• Korean Journal of Microbiology
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• v.24 no.1
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• pp.12-17
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• 1986

A new computer algorithm is described to order the restriction fragments of plasmid DNA which has been cleaved with several restriction endonucleases in single or double digestions rapidly with realistic error rates. The permutation and high weight on small fragments methods construct all logical circular map solutions. The program is written in Apple BASIC and run on an Apple II plus microcomputer with 64K memory. Several examples are presented which indicate the high efficiency of the profram in construction possible restriction map for YEp24.

• Korean Journal of Veterinary Research
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• v.39 no.4
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• pp.797-805
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• 1999

T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.562-567
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• 1999

Single-stranded DNA binding protein (SSB) is well-characterized as having a helix-destabilizing activity. The helix-destabilizing capability of SSB has been re-examined in this study. The results of restriction endonuclease protection assays and titration experiments suggest that the stimulatory effect of SSB on strand exchange acts by melting out the secondary structure which is inaccessible to RecA protein binding; however, SSB is excluded from regions of secondary structure present in native single-stranded DNA. Complexes of SSB and RecA protein are required for eliminating the secondary structure barriers under optimal conditions for strand exchange.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.231-236
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• 1989

The larger segment of double stranded RNA genome from a new serotype of Infectious Pancreatic Necrosis Virus (IPNV), DRT, has been partially cloned at Sma I site in pUC19 and compared with the restriction maps of VR-299 and Sp. Restrction sites found in DRT was distinct and hence a new serotype. The cDNA clones of DRT were about 800, 850, and 1, 400 bp long each and do not share any common restiction site. It is not clear yet if there exist any overlapping sequences among them. This partial cloning, however, was sufficient for the comparison of restriction maps with the other serotypes.

• Biomedical Science Letters
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• v.18 no.1
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• pp.29-34
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• 2012

Streptomycetes are gram-positive filamentous bacteria that are well-known for producing a vast array of bioactive compounds, including more than 70 % of commercially important antibiotics. For the research about Streptomyces sp., the protoplast and electroporation transformation method have been the general techniques for the construction of transformants. However, these techniques have low efficiency and are time-consuming. Another option is intergenic conjugation, which is used for DNA transfer using methylation-deficient E. coli as a DNA donor to avoid the methylated-DNA-dependent restriction systems of actinomycetes. This conjugation method has been widely improved and applied to many other actinomycetes. In this research, an effective transformation procedure for the construction of expression vector by using gateway system was established to avoid limit of restriction enzyme site for cloning of target gene based on transconjugation by Escherichia coli ET12567/pUZ8002 with a pSET152 integration vector.

• Korean Journal Plant Pathology
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• v.10 no.1
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• pp.1-6
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• 1994

Randomly chosen recombinant clones of Fusarium oxysporum were analysed to select useful probes for relatedness analysis of Fusarium oxysporum. Genomic DNA of F. oxysproum f. sp. cubense, digested with HindIII, was ligated to pUC118 and used to transform Escherichia coli strai DH5$\alpha$. Three clones were identified that hybridized to mutiple restriction fragments of some formae speciales of F. oxysporum. These probes detected repetitive sequences in HindIII or EcoRI digested DNAs. Repeated copy clone pFC46, pFC52 and pFC54 showed evident polymorphisms among ten formae speciales of this fungus. Since clone pFC 52 strongly hybridized to multiple EcoRI-digested restriction fragments of f. sp. cubense, it may be useful as a probe for analysis of other genetic characteristics of this forma specialis. The results suggest that our clones might be very useful as probes for relatedness analysis between or within formae speciales of Fusarium oxysporum.

• BMB Reports
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• v.39 no.4
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• pp.361-370
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• 2006

