• 제목/요약/키워드: Gene isolation

검색결과 863건 처리시간 0.059초

Molecular Cloning of the Superoxide Dismutase Gene from Orientia tsutsugamushi, the Causative Agent of Scrub Typhus

  • Koh, Young-Sang;Yun, Ji-Hyun;Kim, Se-Jae
    • Journal of Microbiology
    • /
    • 제40권2호
    • /
    • pp.151-155
    • /
    • 2002

  • A Superoxide Dismutase (SOD) gene from the obligate intracellular bacterium Orientia tsutsugamushi has been cloned by using the polymerase chain reaction with degenerate oligonucleotide primers corresponding to conserved regions of known SODs. Nucleotide sequencing revealed that the predicted amino acid sequence was significantly more homologous to known iron-containing SODs (FeSOD) than to manganese-containing SODs (MnSOD). Conserved regions in bacterial FeSOD could also be seen. Isolation of the oriential SOD gene may provide an opportunity to examine its role in the intracellular survival of this bacterium.

  • PDF KSCI

Isolation of Cold Acclimation-related Genes in Wheat Chromosome Substitution Line 5D

  • Chun, Jong Un;Jeong, In Ho
    • 한국육종학회지
    • /
    • 제40권3호
    • /
    • pp.234-242
    • /
    • 2008

  • To identify low temperature-induced genes of wheat chromosome substitution line 5D, suppression subtractive hybridization (SSH) was performed with mRNAs from leaf samples that treated with low temperature ($4^{\circ}C$). A cDNA library was constructed using mRNA isolated from wheat chromosome substitution line 5D leaves treated with low temperature ($4^{\circ}C$). The nucleotide and deduced amino acid sequences of the putative gene products were compared. wfr-9 and wfr-32 showed identity over 90% related to vernalization gene. Other two genes, wfr-77 and wfr-83 which is related to freezing-resistant gene have also identity over 90%. This result suggest that those genes may be transcribed into antifreeze proteins which are accumulated within leaf apoplasts, when wheat chromosome substitution line 5D is acclimated during low temperature treatment.

  • KSCI

TAR cloning 법에 의한 인간 및 마우스의 상동성 HPRT 유전자의 분리 (Isolation of Human and Mouse Orthologue HPRT Genes by Transformation-Associated Recombination (TAR) cloning)

  • 도은주;김재우;정정남;박인호;임선희
    • 생명과학회지
    • /
    • 제16권6호
    • /
    • pp.1036-1043
    • /
    • 2006

  • TAR (Transformation-Associated Recombination) cloning법은 복잡한 고등생물의 게놈으로부터 유전자나 특정 염색체 부위를 선별적 분리를 가능하게 한다. 이 방법은 목적으로 하는 염색체 부위의 주변에 존재하는 비교적 짧은 게놈 염기서열에 대한 정보를 필요로 한다. 이 기술은 출아효모의 spheroplasts 형질전환 동안 목적 유전자를 포함한 게놈 DNA와 그 유전자의 5' 또는 3' 말단 서열 (hook)을 포함하고 있는 TAR vector 사이에 일어나는 상동성 재조합에 의해 이루어진다. 본 연구에서는 TAR cloning 법을 상동성 유전자의 분리에 사용할 수 있는가를 조사하기 위해, 연간과 마우스 게놈의 HPRT 유전자를 선택하였다. 그 결과, 인간과 마우스의 게놈으로부터의 HPRT 유전자의 분리 빈도는 TAR vector로서 hHPRT hook 혹은 mHPRT hook을 사용한 경우에 거의 동일하게 나타났다. 또한 mHPRT 유전자의 gap 부분의 염기서열을 결정하여, 이 부분에 염기서열의 불안정의 요인이 되는 비정상적 특성을 발견하였다. 결론적으로 TAR cloning법을 이용하여 다른 이종 간의 게놈으로부터 상동성 유전자 즉 orthologue의 분리가 가능하였다. 더욱이 TAR cloning 시스템을 이용하여 고등동물 게놈 상에 남아있는 gap 부분을 메움으로서 고등동물의 모든 유전자들의 확인이 가속화될 수 있을 것으로 사료된다.

  • https://doi.org/10.5352/JLS.2006.16.6.1036 인용 PDF KSCI

Identification and Isolation of Differentially Expressed Gene in Response to Cold Stress in a Green Alga, Spirogyra varians (Zygnematales)

  • Han, Jong-Won;Yoon, Min-Chul;Lee, Key-Pyoung;Kim, Gwang-Hoon
    • ALGAE
    • /
    • 제22권2호
    • /
    • pp.131-139
    • /
    • 2007

  • The expression of genes responding to cold stress in a freshwater alga, Spirogyra varians, was studied by using differential expression gene (DEG) method. A gene strongly up-regulated in 4°C was isolated and designated as SVCR2 (Spirogyra varians cold regulated) gene. The cDNA encoding SVCR2 was cloned using λZAP cDNA library of Spirogyra varians. The deduced amino acid had a sequence similarity with trans-membrane protein in Arabidopsis thaliana (Q9M2D2, 52.7%). Northern blot analysis demonstrated that transcript level of SVCR2 increased about 10 fold under low temperature (4°C), compared with that cultured at warm (20°C) conditions. The expression of SVCR2 was also affected by light conditions. When the plants were exposed to high light (HL) (1200 μmol photon m–2 s–1), the expression of SVCR2 began within 2 hrs. This gene expression lasted for 4 hrs and decreased afterwards. Under the blue light (470 nm) condition, the expression of this gene was induced in same way as HL treatment, even under less than 100 μmol photon m–2 s–1. But red light (650 nm) and UV-A irradiation did not affect the expression of SVCR2.

  • https://doi.org/10.4490/ALGAE.2007.22.2.131 인용 PDF KSCI

Isolation and Characterization of Kasugamycin Biosynthetic Genes from Streptomyces kasugaensis KACC 20262

  • JO YOU-YOUNG;LIU JING;JIN YING-YU;YANG YOUNG-YELL;SUH JOO-WON
    • Journal of Microbiology and Biotechnology
    • /
    • 제15권3호
    • /
    • pp.491-496
    • /
    • 2005

  • The biosynthetic gene cluster for the aminoglycoside antibiotic kasugamycin was isolated and characterized from the kasugamycin producing strain, Streptomyces kasugaensis KACC 20262. By screening a fosmid library using kasA, the gene encoding aminotransferase, we isolated a 22 kb DNA fragment. The fragment contained seventeen complete open reading frames (ORFs); one of these ORFs, kasD, was identified as the gene for dNDP-glucose 4,6-dehydratase, which catalyzes the conversion of dNDP-glucose to 4-keto-6-deoxy-dNDP-glucose. The enzyme showed a broad spectrum of substrate specificity. In addition, ksR was overexpressed in E. coli BL21 and proved to be a self-resistance gene against kasugamycin. These findings suggest that the isolated gene cluster is highly likely responsible for the biosynthesis of kasugamycin.

  • PDF KSCI