• Title/Summary/Keyword: cyt gene

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Comparison of the Genetic Relationships and Osteological Aspects in Six Branchiostegid Fish Species (Perciformes)

  • Ryu, Jung-Hwa;Kim, Jin-Koo;Park, Jung-Youn
    • Animal cells and systems
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    • v.13 no.3
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    • pp.323-329
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    • 2009
  • We analyzed partial sequences of cytochrome b (cyt-b), a mitochondrial DNA (mtDNA) gene, to determine the genetic relationships between six horsehead fish species: Branchiostegus japonicus, Branchiostegus albus, Branchiostegus auratus, Branchiostegus argentatus, Branchiostegus wardi, and an unidentified Branchiostegus species. The specimens were collected in Korea, China, Japan, and Vietnam. We compared their molecular phylogenetic relationships inferred from mtDNA cyt-b sequences with an osteological analysis. The unidentified species, B. sp., was similar to B. albus in terms of the lack of triangular silver-white dot at the posterior region of eyes (vs. large one present in B. japonicus), but was also similar to B. japonicus in terms of the presence of a straight-shaped first hemal spine (vs. a curve-shaped hemal spine in B. albus). Analysis of the mtDNA cyt-b sequences indicated that the smallest estimated sequence divergence was between the B. japonicus and B. sp. (0.70-0.94%), whereas the largest difference was between B. auratus and B. argentatus (23.06-23.36%). Both the maximum parsimony and maximum likelihood trees showed that the B. sp. was closely clustered with B. japonicus, and that B. auratus was most distant from the other species. When comparing the osteological characters, UPGMA tree showed that the B. japonicus and B. sp. were the most closely clustered species, and B. auratus was the most distantly clustered fish relative to the other species. The shape of the nasal, otolith and first hemal spine was informative for distinguishing B. auratus from the other species. These osteological differences were consistent with the differences in mtDNA.

Expression and Purification of GFPuv/Cytochrome c-552 Fusion Protein in E. coli

  • Hong, Eul-Jae;Lee, Sang-On;Choe, Jeong-U;Hong, Eok-Gi
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.550-553
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    • 2003
  • The genes of GFPuv and Cytochrome c-552 were amplified by using PCR, and then, fused each other. Fusion gene of GFPuv and Cytochrome c-552 was inserted into the pTrcHis B vector and transferred to E. coli. A fusion protein of GFPuv and Cytochrome c-552 was expressed in JM109 and BL21. This fusion protein was composed of a His-tag for the rapid one-step purification using an immobilized metal affinity chromatography.

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Amino acids at N- and C-termini are required for the efficient production and folding of a cytolytic γ-endotoxin from Bacillus thuringiensis

  • Thammachat, Siriya;Pathaichindachote, Wanwarang;Krittanai, Chartchai;Promdonkoy, Boonhiang
    • BMB Reports
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    • v.41 no.11
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    • pp.820-825
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    • 2008
  • Bacillus thuringiensis Cyt2Aa toxin is a mosquito-larvicidal and cytolytic $\delta$-endotoxin, which is synthesized as a protoxin and forms crystalline inclusions within the cell. These inclusions are solubilized under alkaline conditions and are activated by proteases within the larval gut. In order to assess the functions of the N-and C-terminal regions of the protoxin, several N- and C-terminal truncated forms of Cyt2Aa were constructed. It was determined that amino acid removal at the N-terminal, which disrupts the $\beta$1 structure, might critically influence toxin production and inclusion formation. The deletion of 22 amino acids from the C-terminus reduced the production and solubility of the toxin. However, the removal of more than 22 amino acids from the C-terminus or the addition of a bulky group to this region could result in the inability of the protein to adopt the proper folding. These findings directly demonstrated the critical roles of N- and C-terminal amino acids on the production and folding of the B. thuringiensis cytolytic $\delta$-endotoxin.

A Phylogenetic Study of Korean Rodents (Muridae, Sciuridae) Based on Mitochondrial and Nuclear DNA

  • Jung, Gi-La;Lee, Seo-Jin;Kim, Chuel-Kyu;Lee, Hang;Kim, Chang-Bae
    • Animal Systematics, Evolution and Diversity
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    • v.26 no.2
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    • pp.99-104
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    • 2010
  • The subfamily Murinae is a very controversial group concerning their phylogenetic relationship. Previous studies could not resolve phylogeny among four genera Apodemus, Micromys, Mus and Rattus of the Muridae. In the present study, eight rodent species resident in South Korea were collected and phylogenetically analyzed based on sequence data of five mitochondrial and nuclear DNA regions: 12S rRNA, cytochrome b gene (cyt b), cytochrome oxidase II (COII), control region of mitochondrial DNA, and a thyroglobulin (Tg) of nuclear DNA. According to the phylogeny of the concatenated data, M. musculus separated early in Murinae (ML 100%; BA 1.00 pp) and the genus Rattus grouped with the harvest mouse, M. minutes; these were separated from the genus Apodemus with relatively strong support (ML 74%; BA 0.76 pp). The Siberian chipmunk population was also examined using the five genes to obtain better resolution. The phylogeny for Korean rodents determined using the 12S rRNA, cyt b, COII and control regions discriminated the Siberian chipmunk populations from Korea, Russia, and China.

