• Reproductive and Developmental Biology
• /
• v.33 no.1
• /
• pp.19-24
• /
• 2009

This study was conducted to investigate the specific expression genes in the cloned bovine tissues. Donor cells, cloned tissues were analysed by RAPD-RFLP method. The results were detected three genes (CH-U7B, CH-U7M and CH-U7P) in the cloned fetus. It was found a single copy genes by southern hybridization. Sequence analysis of CH-U7M gene was shown 99% homology to a previously reported EST from a cloned bovine fetus. The putative ORF was encode a protein of hydrophobicity index 0.03. Semi-quantitative RT-PCR by using the CH-LS001 specific primer was remarkably detected in the lung tissue of cloned fetus. Further investigation of these genes may provide one of the key information to explain the early death, abnormal fetus, large off-spring and the low pregnancy rate in the production of cloned bovine.

• Journal of Veterinary Clinics
• /
• v.21 no.2
• /
• pp.102-108
• /
• 2004

Orf virus (ORFV), a member of genus Parapoxvirus (family-Poxviridae), a causative agent of contagious ecthyma in sheep and goat leading to a condition commonly known as vesicular dermatitis. Recently, twelve goats from Iksan in Jeonbuk province were observed with clinical signs like necrotic vesicular lesions around the mucosa of mouth, nasal cavity, eye, ear, teats, abdomen and groin. Based on these clinical symptoms, contagious ecthyma infection was suspected. The skin scrapping was collected from lesions for isolation of DNA and subsequent PCR amplification of ORFV specific 235 bp region of B2L gene. All of the samples were found positive by PCR analysis. Sequencing and further phylogenetic analysis of the PCR product revealed 100% identity to Japan isolate of ORFV (Okinawa, GenBank accession number AB080769), and showed 99.6% of similarity to New Zealand strain (NZ-2, GenBank accession number U06671). It was concluded that ORFV strain detected in the present study is homologous to Japan isolate and New Zealand strain. The PCR test based on amplification of B2L gene is a highly useful tools for rapid and specific diagnosis of contagious ecthyma.

• Korean Journal of Plant Resources
• /
• v.17 no.3
• /
• pp.289-296
• /
• 2004

A cDNA clone (GenBank accession no. CF924621) homologous to aluminum induced protein gene was isolated and characterized from Codonopsis lanceolata (ClAIP). The ClAIP is 906 nucleotides long and has an open reading frame of 711 bp with a deduced amino acid sequence of 236 residues. The ClAIP shows high homology to A. marina (84%), G. hirsutum(83%), V. radiata (83%), A. thaliana (80%), B. nap us (78%) and T. aestivum (68%). The deduced amino acid sequence of ClAIP also has homology to the N-terminal end of plant Asn synthetase. This region does not contain the active sites of the enzyme and the significance of this conservation is currently not clear. To investigate the expression of ClAIP against several heavy metal stresses, we treated the sliced tap root of C. lanceolata with various heavy metals. The expression of ClAIP was increased by 25 uM $Al_2$(SO$_3$)$_4$ in proportion to incubation time and also increased by 50 uM CdCl$_2$.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.4
• /
• pp.670-677
• /
• 2010

An alkaline pectate lyase, Bsp165PelA, was purified to homogeneity from the culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme showed a specific activity as high as 1,000 U/mg and had optimum activity at pH 11.5 and $50^{\circ}C$. It was composed of a single polypeptide chain with a molecular mass of 42 kDa deduced from SDS-PAGE, and its isoelectric point was around pH 6.0. It could efficiently depolymerize polygalacturonate and pectin. Characterization of product formation revealed unsaturated digalacturonate and trigalacturonate as the main products. The pectate lyase gene (pelA) contained an open reading frame (ORF) of 1,089 bp, encoding a 36-amino acids signal peptide and a mature protein of 326 amino acids with a calculated molecular mass of 35.943 Da. The deduced amino acid sequence from the pelA ORF exhibited significant homology to those of known pectate lyases in polysaccharide lyase family 1. Some conserved active-site amino acids were found in the deduced amino acid sequence of Bsp165PelA. $Ca^{2+}$ was not required for activity on pectic substrates.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 2004.06a
• /
• pp.273-277
• /
• 2004

