• Title/Summary/Keyword: ${\lambda}$ DNA

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Polyamine Group Assembled Silica Coated Ferrite Nanoparticle for Lambda DNA Detection

  • Park, Moo-Eon;Chang, Jeong-Ho
    • Proceedings of the Korean Powder Metallurgy Institute Conference
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    • 2006.09b
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    • pp.1193-1194
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    • 2006
  • The magnetic ferrite nanoparticles were synthesized and coated by silica precursor in controlling the coating thicknesses and sizeses. The surface modification was performed with amino-functionalized organic silanes on silica coated magnetic nanoparticles. The use of functionalized self-assembled magnetic ferrite nanoparticles for nucleic acid separation process give a lot of advantages rather than the conventional silica based process.

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Isolation of 5'-Untranslational Region of Trout Cyp1A1 Gene

  • Roh, Yong-Nam;Sheen, Yhun-Yhong
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.450-455
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    • 1996
  • The genomic DNA was prepared from trout liver which was treated with 3-methycholanthrene, and cloned into lambda EMBL3 at BamHl site. The genomic library was constructed via infections of these recombinant phages into E. coli K802, and screened by the most $5^I$-portion of trout CYP1A1 cDNA. After the screening of $10^9$ clones of the amplified library, 12 positive clones were isolated, and subjected to further screenings. The results of southern blot hybridization of genomic DNA prepared from the positive clone showed the presence of a single gene of CYP1A1, and 3.5 Kb PstI fragment that hybridizes with the most $5^I$-region DNA of CYP1A1 cDNA. The restriction map of PstI fragment was determined by the restriction digestion with various enzymes. The nucleotide sequence of the upstream genomic DNA of CYPIAI was determined by DNA sequencing of exonuclease III unidirectionally deleted PstI fragment DNA using $[^{35}/S]$dATP. This paper presented the upstream genomic DNA of CYP1A1 contained a part of coding region which was about 351 base pairs (from ATG to PstI site at 3563).

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Chaperon Effects of Campylobacter jejuni groEL Genes Products in Escherichia coli (Campylobacter jejuni의 groEL 유전자 산물의 대장균에서의 Chaperon효과)

  • Lim, Chae-Il;Kim, Chi-Kyung;Lee, Jae-Kil
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.47-52
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    • 1994
  • The cells of Campylobacter jejuni heat-shocked at 48${\circ}C$ for 30 min synthesized the heat shock proteins of HSP90, HSP66 and HSP60. Those heat shock proteins were found to correspond to the heat shock proteins of HSP87, HSP66 (DnaK), and HSP58 (GroEL) of E. coli, respectively. By Southern blot analysis of the chromosomal DNAs of C. jejuni with groESL and dnaK genes of E. coli as DNA probes, the heat shock genes of C. jejuni which are homologous to the E. coli groESL and dnaK genes were found to exist in the chromosomal DNA. The genomic libraries of C. jejuni were constructed with the cosmid vector pWE15 and the groEL gene of C. jejuni were cloned in E. coli B178 groEL44 temperature senstive mutant. The hybrid plasmid (pLC1) was inserted with the DNA fragment (about 5.7kb in size) containing the groEL gene. E. coli groEL44 mutant cell transformed with the pLC1 could grow at 42${\circ}C$ by synthesizing the HSP60 of C. jejuni and regained the susceptibility to the ${\lambda}$ vir phage by expression of the groEL gene in the cloned cells. These indicated that the groEL products of C. jejuni had chaperon effects by synthesizing the heat shock proteins in the cloned cells of E. coli.

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Characterization of a Restriction Endonuclease, SdiI from Streptomyces diastatochromogenes (Streptomyces diastatochromogenes로부터 분리된 SdiI의 특성에 관한 연구)

  • Bae, Moo;Song, Eun-Sook;Hwang, Hye-Yeon;Yim, Jeong-Bin
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.301-305
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    • 1994
  • In catalytic properties of the restriction enonuclease, SdiI, which was purified from Streptomyces diastatochromogenes, this enzyme was active at wide range between pH 7.0 and 12.5, and up to $60^{\circ}C$ and 500 mM of NaCl concentration. It was stable between 20^{\circ}C$ and $60^{\circ}C$, and essentially requires $MgCl_2$ for endonuclease activity. The restriction map of lambda DNA which was obtained by double digestion with various enzymes suggested SdiI to be an isoschizomer of XhoI. From the determination of restriction site based on DNA sequencing method, recognition and cleavage specificity of SdiI was concluded as: 5‘-C${\downarrow}$TCGA G-3' 3'-G AGCT${\uparrow}$C-5'

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A Plausible Method for the Diagnosis of Genetic Disorders Using Full Length cDNA

  • Hur, Hyang-Suk;Lee, Young-Won;Park, Hyoung-Woo;Kim, Myoung-Hee
    • Biomedical Science Letters
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    • v.7 no.1
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    • pp.1-5
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    • 2001
  • A cDNA of coagulation Factor IX gene has been screened from the $\lambda$gt11 human fetal liver cDNA library, and used to construct a 2.8-kb full length cDNA after recombining with the N-terminal fragment from pTZ-FIX. Human genomic DNA was isolated, digested with the restriction endonucleases, TaqI, EcoRI, and HindIII, and Southern hybridization was performed using the full length factor IX cDNA as a probe. The hybridized bands generated by the restriction endonucleases were the followings: TaqI, 0.3, 1.0, 1.6, 1.8, 2.7, 3.7, and 5.3 kb bands; EcoRI, 1.8, 4.8, 4.9, 5.5, 6.8, and 12.6 kb bands; HindIII, 4.1, 4.4, 5.2, 5.8, 7.6, and 12.5 kb bands. When the Southern bands were physically mapped along the genome, about 50-kb continuous region harboring almost all of the genomic region of Factor Ⅸ gene was covered. These results suggest a possibility of using an exonal cDNA probe to diagnose abnormalities including large deletions, insertions, and rearrangements along the genome, if there is any.

