• 제목/요약/키워드: KB-C2

검색결과 471건 처리시간 0.03초

구기자나무(Lycium chinense Mill.)로의 rolC유전자 도입에 미치는 요인 (Factors Affecting Introduction of rolC Gene in Lycium chinense Mill.)

  • 박용구;최명석;김병원;정원일;노광수
    • 식물조직배양학회지
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    • 제22권6호
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    • pp.329-334
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    • 1995
  • 효과적인 형질전환 시스템을 이용하여 왜화유전자인 rolC 유전자를 구기자나무로의 형질전환 시스템을 확립하였다. 침으로 자극된 엽절편을 2.0 mg/L zeatin이 함유된 3/2 MS배지에 배양하였을 때 엽표면으로부터 줄기재분화가 되었다. 그러나 여러 농도의 kanamycin sulfate와 2.0 mg/L Aeatin이 함유된 배지에 엽절편을 배양하였을 때는 kanamycin sulfate의 농도가 증가할수록 줄기 유발수가 감소하는 것을 볼 수 있었으며, 적정선발농도는 10 mg/L이었다. 엽절편은 공동배양 시간에 따라서 생존율과 줄기재분화에 매우 큰 영향을 미쳤다. 엽절편의 생존율은 dipping할 경우가 가장 좋았으며, 배양시간이 길수록 엽절편의 백색화가 관찰되었고, 생존율이 급격히 감소하는 것을 볼 수 있었다. 줄기재분확에 가장 적합한 공동배양 시간은 24시간으로 나타났다. 공시균주와 24시간 동안 공동배양한 엽절편을 10mg/L의 kanamycin sulfate와 2.0 mg/L zeatin이 함유된 줄기 유도배지에 배양한 결과, 105개의 엽절편 중 80개의 엽절편이 생존하였으며, 그 중에서 15개의 재분화된 줄기를 얻었다. 재분화된 줄기들은 형질전환여부를 판명하기 위해 1차적으로 10 mg/L kanamycin sulfate가 함유된 배지에 옮겨 4 주간 배양한 결과, 항생제에 대해서 저항성을 가진 5개의 식물체를 선발할 수 있었다. 2차적으로 rolC유전자와 NPTII 유전자 도입의 유무를 검증하기 위하여 Southern 분석을 행한 결과, 구기자의 형질전환 식물체에서 rolC 유전자 probe의 coding sequence와 동일한 것으로 생각되는 1kb위치와 NPTII probe의 coding sequence와 동일한 것으로 생각되는 2.6kb 위치에서 각각의 밴드를 확인할 수 있었다.

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Archicitrus와 Metacitrus로부터 Mitochondrial DNA의 분리 및 특성 (Isolation and Characterization of Mitochondrial DNA from Arehicityars and Metacitrus)

  • 이숙영;박민희
    • 한국식품영양학회지
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    • 제8권4호
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    • pp.307-317
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    • 1995
  • The purity of mtDNAs isolated from Archicitrus and Metacitrus leaves was higher in percoll density gradient centrifugation than differential and sucrose density gradient centrifugation. The most clear mtDNAs were obtained from mitochondria included in the Interface band of between 21% and 45% under isomotic, low viscosity conditions in the three step discontinuous percoll density gradient centrifugation. DNase treatment to the crude mitochondrlal suspension still more increased purity of mtDNA by the effective removal of the nuclear and chloroplast DNA and mtDNAs were appeared as a single band at middle position of tube by EtBr /cscl density gradient centrifugation. Agarose gel electrophoresis of mtDNAs resolved a single, broad band containing high molecular weight DNAs In all preparation. Yield of mtDNAs was about 110 and 2 ug Per 2009 in mature and immature leaves respectively. The mtDNA fragment patterns showed by EcoR I treatment were indistinguishable with respect to nom bet and position of bands in Archicitrus and Metacitrus. In the pattern of Hind E restriction, the Metacitrus displayed the unique band between 5.0 and 4.0kb, in addition to four fragments about 5.0, 2.4, 2.15, and 2.0kb, respectively, different from Archicitrus. Also the pattern of total mtDNAs fragment by the treatment of Pst I showed that the distinguishable fragment pat tern was not appeared in Archicitrus(C. iyo Tanaka), but about 6.0, 5.5, 5.0 and 2.Bkb fragments were appeared only in Metacitrus(C. junos Sieb). Therefore it was indicated that two species in intra-subgenus were identical each other, whereas considerable difference was revealed for inter-subgenus.

