in the HBV genome contains two in-phase initiation codon, one for precore region and the other for core antigen gene. To investigate the effect of precore region on the expression of Hepatitis B viral core antigen gene, we have cloned core antigen gene with or without precore region in various expression vectors containing SV40 promoter, yeast and E. coli system. When the plasmids containing precore region were transfected to cos cells, the expression of HBcAg and that of a related antigen, HBeAg, were detected only in cell extracts in spite of the presence of precore region could not affect antigen secretion in these two systems.
Divalent Cation preference of Reverse Transcriptases with Various Synthetic Homopolymers
Rho, Hyune Mo 생화학분자생물학회 1987 BMB Reports Vol.14 No.4
Reverse transcriptases, purified from mammalian RNA tumor viruses, were tested with various template primers in the presence of different concentrations of cations. The assays are often useful in distinguishing reverse transcriptases from cellular DNA polymerases and also in identifying virus types, Results suggested that 1. Celluar DNA polymerase γ utilizes poly(A)·oligo(dT) well in the presence of Mn^(2+). 2. Type C viral reverse transcriptases utilize most synthetic homopolymers in the presence of Mn^(2+). 3. ape B and D viral reverse transcriptases utilize synthetic homopolymers in the presence of Mgz*. However, when the template primer was changed to poly(Cm)·oligo(dG), these enzymes prefered Mn^(2+) over Mg^(2+). Reversal of divalent canon preference would be an ideal category for type B and D RNA tumor viruses.