Through an application of plasmid capture system (PCS) to Bacillus thuringiensis plasmid DNAs, we acquired 21 polymorphic clones of putative genomic DNA of bacteriophage. The genome size of phage 1-3 (PhBT1-3) was determined to be 46,517 base pairs (bp) with 35.43% G + C content and 83% coding region. Sixty-five putative open reading frames (ORFs) with more than 50 codons were found in the new phage genome. In accordance with this genome finding, the phage particles and its DNA were confirmed from the supernatant of B. thuringiensis 1-3. Morphological characterization and infectivity assay demonstrated that PhBT1-3 belongs to the family Siphoviridae and it showed infectivity to three B. thuringiensis type strains, galleriae, entomocidus, and morrisoni. Based on these results, we screened the existence of phages in B. thuringiensis type strains by PCR with terminase small subunit-specific primers. Ten of 67 type strains showed PCR products and the similarity of those putative amino acids was more than 70%. Furthermore, we verified the existence of various shaped phages from the supernatants of 10 B. thuringiensis type cultures. In conclusion, we characterized a putative genome of phage, PhBT1-3 from B. thuringiensis 1-3, and confirmed the distribution of phages in the group of 67 B. thuringiensis type strains.

Plasmid capture systems (PCS) facilitate cloning and manipulation of circular double-stranded DNA. We recently developed an improved PCS (PCS-LZ) to clone relatively large DNA molecules of 30-150 kb. The PCS-LZ donor consists of a mini-F replicon and a kanamycin resistance marker between Tn7 left and Tn7 right ends. Both the replicon and marker gene of the PCS-LZ donor are transferred into target plasmid DNAs by in vitro transposition, followed by replication in E. coli. Colonies are tested for lacZ expression by blue/white screening. Circular DNAs were obtained from plasmids of Bacillus thuringiensis, genome segments of Cotesia glomerata bracovirus and polymorphic genomes of Autographa californica nucleopolyhedrovirus. PCS-LZ is a powerful tool for use in genomic analysis and mutagenesis in invertebrate pathology, and we are extending its application to include vertebrate research.

Previously, we found that expression by translational fusion of the polyhedrin (Polh)-green fluorescence protein (GFP) led to the formation of granular structures and these fluorescent granules were easily precipitated by high-speed centrifugation. Here, we developed an easy, fast, and mass purification system using this baculovirus expression system (BES). An enhanced GFP (EGFP) fused with Polh gene at the N-terminus including an adaptor and enterokinase (EK) site between Polh and EGFP was expressed in Sf9 cells. The cells infected by AcPolhEKA-EGFP produced fluorescent granules. The EGFP fusion protein was purified from granule-containing cells according to three steps; cell harvest, sonication and EK digestion. Through the final enterokinase digestion, EGFP was presented mainly in the supernatant (93.3%) and the supernatant also showed a pure EGFP band. These results suggest that the combined procedure of Polh fusion expression and enterokinase digestion can used for the rapid and easy purification of other proteins.

Plasmid capture system (PCS) was developed for easy cloning and manipulation of circular double-stranded DNA from various sources. Recently, we improved PCS system (named PCS-LZ) to clone relatively large-sized DNA molecules (30-150 kb). PCS-LZ donor consists of a Mini-F replicon and a kanamycin resistance marker between Tn7L and Tn7R regions. Both replicon and marker gene of PCS-LZ donor are transferred into target plasmid DNAs by in vitro transposition and the transposed DNAs can replicate in E. coli cells by transformation. White/blue screening by LacZ expression is also available to avoid backgrounds. Up to now, we acquired various circular DNA clones from four sources such as plasmids of B. thuringiensis, bacteriophage genome isolated from B. thuringiensis, genome segments of Cotesia glomerata bracovirus, and polymorphic genomes of Autographa californica nucleopolyhedrovirus. Among them, interestingly, the genome clones of bacteriphage (Ph1-3) were screened from the PCS transposition with plasmids of B. thuringiensis 1-3 strain. The genome of Ph1-3 was fully sequenced (46517 bp) and open reading frames were analyzed. In accordance with this genome finding, the phage particles and its DNA were confirmed from the supernatant of B. thuringiensis 1-3. Ph1-3 showed infectivity to B. thuringiensis type strains such as subsp. galleriae, entomocidus, and morrisoni. Based on these results, we screened the existence of phage in B. thuringiensis type strains by PCR with terminase small subunit-specific primers. Ten of 67 type strains showed PCR products and their sequence similarity was more than 70%. Conclusively, we expect this PCS-LZ system would be a powerful tool for genomic analysis and mutagenesis study at the area of invertebrate pathology and further its application will be enlarged to the vertebrate pathology area.

