• Title/Summary/Keyword: HindIII

Search Result 207, Processing Time 0.02 seconds

Identification of 4-Chlorobenzoate Dechlorinating Bacteria by Simple Plate Assay (평판배지법에 의한 4-chlorobenzoate 탈염소화 세균의 검색)

  • Chae, Jong-Chan;Kim, Chi-Kyung;Min, Kyung-Hee;Park, Yong-Keun
    • Microbiology and Biotechnology Letters
    • /
    • v.23 no.1
    • /
    • pp.104-109
    • /
    • 1995
  • The gene responsible for dechlorination of 4-chlorobenzoate (4CBA) was cloned in E. coli XL1-Blue from Pseudomonas sp. DJ-12. The cloned cell of E. coli Cjl had the hybrid pBluescript SK(+) plasmid, into which about 9.5 kb genomic DNA fragment of PseudOmonas sp. DJ-12 was inserted. The subclone of pCJlOl was constructed by inserting the 3.4 kb EcoRI-HindIII fragment of pCJl into the vector. Those cloned cells could be simply selected by halo formation around the colonies which was the precipitate of AgCl produced by reaction of AgNO$_{3}$ and chloride ion liberated by bacterial dechlorination of 4CBA- Such a plate assay method was standardized by the procedure that the colonies grown for 2 days on the Cl$^{-}$-free plate medium containing 1 mM 4CBA were flooded with 0.1 M AgNO$_{3}$ solution.

  • PDF

Molecular Cloning of a CMCase Gene from Alkalophilic sp. and Its Expression in Escherichia coli

  • Yu, Ju-Hyun;Kong, In-Soo;Kim, Jin-Man;Park, Yoon-Suk
    • Proceedings of the Korean Society for Applied Microbiology Conference
    • /
    • 1986.12a
    • /
    • pp.529.1-529
    • /
    • 1986
  • For isolation of the CMCase gene of the alkalophilic Bacillus sp. strain N-4 to analyze their genetic information for the multicomponents of the cellulase, Bscherichia coli K12 and plasmid DNA pBR322 was used as host-vector system. After the digestion of purified chromosomal DNA and plasmid DNA pBR322 with HindIII, these were ligated. The ligated DND were transformed into Escherichia coli, and recombinant plasmid 107 carried the gene coding for CMCase was constructed. The CMCase produced by Escherichia coli cells containing plasmid DNA pYBC107 was found in the cells as intracellular enzyme and nearly 60% of the total CMCase activity was localized in cellular fraction. Also, the optimum pH for the reaction of CMCase produced by Escherichia coli was appeared at pH .8.0 and the enzyme was stable between pH 7.0 and pH 8.0.

  • PDF

Molecular Cloning and Sequencing of Cell Wall Hydrolase Gene of an Alkalophilic Bacillus subtilis BL-29

  • Kim, Tae-Ho;Hong, Soon-Duck
    • Journal of Microbiology and Biotechnology
    • /
    • v.7 no.4
    • /
    • pp.223-228
    • /
    • 1997
  • A DNA fragment containing the gene for cell wall hydrolase of alkalophilic Bacillus subtilis BL-29 was cloned into E. coli JM109 using pUC18 as a vector. A recombinant plasmid, designated pCWL45B, was contained in the fragment originating from the alkalophilic B. subtilis BL-29 chromosomal DNA by Southern hybridization analysis. The nucleotide sequence of a 1.6-kb HindIII fragment containing a cell wall hydrolase-encoding gene was determined. The nucleotide sequence revealed an open reading frame (ORF) of 900 bp with a concensus ribosome-binding site located 6 nucleotide upstream from the ATG start codon. The primary amino acid sequence deduced from the nucleotide sequence revealed a putative protein of 299 amino acid residues with an M.W. of 33, 206. Based on comparison of the amino acid sequence of the ORF with amino acid sequences in the GenBank data, it showed significant homology to the sequence of cell wall amidase of the PBSX bacteriophage of B. subtilis.