$\alpha$-L-Arabinofuranosidase ($\alpha$-L-AFase, EC 3.2.1.55) was isolated from hyperthermophilic microorganism, Thermotoga maritima. The open reading frame (ORF) of $\alpha$-L-AFase gene is 1,455 bp long and encodes 484 amino acid residues with a molecular weight of 55,265 Da. The ORF of $\alpha$-L-AFase gene was introduced into the E. coli expression vector, $_p/RSET-B, and overexpressed in E. coli BL21. The purified recombinant $\alpha$-L-AFase showed the highest activity at 10$0^{\circ}C$ and pH 5.5. The purified enzyme appeared to have no metal cofactor requirement. The Km and specific activity values of the recombinant enzyme were 0.99 mM and 1,200 U/mg on p-nitrophenyl-$\alpha$-L-arabinofuranoside. It released only L-arabinose from sugar beet arabinan, sugar beet debranched arabinan and oat spelts arabinoxylan but had no activity onarabinogalactan and gum arabic. This result suggests that L-arabinose could be produced from natural polysaccharides using this enzyme. Mutant enzymes which Glu26, Glu172 and Glu281 residues were replaced to alanine, aspartic acid or glutamine caused Kcat to decrease by a factor of between 10$^3$ and 10$^4$. Glu172 and Glu281 residues of $\alpha$-L-AFase are seemed to be the acid/base and nucleophile in catalytic reaction, respectively, and Glu26 is supposed to playa key role in substrate binding.ng.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.9
• /
• pp.1460-1468
• /
• 2007

In this study, an approx. 2.5-kb gene fragment including the catalase gene from Rhodospirillum rubrum S1 was cloned and characterized. The determination of the complete nucleotide sequence revealed that the cloned DNA fragment was organized into three open reading frames, designated as ORF1, catalase, and ORF3 in that order. The catalase gene consisted of 1,455 nucleotides and 484 amino acids, including the initiation and stop codons, and was located 326 bp upstream in the opposite direction of ORF1. The catalase was overproduced in Escherichia coli UM255, a catalase-deficient mutant, and then purified for the biochemical characterization of the enzyme. The purified catalase had an estimated molecular mass of 189 kDa, consisting of four identical subunits of 61 kDa. The enzyme exhibited activity over a broad pH range from pH 5.0 to pH 11.0 and temperature range from $20^{\circ}C$ to $60^{\circ}C$C. The catalase activity was inhibited by 3-amino-1,2,4-triazole, cyanide, azide, and hydroxylamine. The enzyme's $K_m$ value and $V_{max}$ of the catalase for $H_2O_2$ were 21.8 mM and 39,960 U/mg, respectively. Spectrophotometric analysis revealed that the ratio of $A_{406}$ to $A_{280}$ for the catalase was 0.97, indicating the presence of a ferric component. The absorption spectrum of catalase-4 exhibited a Soret band at 406 nm, which is typical of a heme-containing catalase. Treatment of the enzyme with dithionite did not alter the spectral shape and revealed no peroxidase activity. The combined results of the gene sequence and biochemical characterization proved that the catalase cloned from strain S1 in this study was a typical monofunctional catalase, which differed from the other types of catalases found in strain S1.

