• 제목/요약/키워드: ITS gene sequence

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Molecular cloning, expression and characterization of a novel feruloyl esterase enzyme from the symbionts of termite (Coptotermes formosanus) gut

  • Chandrasekharaiah, Matam;Thulasi, Appoothy;Bagath, M.;Kumar, Duvvuri Prasanna;Santosh, Sunil Singh;Palanivel, Chenniappan;Jose, Vazhakkala Lyju;Sampath, K.T.
    • BMB Reports
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    • 제44권1호
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    • pp.52-57
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    • 2011
  • Termites play an important role in the degradation of dead plant materials and have acquired endogenous and symbiotic cellulose digestion capabilities. The feruloyl esterase enzyme (FAE) gene amplified from the metagenomic DNA of Coptotermes formosanus gut was cloned in the TA cloning vector and subcloned into a pET32a expression vector. The Ft3-7 gene has 84% sequence identity with Clostridium saccharolyticum and shows amino acid sequence identity with predicted xylanase/chitin deacetylase and endo-1,4-beta-xylanase. The sequence analysis reveals that probably Ft3-7 could be a new gene and that its molecular mass was 18.5 kDa. The activity of the recombinant enzyme (Ft3-7) produced in Escherichia coli (E.coli) was 21.4 U with substrate ethyl ferulate and its specific activity was 24.6 U/mg protein. The optimum pH and temperature for enzyme activity were 7.0 and $37^{\circ}C$, respectively. The substrate utilization preferences and sequence similarity of the Ft3-7 place it in the type-D sub-class of FAE.

Bacillus subtilis WL-8의 Mannanase 유전자 클로닝과 특성분석 (Cloning and Characterization of Mannanase Gene from Bacillus subtilis WL-8)

  • 윤기홍
    • 미생물학회지
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    • 제46권2호
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    • pp.207-212
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    • 2010
  • 전통 발효식품인 된장으로부터 mannanase의 생산균으로 분리된 WL-8 균주는 형태적 특성, 생화학적 성질 및 16S rRNA의 염기서열에 근거하여 Bacillus subtilis로 동정되었다. B. subtilis WL-8은 locust bean gum 보다는 밀기울이 첨가된 LB 배지에서 mannanase 생산성이 높았으며, 24시간 배양하였을 때 약 20 U/ml에 이르렀다. 분리균 WL-8의 mannanase 유전자를 클로닝하여 그 염기서열을 결정한 결과 mannanase 유전자는 362 아미노산으로 구성된 단백질을 코드하며 1,086 뉴클레오티드로 이루어졌다. 아미노산 잔기배열을 분석한 결과 WL-8의 mannanase는 GH family 26에 속하며 B. subtilis의 mannanases와 매우 상동성이 높았다. B. subtilis WL-8의 mannanase 유전자를 함유한 재조합 대장균의 배양상등액과 균체파쇄상등액을 사용하여 반응특성을 조사한 결과 pH 5.5와 $60^{\circ}C$에서 최대 반응활성을 보였으며, $60^{\circ}C$보다 높은 온도에서 배양상등액보다는 균체파쇄상등액에 존재하는 mannanase가 더 높은 활성을 보였다.

Sequence Analysis of the Coat Protein Gene of a Korean Isolate of Iris Severe Mosaic Potyvirus from Iris Plant

  • Park, Won-Mok;Lee, Sang-Seon;Park, Sun-Hee;Ju;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • 제16권1호
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    • pp.36-42
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    • 2000
  • The coat protein gene of iris severe mosaic potyvirus, which was isolated in Korea, ISMV-K, from iris plant was cloned and its nucleotide sequence was determined. The coat protein of the virus contained 252 amino acid residues, including five potential N-glyxosylation site motifs. The coat protein of ISMV-K has 99.1% and 98.4% sequence identities with those of the Netherlands isolate of ISMV (ISMV-Ne) form crocus for the nucleotide and amino acids, respectively. The coat protein of ISMV-K has 50.4% to 60.3% nucleotide sequence identities and 47.3% to 55.7% amino acid identities with those of other 21 potyviruses, indicating ISMV to be a distinct species of the genus. The coat protein of ISMV-K was closely related with bean yellow mosaic virus and clover yellow vein virus in the phylogenetic tree analysis among the potyviruses analyzed. ISMV was easily and reliably detected from virus-infected iris leaves by RT-PCR with a set of the virus-specific primers.