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Cloning and Characterization of Two Distinct CD3 Genes from Olive Flounder Paralichthys olivaceus

  • Kim, Mu-Chan;Park, Chan-Il
    • Fisheries and Aquatic Sciences
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    • 제8권2호
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    • pp.56-64
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    • 2005
  • Two distinct CD3 homologue genes, $CD3\gamma/\delta\;and\;CD\varepsilon$, were isolated from a olive flounder leukocyte cDNA library and a BAC library. $CD3\gamma/\delta$ consisted of 961 bp encoding 178 amino acid residues, and $CD3\varepsilon$ consisted of 1006 bp encoding 164 amino acid residues. When compared with other known CD3 peptide sequences, the most conserved region of the two olive flounder CD3 chain peptides are the cytoplasmic domain and the least conserved are the extracellular domain. A phylogenetic analysis based on the deduced amino acid sequence grouped the two olive flounder CD3 sequences with $CD3\varepsilon$ and $CD3\gamma/\delta$, respectively. The olive flounder CD3 cluster (consisting of $CD3\varepsilon\;and\;CD3\gamma/\delta$) spans only 10.4 kb. The $CD3\varepsilon\;and\;CD3\gamma/\delta$ genes are oppositely transcribed only 3.8 kb apart. Both olive flounder CD3 genes have five exons. The two olive flounder CD3 genes were predominantly expressed in PBLs, kidney, spleen, and gills.

Detection and Molecular Characterization of a Stolbur Phytoplasma in Lilium Oriental Hybrids

  • Chung, Bong-Nam;Jeong, Myeong-Il
    • The Plant Pathology Journal
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    • 제19권2호
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    • pp.106-110
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    • 2003
  • Stolbur Phytoplasma was detected from Lilium Oriental hybrids showing flattened stem and flower clustering. The presence of phytoplasma was demonstrated using polymerase chain reaction(PCR) assays with phyto-plasma-universal(P1/P6)and stolbur phytoplasma-specific 16F1/R1-S primer pairs amplifying phytoplasma 16S rDNA regions. Nucleotide suquences of the phytoplasma 16S rDNA were determined. Nucleic acid extracted from lily amplified 1.5 kb DNA with a phytoplasma universal primer pair. In nested PCR, 1.1 kb PCR product was obtained using specific primer pair, indicating an isolate of stolbur phytoplasma. Nucleotide sequence of phytoplasma 16S rDNA reported in this study showed 99.5% and 99.1% identities with two known stolbur phytoplamas (16Sr XII-A). Also, it exhibited a sequence homology of 98.0% with phormium yellow leaf (16Sr XII-B), and 97.9% with Australian grapevine yellows (16Sr XII-B). Meanwhile, it showed 98.1% identity with strawberry green petal phytoplama, (16Sr1-C), and 94.7 % with American aster yellows (16Sr1-B). Homology percentage of the 16S rDNA nucleotide sequence suggests that this phytoplama could be classified into the stolbur phytoplasma, subgroup A (16Sr XII-A), as a type strain stolbur.