  • PDF

Analysis of Tetracycline Resistance Plasmids and the Conjugative Transfer of Tetracycline Resistancy in Staphylococcus epidermidis (Staphylococcus epidermidis의 Tetracycline 내성 Plasmid의 분석 및 Conjugation에 의한 내성전달)

  • Chung, Jae-Kyu;Park, Mi-Kyung;Kim, Sung-Kwang
    • The Journal of the Korean Society for Microbiology
    • /
    • v.22 no.1
    • /
    • pp.95-99
    • /
    • 1987
  • When tetracycline resistancy were cured by ethidium bromide treatment, some of the cured strains lost the tetracycline resistance plasmid while other strains kept the plasmids. Both strains of lost and remained plasmids were digested with restriction endonuclease Hind III and these cleaved plasmids were compared with that of parent strains, two plasmid remained strains showed same cleavage patterns between parent and cured strains, however, one plasmid lost strain showed dissimilarity with parent strain, but in the other one strain, among 4 plasmid lost colonies, 2 showed same but other 2 showed different patterns compared to parent strain. Tetracycline was transfered by conjugation in on set(Staphylococcus aureus donor versus Staphylococcus epidermidis, recipient) with relative high frequency but the other 2 sets showed a low degree of frequency and the other 2 sets exhibited no transfer.

  • PDF

Pseudomonas sp. W7로부터 Agarase 유전자의 Cloning 및 Escherichia coli에서의 발현

  • Ha, Jeong-Chul;Kim, Gu-Taek;Kim, Sung-Koo;Yu, Ju-Hyun;Kong, In-Soo
    • Microbiology and Biotechnology Letters
    • /
    • v.23 no.6
    • /
    • pp.665-670
    • /
    • 1995
  • A marine bacterium which produces extracelluar agarase was isolated from sea water. Isolated strain was identified as Pseudomonas sp. by the morphological and biochemical properties (1). HindIII restriction fragment of 3.2 kb from Pseudomonas genomic DNA was cloned into pUC19 to obtain recombinant plasmid pJA1 which enables E. coli JM83 to produce agarase. Most of agarase produced in E. coli was secreted into the culture medium. The enzyme (pJA1) showed the highest agarase activity during the stationary phase (20 hrs) of E. coli. The optimum temperature and pH were 40$\circ$C and 7.8, respectively. Restriction gene map anlaysis revealed that it has different restriction pattern with three kind of agarase gene reported.

  • PDF

Cloning and Expression of a Xylanase Gene from Alkali-tolerant Bacillus sp. YA-14 in Escherichia coli (알카리 내성 Bacillus sp. YA-14의 xylanase 유전자 cloning)

  • 유주현;박덕철;정용준;공인수
    • Microbiology and Biotechnology Letters
    • /
    • v.17 no.2
    • /
    • pp.154-159
    • /
    • 1989
  • Chromosomal DNA fragments of Bacillus sp. YA-14 isolated from soil as a potent xylan hydrolyzing bacterium, were ligated to a vector plasmid, pBR322, and used to transfer Escherichia coli HB101 cells. The recombinant plasmid pYDC21 was found to enable the transformants to produce xylanase. pYDC21 was found to contain the 3 kb HindIII fragment originated from the Bacillus sp. YA-14 chromosomal DNA by southern hybridization. The optimum temperature and pH for the reaction of xylanse produced by E. coli (pYDC21) were appeared at 50$_7$C and pH 7.0, respectiveiy. the xylanase enzyme was stable between pH 5.0 and 7.0 and maintained stably up to 4$0^{\circ}C$.

  • PDF

Characteristics of MCPA plasmid isolated from pseudomonas sp. (Pseudomonas에서 분리한 MCPA 플라스미드의 특성)

  • 이영록;최대성;은성호;박영두
    • Korean Journal of Microbiology
    • /
    • v.24 no.4
    • /
    • pp.394-399
    • /
    • 1986
  • From the lysates of the 7 selected strains of Pseulomonas utilizing 2-methyl-4-chlorophenoxyactate as a sole source of carbon and energy, several MCPA plasmids, which encodes genes for the degradation of 2-methyl-4-chlorophenoxyacetate, were isolated, and measured their molecular weight as well as genetic characters such as resistance to antibiotics and degradative ability of other chlorinated herbicides. Transmissibility of the MCPA plasmids, pKU1, pKU15, and pKU17 was tested by conjugation or transformation and the restriction pattern of pKU15 for Pvu II, Hind III, EcoR I, Xho I, Bgl II, and Ava II was analyzed.