• Proceedings of the Korean Institute of Intelligent Systems Conference
• /
• 2004.04a
• /
• pp.255-258
• /
• 2004

본 연구는 유전자 기능예측에 있어서 유사성 검색과 비교유전체학이 가진 한계를 극복하기 위하여 9종의 Human Herpesvirus를 대상으로 COG와 계통유전학적 방법을 적용하여 향상된 유전자 기능예측을 하고자 하였다. COG의 방법을 이용하여 114 HCOGs (Human Herpesvvirus COGs)를 구축하고, HCOGs를 바탕으로 유전자 컨텐츠트리를 제작하였다. 이 트리를 통하여 각 HCOG는 $\alpha$-특이적 그룹, $\beta$-특이적 그룹, $\alpha$, $\beta$, ${\gamma}$ -특이적 그룹 중 하나에 속함을 보였다. 계통유전체학의 적용을 위하여 u, $\beta$, ${\gamma}$ -특이 그룹에 속하는 ORF중 DNA polymerase를 이용하여 종트리를 제작하였다. SDI (Speciation and Duplication) 알고리즘을 통하여 148개의 당단백질에서 47개의 복제점을 예측하였고, 초기 HCOG의 제작에서 제외되었던 7 ORF는 당단백질과 관련된 5개의 HCOG로 재 정의 하였다. 이 연구를 통하여 COG는 ortholog 그룹을 를러스터링하는데 효과적인 방법이며, 이를 더욱 보완할 수 있는 방법으로 비교유전체학이 사용될 수 있음을 확인하였다. 이는 비교유전체학의 방법과 계통유전체학적 방법을 조화시켜 유전자 기능 예측을 보완할 수 있음을 보여 주었다.

• Asian-Australasian Journal of Animal Sciences
• /
• v.26 no.8
• /
• pp.1057-1064
• /
• 2013

p70 ribosomal S6 kinase (p70S6K) can integrate nutrient and growth factor signals to promote cell growth and survival. We report our molecular characterization of the complementary DNA (cDNA) that encodes the goat p70S6K gene 40S ribosomal S6 kinase 1 (S6K1) (GenBank accession GU144017) and its 3' noncoding sequence in Inner Mongolia Cashmere goats (Capra hircus). Goat S6K1 cDNA was 2,272 bp and include an open reading frame (ORF) of 1,578 bp, corresponding to a polypeptide of 525 amino acids, and a 694-residue 3' noncoding sequence with a polyadenylation signal at nucleotides 2,218 to 2,223. The relative abundance of S6K1 mRNA was measured by real-time PCR in 6 tissues, and p70S6K expression was examined by immunohistochemistry in heart and testis. The phosphorylation of p70S6K is regulated by mitogen-activated protein kinase (MAPK) signaling in fetal fibroblasts.

• 한국생물공학회:학술대회논문집
• /
• 2000.11a
• /
• pp.655-659
• /
• 2000

A 11.5-kb DNA fragment containing an endo-inulinase gene was cloned from Xanthomonas oryzae #5. It contained a single open reading frame of 3,999bp, encoding a polypeptide composed of signal peptide of 32 amino acids and mature protein of 1,301 amino acids. From the comparison of amino acids sequences with fructan hydrolases, inulinase, levanase and CFTase, the sequence of the endo-inulinase had highly homology of 72% with CFTase of B. circulans, and six highly conserved regions including the ${\beta}-fructosidase$ motif were found.

• Nuclear Engineering and Technology
• /
• v.40 no.3
• /
• pp.183-190
• /
• 2008

Pyroprocessing is the optimal means of treating spent metal fuels from metal fast fuel reactors and is proposed as a potential option for GNEP in order to meet the requirements of the next generation fuel cycle. Currently, efforts for research and development are being made not only in the U.S., but also in Asian countries. Electrorefining, cathode processing by distillation, injection casting for fuel fabrication, and waste treatment must be verified by the use of genuine materials, and the engineering scale model of each device must be developed for commercial deployment. Pyroprocessing can be effectively extended to treat oxide fuels by applying an electrochemical reduction, for which various kinds of oxides are examined. A typical morphology change was observed following the electrochemical reduction, while the product composition was estimated through the process flow diagram. The products include much stronger radiation emitter than pure typical LWR Pu or weapon-grade Pu. Nevertheless, institutional measures are unavoidable to ensure proliferation-proof plant operations. The safeguard concept of a pyroprocessing plant was compared with that of a PUREX plant. The pyroprocessing is better adapted for a collocation system positioned with some reactors and a single processing facility rather than for a centralized reprocessing unit with a large scale throughput.