Cloning and Expression of a Chitinase Gene from Thermoactinomyces vulgaris KFB-C100

  • Yooh, Ho-Geun;Kim, Hee-Yun;Lim, Young-Hee;Cho, Hong-Yon
    • Journal of Microbiology and Biotechnology
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    • 제8권6호
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    • pp.560-567
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    • 1998
  • We have found that Thermoactinomyces vulgaris KFB-Cl00 produces a chitinase. The optimum temperature and pH of the enzyme activity were $55^{\circ}C$ and 6.5. The enzyme was stable after heat treatment at $80^{\circ}C$ for 30 min and stable in acidic and basic conditions (PH 6.0~11.0). The thermostable endo-chitinase from Thermoactinomyces vulgaris KFB-C100 was cloned into the plasmid pBR322 by using E. coli DH5$\alpha$ as a host strain. The positive clone carrying a recombinant plasmid (PKCHI23) with a 4.1-kb fragment containing the chitinase gene was found. The recombinant plasmid was analyzed to determine the essential region for chitinase activity and obtained a 2.3-kb fragment, which was sub cloned into pTrc99A using the PstI and SalI sites to construct pTrc99A/pKCHI23-3. The resulting plasmid exerted high chitinase activity upon transformation of E. coli XL1-Blue cells. Chitinase was overproduced 14 times more in the clone cells than in the wild-type cells and the enzyme was purified to homogeneity. The purified enzyme showed the similar properties as the native chitinase from T. vulgaris in terms of molecular weight and substrate specificity. The catalytic action of the cloned enzyme was an endo type, producing chitobiose as a major reaction product.

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Lactobacillus acidophilus GP4A 박테리오신의 정제, Bacteriolytic 작용 및 생산 관련 Plasmid의 선별 (Purification, Bacteriolytic Action and Plasmid Isolation of Acidocin 4A Produced by Lactobacillus acidophilus GP4A)

  • 한경식;전우민;김영훈;김세헌
    • Journal of Dairy Science and Biotechnology
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    • 제21권2호
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    • pp.114-119
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    • 2003
  • Lactobacillus acidophilus CP4A 균주가 생산하는 acidocin 4A를 정제하고자 ammonium sulphate 침전법 , Octyl-sepharose CL-4B column chromatography, $C_{18}$ Sep-Pak cartridge, $C_{18}$ RP HPLC, HPLC gel filtration을 실시하였고 tricine SDS-PAGE를 통해 약 4.1 kDa의 박테리오신임을 확인하였다. Acidocin 4A의 항균작용 기작을 L. delbrueckii subsp. lactis ATCC 4797을 대상으로 TEM을 이용해 관찰한 결과 세포벽이 해리되고 세포벽사이로 세포내용물이 용출되어 궁극적으로 cell lysis가 일어나는 bacteriolytic 현상을 확인하였다. 또한, acidocin 4A의 생산에 관련된 유전자의 존재 위치를 파악하고자 EtBr을 이용한 curing방법을 실시하였으며 그 결과 약 20 kb 크기의 plasmid에 acidocin 4A 생산과 자체 면역에 관련된 유전자가 존재함을 알 수 있었다.

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홍성 신리 지역 대리암 내 함석류석 변성염기성암의 암석지화학 연구 및 그 지구조적 의미 (Petrochemistry of Garnet-bearing Metabasite in Marble at Shinri area in Hongseong and its Tectonic Implication)

