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조애리,문희경 德成女子大學校 藥學硏究所 2003 藥學論文誌 Vol.14 No.1
Acemetacin is a new potent non-steroidal anti-inflammatory compound which is used for the treatment of arthritis and rheumatic diseases. To establish an assay condition for a Bioequivalence study of acemetacin, a column liquid chromatographic (HPLC) method using UV detection for the determination of acemetacin and its metabolites indometacin in blood has been developed. The HPLC apparatus used in this study was a Waters 2695 Separation Module (Alliance) system. Samples were analyzed with Dual-λ-Absorbance Detector (UV254 nm). C_(18) (250×4.6mm, Luna 5μm, Phenomenex) column which maintained at 40℃, provided a good resolution. The mobile phase was composed of 0.02M 인산용액 (pH 4.5):MeOH=45:55. The flow rate was 1.4ml/min. Typical retention times with Luna column were 23, 31 and 34 min for flubiprofen (internal standard), acemetacin and indometacin, respectively. For the data manipulation, Waters Millennium program was employed. Calibration curves for the determination of acemetacin and indometacin in plasma showed a good linearity at a concentration range from 100ng/ml to 4000ng/ml(r²=0.999). The lower detection limits for both compounds were 100ng/ml.
Rat Brain cDNA Library로부터 SNAP-25 유전자의 클로닝
조애리,지영미,유민,이순철,유관희 대한의생명과학회 2000 Journal of biomedical laboratory sciences Vol.6 No.1
SNAP-25는 presynaptic plasma membrane에 위치하는 단백질로서 synaptic vesicle의 docking과 fusion에 있어서 매우 중요한 역할을 한다. 생쥐 SNAP-25 유전자와 99%의 높은 homology를 갖고 있는 Z2 cDNA를 probe로 사용하여 쥐의 뇌 cDNA library에서 SNAP-25 유전자를 screening 하였다. 그 결과 6개의 양성 클론을 분리해 냈으며, 이들 각각을 S1, S2, S3, S4, S5, S6으로 명명하였다. 이중에서 생쥐 SNAP-25와 가장 높은 homology를 보여 주고 있는 S5 클론을 선택하여 염기서열을 분석하였다. 2,100 bp의 염기서열로 구성된 쥐 SNAP-25 cDNA는 206개의 아미노산을 coding하는 618 bp의 open reading frame을 가지고 있으며, ORF는 209∼211 bp에 위치하는 AUG codon에서 시작하여 827∼829 bp에 위치 하는 stop codon TAA에서 끝난다. 3' untranslated region에서는 28과 19개의 CA 반복 염기서열을 보여주고 있었으며, SNAP-25 peptide sequence에서 4개의 cystein residues는 84∼91에 위치하고 있었으며, amino terminus부분에서 amphipathic α-helix를 형성하고 있는 것을 볼 수 있었다. 사람과 쥐의 SNAP-25 유전자는 88%, 생쥐와 쥐의 경 우는 97%의 homology를 보여 주고 있었다. 그리고 사람과 쥐의 ORF에서 염기서열은 94%, 생쥐와 쥐의 ORF에서 염기서열은 100%의 homology를 보여주고 있었으며 사람, 생쥐, 그리고 쥐 의 ORF에서 아미노산 서열은 100%의 homology를 보여 주고 있었다. SNAP-25 was first investigated as a neuron-specific protein preferentially expressed in CA3 pyramidal neurons of mouse hippocampus. It is a presynaptic plasma membrane protein in the nerve cell and plays an important role in the synaptic vesicle membrane docking and fusion pathway. We have recently isolated SNAP-25 cDNA from a rat brain cDNA library using a probe of Z2 cDNA. It consisted of 2,101 bp and an open reading frame (ORF) was identified between nucleotides (nt) 209 and 827. The AUG codon (nt 209∼211) was surrounded by CTACCATGG, which corresponded to the consensus sequence of ribosomal binding site. The ORF was terminated by TAA (nt 827∼829) to encode a polypeptide of 206 amino acid residues. The 3'-untranslated region contained two extensive stretches of repeated (CA)28 and (CA)19 at positions 925∼980 and 1645∼1682. It is noteworthy that cysteine residues were clustered in the span of amino acid residues 84∼91 : Cys-Gly-Leu-Cys-Val-Cys-Pro-Cys. Rat SNAP-25 showed 88% and 97% identity in nucleotide sequences to that of human and mouse, respectively. Amino acid sequence of rat SNAP-25 showed 100% identity to that of mouse and human SNAP-25.
조애리,동희진,서건호,조성범 한국식품과학회 2014 Food Science and Biotechnology Vol.23 No.2
A rapid and sensitive loop-mediated isothermalamplification (LAMP) assay was developed for detectingListeria monocytogenes prfA in milk. The inclusivity of 23L. monocytogenes and the exclusivity of 16 non-L. monocytogenes strains were both 100% in the assay. Thelimit of detection (LoD) of the LAMP assay in Listeriaenrichment broth (LEB) was 2.22 CFU/mL after 12 h and24 h of incubation. The LoDs of the LAMP assay in LEBwith artificially contaminated milk (LEB-M) incubated for12 h (2.22×101 CFU/mL) and 24 h (2.22 CFU/mL) werelower than those of the PCR and real-time PCR assays. Comparison of the LoDs in LEB with those in LEB-Mshowed that the LAMP assay was less influenced by themilk compounds than the real-time PCR assay. Our resultsindicate that the LAMP assay can be utilized as a potentialscreening tool for L. monocytogenes in milk.