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Nucleotide Sequence of Rat Transketolase and Liver-Specific Pretranslational Activation During Postnatal Development

  • Kim, Sung-Min F.;Kim, Byung-Moon;Jeng, Jingjau;Soh, Yun-Jo;Bak, Choong-Il;Huh, Jae-Wook;Song, Byoung-J.
    • BMB Reports
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    • 제29권2호
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    • pp.146-150
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    • 1996
  • A 2.1 kb cDNA clone for rat transketolase was isolated from rat liver ${\lambda}gt11$ cDNA library and its sequence was determined. The predicted rat transketolase (655 amino acids with $M_r$ 71,186) is highly similar (92%) to that of the human enzyme except that it contains an extra 32 amino acids at its N-terminus. Although it is less similar (<27%) to transketolases from non-mammalian species, the functional motifs such as the catalytic sites and thiamine binding domain are well conserved in the rat enzyme. Southern blot analysis of genomic DNA verified that transketolase appears to be derived from a single gene. Immunoblot and Northern blot analyses suggested that hepatic transketolase was activated pretranslationally by a 2.1-fold while little change was observed in brain enzyme, indicating a tissue-specific pretranslational activation during postnatal development.

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Bridging a Gap between DNA sequences and expression patterns of genes

  • Morishita, Shinichi
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2000년도 International Symposium on Bioinformatics
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    • pp.69-70
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    • 2000
  • The completion of sequencing human genome would motivate us to map millions of human cDNAs onto the unique ruler "genome sequence", in order to identify the exact address of each cDNA together with its exons, its promoter region, and its alternative splicing patterns. The expression patterns of some cDNAs could therefore be associated with these precise gene addresses, which further accelerate studies on mining correlations between motifs of promoters and expressions of genes in tissues. Towards the realization of this goal, we have developed a time-and-space efficient software named SQUALL that is able to map one cDNA sequence of length a few thousand onto a long genome sequence of length thirty million in a couple of minutes on average. Using SQUALL, we have mapped twenty thousand of our Bodymap (http://bodymap.ims.u-tokyo.ac.jp) cDNAs onto the genome sequences of Chr.21st and 22nd. In this talk, I will report the status of this ongoing project.

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Morphological and Multigene Sequence Characteristics of Talaromyces variabilis Isolated from Soil in Korea

  • Adhikari, Mahesh;Kim, Sang Woo;Lee, Hyang Burm;Lee, Youn Su
    • 한국균학회지
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    • 제49권1호
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    • pp.11-19
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    • 2021
  • In 2017, Talaromyces variabilis was isolated during a survey of fungal diversity in field soils in Korea. This isolate was described based on its morphological and molecular characteristics and it was identified molecularly using the partial 18S-ITS1-5.8S-ITS2-28S rDNA region and calmodulin (CaM)-encoding gene sequence data. Thus, this study reported morphological and multigene sequence characterization of T. variabilis.

Molecular Analysis of the 3'-Terminal Region of Lily Latent Carlavirus from Lilium lancitoium

  • Ryu, Ji-Hwan;Park, Hye-Won;Park, Won-Mok;Lee, Se-Yong;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • 제16권4호
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    • pp.231-235
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    • 2000
  • The 3,000 nucleotides of 3'-terminal region of the genomic RNA of a new isolate of carlavirus from a Korean native lily (Lilum lancitoium) was cloned and its nucleotide sequences were determined. The coat protein (CP) gene of the virus showed 72.0% to 72.8% nucleotide sequence identities and 86.9% to 88.0% amino acid sequence identities with those of the four strains (two Korean, one Dutch, and one Japanese isolates) of lily symptomless virus (LSV). Interestingly, different amino acid sequences between the new isolate and LSV strains were located at the N-terminal region of the CP. Pairwise amino acid sequence comparison of the CP gene revealed sequence identities of 22.0% to 71.1% between the virus and other 9 carlavirus species. The 25 kDa and 12 kDa proteins genes of the virus share 30.7% to 76.3% and 31.1% to 85.8% amino acid sequence identities, respectively, with those of 8 other carlaviruses. The 16 kDa protein gene of the virus shares 16.7% to 72.9% amino acid sequence identities with that of 9 other carlaviruses. These data indicate that the virus, designated as lily latent virus (LiLV), is a distinct of the Carlavirus genus and distinguished from the known strains of LSV.