  • PDF

MLS Inducible Resistance Mechanism in Bacillus licheniformis EMR-1 -Cloning of erm K, a MLS Resistance Determinant- (Bacillus licheniformis EMR-1에서의 MLS 유도내성 기전 -erm K의 크로닝-)

  • Choi, Eung-Chil;Kwak, Jin-Hwan;Weisblum, Bernard
    • YAKHAK HOEJI
    • /
    • v.32 no.4
    • /
    • pp.213-221
    • /
    • 1988
  • Inducible MLS resistance gene of Bacillus licheniformis specified by erm K was subcloned in Bacillus subtilis and the DNA sequence corresponding to its control region was determined. The determinant erm K was in Pvu II=Hind III fragment, which was 1.3 kb. The leader region is capable of forming a complex series of inverted complementary repeat sequences (ICRS) centering on at least six axes of symmetry, some of them mutually exclusive, in a way that resulted ultimately in post-transcriptional unmasking of the ribosome loading site for methylase synthesis.

  • PDF

No Association of the Human Y Chromosome with Blood Pressure in Korean Male Population

  • Kang, Byung-Yong;Kim, Seon-Jeong;Lee, Kang-Oh
    • Toxicological Research
    • /
    • v.19 no.1
    • /
    • pp.29-31
    • /
    • 2003
  • It has been reported that the genetic variations in the Y chromosome has influence the blood pressure in some Caucasian male populations, but the effect in non-Caucasian population is unclear. In the present study, we examined the relationship between blood pressure and a HindIII RFLP of Y chromosome in 152 unrelated male individuals of ethnically homogeneous Korean origin. There were no significant differences in systolic and diastolic blood pressures between genotype groups, respectively. However, the frequency of A genotype in Korean population was much higher than those of Caucasian populations (P<0.05). Therefore, the results of this study will con-tribute the better understanding the genetic characteristics of Y chromosome in Korean population.

Cloning of the Bacteriocin Gene from Xanthomonas campestris pv. lycines 8ra (콩 불마름병균 Xanthomonas campestris pv. glycines 8ra의 박테리오신 유전자 Cloning)

  • 안응진;조용섭
    • Korean Journal Plant Pathology
    • /
    • v.12 no.2
    • /
    • pp.169-175
    • /
    • 1996
  • 콩 불마름병균 Xanthomonas campestris pv. glycines 8ra는 X. c. pv. vesicatoria에 길항력이 있는 bacteriocin인 glycinecin을 생성 분비한다. Bacteriocin 생성 분비 능력이 있는 콩 불마름병균을 효과적인 생물학적 방제원으로 활용하기 위해서는 좀더 체계적인 연구가 필요하여, bacteriocin 생성에 관계되는 유전자의 분리를 시도하였다. 약 2,000개의 Xanthomonas campestris pv. glycines 8ra cosmid library에서 bacteriocin의 생성 분비 능력을 조사하여 다섯 개의 clone을, pG011, pG0113, pG33과 pG35, 선발하였다. 그중 한 clone pG08을 임의로 선택하여 plasmid DNA를 분리하였다. Plasmid pG08에서 약 6.0 kb의 DNA를 떼어내어 다른 plasmid vector에 넣은 subclone pBL5는 bacteriocin의 생성 분비 능력이 있었다. Plasmid pG08을 제한효소 처리후 다시 접함시켜 만든 몇 개의 subclone과 pBL5의 제한효소 지도를 비교 분석한 결과 약 3.0 kb의 BamHI-HindIII 부분의 DNA가 bacteriocin의 생성에 관계함을 알았다.

  • PDF