Introduction of Stay Green Mutant for the Development of Black Seed Coat and Green Cotyledon Soybean Variety (녹색자엽 검정콩 품종 육성을 위한 Stay green 변이체 활용)

  • Kang, Sung-Taeg;Seo, Min-Jung;Moon, Jung-Kyeong;Yun, Hong-Tae;Lee, Young-Ho;Kim, Si-Ju;Hwang, Young-Sun;Lee, Suk-Ki;Choung, Myoung-Gun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.3
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    • pp.187-194
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    • 2010
  • The soybean stay green mutant genotype (SSG) derived from the nuclear gene, d1d2, and cytoplasmic gene, cytG, inhibit the breakdown of chloroplast in the leaves, pod walls, seed coats, and embryos during maturity. Soybean seed with black seed coat and green cotyledon (SBG) are preferred than black seed coat with yellow cotyledon (SBY) especially for cooking with rice and as source of traditional food in Korea. The researchers evaluated the seed's chlorophyll content of SSG and introduced SSG to the SBG variety breeding program. The seed chlorophyll content of SSG with d1d2 was $39.93{\sim}60.80\;{\mu}g/g$ and SSG with cytG $38.08{\sim}39.89\;{\mu}g/g$. The Korean SBG variety which was derived from SSG with cytG, contains $16.35{\sim}37.73\;{\mu}g/g$. The composition of seed chlorophyll differs according to the genetic background of SSG genotype. Inheritance study showed that cotyledon color was segregated 15:1 (yellow:green) at $F_2$ seed indicating two recessive genes control green cotyledon as revealed by previous study. Only less than 3% soybean lines showed black seed coat with green cotyledon among crosses SBY and SSG (d1d2). Results showed that SSG with d1d2 can be used as a good source for SBG with high chlorophyll content in the seed cotyledon, but due to the complex genetic behavior, breeding resource of SBG with d1d2 should be prepared to improve the breeding efficiency for development SBG variety.

Prohibitin Induces Apoptosis in BGC823 Gastric Cancer Cells Through the Mitochondrial Pathway

  • Zhang, Long;Ji, Qing;Ni, Zhen-Hua;Sun, Jian
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3803-3807
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    • 2012
  • Prohibitin (PHB), an evolutionarily-conserved protein, has been found to be over-expressed in gastric cancer and be closely related with tumor malignancy. In this study, to investigate the relationship between PHB expression and cell apoptosis in the BGC823 gastric cancer cell line, low and high expression PHB in BGC823 cells was accomplished using RNA interference technology and gene transfer techniques. Cell proliferation, cell cycling, apoptosis, Bax, Bcl-2 and Cyt.c protein expression and the activation of Caspase-3,9 were assessed after 48h. Over-expression of PHB gene in BGC823 cells resulted in slow cell growth, cell arrest in G2 phase, and an increased apoptosis ratio while the opposite was found for PHB under-expressing cells. In PHB over-expressing cells, the expression of Bax gene was increased, the expression of Bcl-2 was decreased, the activation level of Caspase-3, 9 was increased, but the activation level of Caspase-8 demonstrated no change. These results indicate that PHB induced apoptosis through the mitochondrial pathway.

Geographic Variation of Granulilittorina exigua (Littorinidae, Gastropoda) in Korea Based on the Mitochondrial Cytochrome b Gene Sequence

  • Song, Jun-Im;Suh, Jae-Hwa;Kim, Sook-Jung
    • Animal cells and systems
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    • v.4 no.3
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    • pp.267-272
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    • 2000
  • Partial sequence of the mitochondrial cytochrome b gene was analyzed to investigate genetic variation from 10 geographic populations of Granulilittorina exigua in Korea. The sequence of 282 base pairs was determined by PCR-directed silver sequencing method. The sequences of two species within the genus Littorina reserved in NIH blast search were utilized to determine geographic variations of species referred. The levels of mtDNA sequence differences were 0.00-2.54% within populations and 0.71-4.43% between populations. There were four amino acid differences between representative species of the genera Granulilittorina and Littorina, but no differences within populations of the genus Granulilittorina. The UPGMA and the N-J trees based on Tamura-Nei genetic distance matrix were constructed, which showed that the genus Granulilittorina was divided into three groups such as eastern (even exception for Tokdo population), southern, and western regional populations. The degrees of genetic divergence within populations of each group were p=0.021, p=0.019, and p=0.018, respectively. The divergence between the eastern and southern populations was p=0.032, showing closer relationship than with the western populations (p=0.052). Based on the diverged time estimation, the eastern and southern populations of Granulilittorina exigua in Korea diverged from the western populations about 2.1 MYBP, and the eastern and southern populations diverged from each other about 1.3 MYBP.