  • 김성원;고희재
    • 암석학회지
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    • 제19권3호
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    • pp.209-225
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    • 2010
  • 남서부 경기육괴 홍성의 동부 예당호 부근의 신리지역은 원생대 후기의 덕정리 화강섬록암-토날라이트, 압쇄 각섬석 정편마암 및 함석류석 변성염기성암을 포함하는 대리암이 분포한다. 이 논문에서는 신리지역의 대리암 내 원생대 후기 함석류석 변성염기성암의 광물화학 및 지화학 자료를 보고하고자 한다. 신리지역의 함석류석 변성염기성암을 $SiO_2$$Na_2O+K_2O$의 분별도에 도시하면 $SiO_2$의 함량은 46.98-51.17 wt% 그리고 $Na_2O+K_2O$의 함량은 1.95-2.85 wt%로 솔레아이트질 서브-알칼라인 현무암의 영역에 점시된다. Zr/Y 대 Zr의 분별도에서도 함석류석 변성염기성암은 서브-알칼라인 현무암의 영역에 점시된다. 신리 지역의 변성염기성암의 희토류 원소 농도를 콘드라이트 조성치에 표준화한 희토류 양상은 중앙해령과 유사한 현무암의 희토류원소 양상을 보여준다. 평균 초생맨틀값으로 미량원소 성분을 표준화하여 도시한 거미 성분도상에서는 신리변성염기성암는 경희토류가 부화되어 있고, 전반적으로 주원소 및 미량원소의 지화학적 특징은 판 내부 환경의 현무암과 유사한 경향을 보여준다. 이들 양상은 기 보고된 비봉과 백동지역의 변성초염기성암과 관련된 고압형 변성염기성암의 중앙해령 현무암과 유사한 호상열도 현무암 혹은 배호분지 현무암의 양상과 상이하다. 신리지역 대리암 내에 포함된 변성염기성암의 광물군과 광물화학을 근거로 계산된 변성 압력-온도 조건은 석류석 중심부에서 9.6-12.7 kb, $695-840^{\circ}C$, 석류석 가장자리에서 9.6-13.6 kb, $630-755^{\circ}C$로 온도가 감소하며 압력이 거의 일정한 변성진화과정을 보여준다. 이들 변성 압력-온도 진화경로는 비봉 및 백동 변성염기성암의 등온하강 후퇴변성작용 경로와는 다름을 알 수 있다. 이들 암체에서 보고된 트라이아스기 변형, 변성 시기와 더불어 원생대 후기의 변성작용과 생성 시기 등을 포함한 보다 상세한 연구가 수행되어져야 할 것이다.

ZNF552, a novel human KRAB/C2H2 zinc finger protein, inhibits AP-1- and SRE-mediated transcriptional activity

  • Deng, Yun;Liu, Bisheng;Fan, Xiongwei;Wang, Yuequn;Tang, Ming;Mo, Xiaoyang;Li, Yongqing;Ying, Zaochu;Wan, Yongqi;Luo, Na;Zhou, Junmei;Wu, Xiushan;Yuan, Wuzhou
    • BMB Reports
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    • 제43권3호
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    • pp.193-198
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    • 2010
  • In this study, we report the identification and characterization of a novel C2H2 zinc finger protein, ZNF552, from a human embryonic heart cDNA library. ZNF552 is composed of three exons and two introns and maps to chromosome 19q13.43. The cDNA of ZNF552 is 2.3 kb, encoding 407 amino acids with an amino-terminal KRAB domain and seven carboxyl-terminal C2H2 zinc finger motifs in the nucleus and cytoplasm. Northern blotting analysis indicated that a 2.3 kb transcript specific for ZNF552 was expressed in liver, lung, spleen, testis and kidney, especially with a higher level in the lung and testis in human adult tissues. Reporter gene assays showed that ZNF552 was a transcriptional repressor, and overexpression of ZNF552 in the COS-7 cells inhibited the transcriptional activities of AP-1 and SRE, which could be relieved through RNAi analysis. Deletion studies showed that the KRAB domain of ZNF552 may be involved in this inhibition.

Isolation and Characterization of the Ribosomal Protein 46 Gene in Drosophila melanogaster

    • Animal cells and systems
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    • 제2권1호
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    • pp.113-116
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    • 1998
  • A cDNA clone coding for ribosomal protein 46 (rp46) which is a component of 60S ribosomal large subunit has been identified from Drosophila melanogaster. A cDNA clone encoding S. cerevisiae rp46 was used as a probe to screen a Drosophila larvae cDNA library. The DNA sequence analysis revealed that the cDNA coding for Drosophils rp46 contains a complete reading frame of 153 nucleotides coding for 51 amino acids. The deduced amino acid sequence showed 71-75% homology with those of other eukaryotic organisms. Northern blot analysis showed that about 1-kb rp46 transcripts are abundant throughout fly development. Whole mount embryonic mRNA in situ hybridization also showed no preferential distribution of the transcripts to any specific region. The chromosomal in situ hybridization revealed that the identified gene is localized at position 60C on the right arm of the second polytene chromosome with a possibility of single copy.

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Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2015년도 춘계학술대회 및 임시총회
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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