Genetic variation and molecular phylogeny of 22 taxa representing 14 extant species and 3 unidentified taxa of Boesenbergia in Thailand and four outgroup species (Cornukaempferia aurantiflora, Hedychium biflorum, Kaempferia parviflora, and Scaphochlamys rubescens) were examined by sequencing of 3 chloroplast (cp) DNA regions (matK, psbA-trnH and petA-psbJ). Low interspecific genetic divergence (0.25-1.74%) were observed in these investigated taxa. The 50% majority-rule consensus tree constructed from combined chloroplast DNA sequences allocated Boesenbergia in this study into 3 different groups. Using psbA-1F/psbA-3R primers, an insertion of 491 bp was observed in B. petiolata. Restriction analysis of the amplicon (380-410 bp) from the remaining species with Rsa I further differentiated Boesenbergia to 2 groupings; I (B. basispicata, B. longiflora, B. longipes, B. plicata, B. pulcherrima, B. tenuispicata, B. thorelii, B. xiphostachya, Boesenbergia sp.1 and Boesenbergia sp.3; phylogenetic clade A) that possesses a Rsa I restriction site and II (B. curtisii, B. regalis, B. rotunda and Boesenbergia sp.2; phylogenetic clade B and B. siamensis; phylogenetic clade C) that lacks a restriction site of Rsa I. Single nucleotide polymorphism (SNP) and indels found can be unambiguously applied to authenticate specie-origin of all investigated samples and revealed that Boesenbergia sp.1, Boesenbergia sp.2 and B. pulcherrima (Mahidol University, Kanchanaburi), B. cf. pulcherrima1 (Prachuap Khiri Khan) and B. cf. pulcherrima2 (Thong Pha Phum, Kanchanaburi) are B. plicata, B. rotunda and B. pulcherrima, respectively. In addition, molecular data also suggested that Boesenbergia sp.3 should be further differentiated from B. longiflora and regarded as a newly unidentified Boesenbergia species.

• Applied Biological Chemistry
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• v.36 no.3
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• pp.139-145
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• 1993

The methods of isoelectric focusing of 4 isozymes in polyacrylamide horizontal slab gels and restriction fragment length polymorphisms (RFLPs) were applied to characterize the biochemical phenotypes of 19 cultivars of barley. Among 19 barley cultivars screened, 7 esterase, 3 phosphoglucose isomerase, 4 peroxidase and 2 alcohol dehydrogenase isozyme phenotypes were distinguished by isoelectric focusing. When purified DNA of each cultivar was digested with restriction enzyme EcoRV and analyzed its RFLPs with barley DNA markers pMSU 51 or pMSU 71, two distinct RFLP patterns were shown. Based on the four isozymes and two RELP polymorphisms, 19 cultivars of barley were classified into 13 biochemical phenotypes. Phylogenetical relationships among 13 biochemical phenotypes classified were determined using Nei's F-statistics and the phylogenetic dendrogram was constructed.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.3
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• pp.301-307
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• 2005

Genetic variants of Hanwoo mtDNA in the region of cytochrome oxidase subunit I, II and III complex were detected using restriction enzymes. PCR primers were designed based on the bovine mtDNA sequence, and 6 primer sets (Mt4, Mt5, Mt6, Mt7, Mt8 and Mt9) were used. A total of 20 restriction enzymes were used, and 6 restriction enzymes, which were Hinf I, Pvu II, Rsa I, Eco RI, Bgl II, and Msp I, showed genetic polymorphisms. Significant associations between genetic variants and weight traits were observed at WT15 (p<0.05) and WT18 (p<0.01) with Pvu II for Mt9, Bgl II for Mt6 and Rsa I for Mt8 segments in the region of cytochrome oxidase subunit complex. Significant associations were also observed at Mt9-Pvu II and Mt6-Bgl II segments for WT9 (p=0.01), WT12 (p=0.02), respectively. These results suggest that genetic variants of mtDNA in the region of cytochrome oxidase subunit complex may be candidate segments for improvement of animal growth as weight traits.

• Korean Journal of Plant Taxonomy
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• v.34 no.1
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• pp.43-61
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• 2004

Molecular taxonomic studies were conducted to evaluate interspecific relationships in Korean Viola 34 taxa including two Japanese populations using RAPD(randornly amplified polymorphic DNA), ISSR(inter simple sequence repeat) and PCR-RFLP(restriction fragment length polymorphism) analysis. Only six and four primers out of 40 arbitrary and 12 ISSR primers were screened for 34 taxa, and were revealed 70 (98.6%) and 28 (96.6%) polymorphic bands, respectively. Fifteen restriction endonucleases produced 80 restriction sites and size variations from the large single copy region of cpDNA, 16 (20%) of which were polymorphic. The separate analyses from the RAPD, ISSR and PCR-RFLP data were incongruent in the relationships among 34 taxa, but combined data was in accordance with previous infrageneric classification system based on morphological characters, especially the subsection and series level. Section Chamaemelanium placed between subsect. Patellares and Vagimtae of section Nomimium was not formed as a distinct group. Viola alb ida complex including three very closely related taxa was recognized independent group within subsect. Patellares in combined data tree. This result strongly suggested that they should be treated to series Pinmtae. RAPD analysis was very useful to clarify the interspecific relationships among the species of Korean Viola than ISSH and PCR-RFLP analyses.