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Characteristics of Hemolysin in Mosquitocidal Bacillus thuringiensis strain 21-2 (모기 살충성 Bacillus thuringiensis 21-2균주의 용혈성 내독소 단백질의 특성)

  • 김광현;김위종;김영희;김병우
    • Microbiology and Biotechnology Letters
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    • v.30 no.3
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    • pp.230-234
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    • 2002
  • To describe characteristics of a hemolysin in mosquitocidal Bacillus thuringiensis subsp. gyangiensis strain 21-2, Escherichia coli HB101 was transformed with a gene encoding hemolysin in the strain 21-2. Transformant 47 con-tained 2.5 kb DNA was selected by ELISA, immunoblot and DNA electrophoresis. Transformant 47-5 was recon-structed after digestion of the 2.5 kb DNA with Hind m. Transformant 47-5 contained 1.8 kb DNA and expressed 23 kDa Protein which had mosquitocidal activity to Aedes aegypti. The 23 kDa Protein itself in vitro didn't show hemolytic activity on human erythrocytes, but the protein had the activity after proteinase K treatment.

Genetic Diversity among Local Populations of the Gold-spotted Pond Frog, Rana plancyi chosenica (Amphibia: Ranidae), Assessed by Mitochondrial Cytochrome b Gene and Control Region Sequences

  • Min, Mi-Sook;Park, Sun-Kyung;Che, Jing;Park, Dae-Sik;Lee, Hang
    • Animal Systematics, Evolution and Diversity
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    • v.24 no.1
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    • pp.25-32
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    • 2008
  • The Gold-spotted pond frog, Rana plancyi chosenica, designated as a vulnerable species by IUCN Red list. This species is a typical example facing local population threats and extinction due to human activities in South Korea. A strategic conservation plan for this endangered species is urgently needed. In order to provide information for future conservation planning, accurate information on the genetic diversity and taxonomic status is needed for the establishment of conservation units for this species. In this study, we used a molecular genetic approach using the mitochondrial cytochrome b gene and control region sequences to find the genetic diversity of gold-spotted pond frogs within South Korea. We sequenced the mitochondrial DNA cytochrome b gene and control region of 77 individuals from 11 populations in South Korea, and one from Chongqing, China. A total of 15 cytochrome b gene haplotypes and 34 control region haplotypes were identified from Korean gold-spotted pond frogs. Mean sequence diversity among Korean gold-spotted pond frogs was 0.31% (0.0-0.8%) and 0.51% (0.0-1.0%), respectively. Most Korean populations had at least one unique haplotype for each locus. The Taean, Ansan and Cheongwon populations had no haplotypes shared with other populations. There was a sequence divergence between Korean and Chinese gold-spotted pond frogs (1.3% for cyt b; 2.9% for control region). Analysis of genetic distances and phylogenetic trees based on both cytochrome b and control region sequences indicate that the Korean gold-spotted pond frog are genetically differentiated from those in China.

Qualitative PCR Detection of Stack Gene GM Rice (LS28 X Cry1Ac) Developed in Korea (국내개발 stack gene GM 벼(LS28 X Cry1Ac)에 대한 정성 PCR 분석)

  • Shin, Kong-Sik;Park, Jong-Hyun;Lee, Jin-Hyoung;Lee, Si-Myung;Woo, Hee-Jong;Lim, Sun-Hyung;Kim, Hae-Yeong;Suh, Seok-Cheol;Kweon, Soon-Jong
    • Journal of Applied Biological Chemistry
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    • v.52 no.1
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    • pp.1-7
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    • 2009
  • For the development of qualitative PCR detection method of genetically modified (CM) rice, rice species-specific gene, OsCc-1 (rice cytochrome c gene), was selected as suitable far use as an endogenous gene in rice. The primer pair OsCytC-5'/3'with 111 bp amplicon was used for PCR amplification of the rice endogenous gene, OsCc-1 and no amplified product was observed from 8 different crops as templates. Qualitative PCR method was carried out with stack traits of L528$\times$CryIAc1 GM rice developed in Korea. For the qualitative PCRs, some primer pairs were designed with a construct-specific and event-specific type based on T-DNA and junction sequences of T-DNA in GM rice. Actck-5'/3' amplifying between actin promoter and OsCK1 gene introduced in LS28 gave rise to an amplicon 306 bp; also, CrLB-5'/3' from CryIAcl and CKRB-5'/3'amplifying the junction region of T-DNA and genome sequence from LS28 as event-specific primers gave rise to an amplicon 142 bp and 91 bp, respectively. These primer pairs for the detection of event-specific targets not produced PCR amplicons on non-CM rice and various crops in contrast to event lines. Therefore, in this study we verified that event-specific primers were effective to specifically detect stack trait lines and demonstrated that this method presented could be provided with the detection-method data for risk assessment analysis of GM rice to be commercialized.