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Amino Acid Sequence Homology of Hybrid Poplar O-methyltransferuse Involved in Lignin Biosynthesis

  • Park, Young-Goo;Sul, Ill-Whan;Shin, Dong-Ill;Park, Jang-Won;Park, Hee-Sung
    • Journal of Plant Biotechnology
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    • v.3 no.3
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    • pp.131-134
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    • 2001
  • In $\lambda$-Zap II vector system, a cDNA library was constructed for the developing secondary xylem mRNA from hybrid poplar, Populus nigra x maximowiczii. A cDNA clone of 1.5 kb in size, pOMTB1.4 encoding a lignin-bispecific O-methyltransferase was screened by plaque hybridization using a probe of 540 bp cDNA amplified by polymerase chain reaction from the cDNA library and identified by nucleotide sequencing. Its nucleotide sequence contains one open reading frame of 366 amino acids. The deduced amino acid sequence in comparison with that of Populus tremuloides showed the differences of 9 amino acids and revealed 85-99% homology among alfalfa, poplar and aspen.

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Characterization of BmaI endonuclease from bacillus macerans ATCC 8244 (Bacillus macerans의 BmaI endonuclease의 특성에 대한 연구)

  • 권용태;전희숙;노현모
    • Korean Journal of Microbiology
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    • v.26 no.1
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    • pp.1-5
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    • 1988
  • The esolation and characterization of a new type II restriction endounclease, BamI, from Bacellus macerans ATCC 8244 were described. BmaI endonuclease was partially purified by procedures of ammonium sulfate fractionation, DEAE-cellulose and phosphocellulose chromatographies. This enzume recognized one site on pBR322 DNA, two sites on Bluescribe DNA, three sites on $\lambda$DNA and no site on SV 40 DNA. The same cleavage patterns for vareius DNAs as PvuI indicated that BamI is an isoschisomer of PvuI whose recognition sequence is 5'-CGATCG-3'. The optimal pH for the BmaI endonuclease activity was about 7.0 and optimal NaCl concentration was about 100mM. Manganese ion could partially replace magnesium as a cofactor, but calcium could not at all.

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Cloning and Transcriptional Fusion with lacZ of a Gene (exo) Required for Exo-polysaccharide Synthesis in Rhizobium fredii USDA191 (Rhizobium fredii USDA191의 체외다당류 합성관련 유전자(exo)의 클로닝 및 lacZ와의 융합)

  • 정완석;고영환
    • Korean Journal of Microbiology
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    • v.31 no.1
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    • pp.27-36
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    • 1993
  • Rhizobium fredii USDA191 은 대기 중의 질소를 환원하여 식물체의 생육에 필요한 질소원을 공급해주는 세균으로 다량의 체외 다당류를 합성한다. 전위요소 Tn5의 삽입에 의한 돌연변이 유도로 다당류결핍 변이주 R. fredii YKL293 가 분리되었으며 이 변이주로부터 Tn5 에 인접한 DNA 단편이 pUC19 에 클로닝되었고(plyk5293),이 DNA 단편을 탐침으로 하여 .lambda.NM1149 에 구성되 USDA191 genomic library 로부터 야생형체외다당류 합성관련 유전자(exo) 를 함유한 클론 .lambda. NM1149 22E 를 plaque 혼성화에 의하여 분리하였다. 클론 NM1149.22E 에 들어있는 exo 유전자를 pBR322 에 옮겨서 pJW33을 만들고, 재조합체 pJW33 을 Escherichia coli POII734 에 도입시켜 lacZ 구조유전자를 함유한 MudI 1734 가 exo 유전자의 프러모토와 융합되어 lacZ 구조유전자의 전사가 이루어지도록 하였다. 위와 같이 만들어진 재조합체 플라스미드 pUM21을 함유한 E. coli JM83 은 .betha.-galactosidase 를 합성하였으며, 야생형 tacZ 유전자를 갖고 있는 E. coli LE392 에 비해서 14-25배 정도 낮은 역가를 보였다.

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Therrnosensitive $cI_{857}$ Repressor Overproduction by tac Promoter in General E. coli (일반 E.coli에서 tac Promoter에 의한 온도감수성 $cI_{857}$ Repressor의 대량생산)

  • 강상모;권태종;정호권
    • Microbiology and Biotechnology Letters
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    • v.19 no.1
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    • pp.45-51
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    • 1991
  • Inserting the $cI_{857}$ structural gene in the downstream of the tac promoter for the overproduction of $cI_{857}$ repressor protein was studied. DNA fragment containing $cI_{857}$ ; repressor gene was amplified by using plasmid pUC12, and partially digested with HphI. Only the $cI_{857}$ structural gene isolated was inserted in the downstream of the tac promoter. Plasmid pDR540- $cI_{857}$ having the tuc promoter-$cI_{857}$ structural gene insert could be isolated by the immunity of cells resistant at $30^{\circ}C$ and cell lysis at $42^{\circ}C$ to $\lambda$ phage $cI_{90}$. The amount of $cI_{857}$ repressor as 17% of total cellular protein were produced by using general E. coli as well as $lacI^q$ JM103 having this plasmid.

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