우라늄 변환시설 가동 중 발생하여 라군(lagoon)에 저장중인 방사성 슬러지 폐기물에 대한 처리는 시설 해체과정에서 매우 중요한 업무 중 하나이다. 슬러지 구성성분 중 다량을 차지하는 질산암모늄의 폭발 위험성 등으로 인해 미생물을 이용한 질산염의 분해는 질산염을 안정적으로 처리할 수 있는 효과적인 방법이라 할 수 있다. 본 연구에서는 라군 슬러지의 약 60 wt%를 차지하는 질산염을 혐기성 균주의 하나인 Pseudomonas halodenidificans를 이용하여 탈질하기위한 공정 변수에 대한 영향을 평가하였다. 온도, 질산염 농도, 전자공여체의 영향, C/N 비율, 초기 접종하는 균주의 비율, pH등의 공정변수에 대하여 실험한 이번 결과는 향후 연속식 공정 설계를 위한 기초 자료로 사용될 것이다.

우라늄 변환시설의 라군 슬러지에 함유된 질산염의 안정적 처리를 위해 물 첨가 용해를 실시한 뒤, 여과 케이크의 안정화 특성에 대하여 알아보았다. 물 용해에 의해 대부분의 질산염은 고농도 질산염 용액으로 제거되었으므로, 여과 케이크의 열분해는 에서 하나의 단계로 수행하였다. Muffle furnace를 이용하여 에서 5시간동안 여과 케이크의 열분해를 실시한 결과 라군 1 슬러지에 포함된 U은 의 열분해와 함께 의 형태로 안정화 되었다. 라군 2 열분해 잔류물의 경우에는 열분해 시 생성된 CaO가 냉각과정에서 수분과 반응하여 로 전환되는 것을 TG/DTA 분석과 XRD 분석을 통해 확인할 수 있었지만, 처분장에서 대기중 노출이나 지하수의 침출 등에는 안정한 화합물로 알려져 있으므로, 특별한 첨가제의 첨가 없이 단순 열분해 후 처분이 가능할 것으로 판단된다.

Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, H+-transporting ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.

Biomass as a renewable energy source has several limitations in terms of the potential for steady supply and its thermal characteristics. This study conducted a thermal weight change analysis and determined its kinetics to address this problem. Sawdust was chosen as the biomass, and PE and PP were the plastics used. Based on the result of thermogravimetric analysis (TGA), the kinetic characteristics were analyzed using Kissinger, Ozawa, and Friedman methods, which are the most common methods used to obtain reaction coefficients and activation energy. The methods used to determine the thermal degradation kinetics were considered feasible for evaluating the pyrolytic behavior of the materials tested. The experimental results of this study provided insights into mixed biomass/plastics pyrolysis kinetics and their optimal operation conditions.

A new Japonica rice variety 'Weonhwangbyeo', released in 1999, was developed from the cross Milyang96//Milyang95/Iri390 made at the Yeongdeog Substation of National Yeongnam Agricultural Experiment Station, RDA. The average heading date of Weonhwangbyeo i

A new wheat variety, 'Anbaekmil' was developed from the cross between Saemil and Geurumil by the wheat and barley breeding team of the National Yeongnam Agricultural Experiment Station (NYAES) in 2001. A promising line, YWM2078- 3B-2-3-7, was selected in