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Cloning and Expression of the Duck Leptin Gene and the Effect of Leptin on Food Intake and Fatty Deposition in Mice

  • Dai, Han Chuan;Long, Liang Qi;Zhang, Xiao Wei;Zhang, Wei Min;Wu, Xiao Xiong
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권6호
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    • pp.850-855
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    • 2007
  • Leptin is the adipocyte-specific product of the obese gene and plays a major role in food intake and energy metabolism. Leptin research was mainly focused on mammalian species, but understanding of leptin and its function in poultry is very poor. In this study, the duck leptin gene was amplified using the reverse transcription-polymerase chain reaction (RT-PCR) from duck liver RNA. The cDNA fragment was inserted into the pET-28a expression vector, and the resulting plasmid was expressed in Escherichia coli BL21 (DE3). Experimental mice were given an intraperitoneal injection of 10 mg/kg leptin dissolved in phosphate buffered saline (PBS), while the control mice were injected with PBS. The effect of leptin on food intake, body weight and fatty deposition in mice was detected. Sequence analysis revealed that duck leptin had a length of 438 nucleotides which encoded a peptide with 146 amino acid residues. The sequence shares highly homology to other animals. The coding sequence of duck leptin was 84 and 86% identical to human and pig leptin nucleotides sequence. Highest identity was with the rat coding sequence (95%). The identity of the amino acid sequence was 84, 82 and 96% respectively compared to that of the human, pig and rat. Results of SDS-PAGE analysis indicated that a fusion protein was specifically expressed in E. coli BL21 (DE3). The purified product was found to be biologically active during tests. Continuous administration of recombinant duck leptin inhibited food intake. Despite the decrease of food intake, leptin significantly induced body weight and fatty deposition. These changes were accompanied by a significant down-secretion of plasma glucose, cholesterol, triglyceride and insulin levels in mice. The observations provide evidence for an inhibitory effect of leptin in the regulation of food intake and for a potential role of duck leptin in the regulation of lipogenesis.

Identification of a Third Haplotype of the Sequence Linked to the Restorer-of-fertility (Rf) Gene and Its Implications for Male-Sterility Phenotypes in Peppers (Capsicum annuum L.)

  • Min, Woong-ki;Lim, Heerae;Lee, Young-Pyo;Sung, Soon-Kee;Kim, Byung-Dong;Kim, Sunggil
    • Molecules and Cells
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    • 제25권1호
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    • pp.20-29
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    • 2008
  • Cytoplasmic male sterility (CMS), one of the most important traits in crop breeding, has been used for commercial seed production by $F_1$ hybrid cultivars of pepper (Capsicum annuum L.). To develop reliable molecular markers for allelic selection of the Restorer-of-fertility (Rf) gene, which is known to be a major determinant of pollen fertility restoration in peppers, a sequence of approximately 10 kb flanking an RAPD fragment closely linked to the Rf locus was obtained by genome walking. A homology search revealed that this sequence contained an LTR retrotransposon and a non-LTR LINE-like retrotransposon. Sequencing of this Rf-linked region to search for polymorphisms between a dominant and recessive allele revealed 98% nucleotide sequence identity between them. A third polymorphic haplotype of the Rf-linked sequence, which has 94-96% nucleotide sequence identity with the two previously isolated haplotypes, was identified among a large number of breeding lines. Utilizing polymorphic sequences in the haplotypes, PCR markers were developed for selection of particular haplotypes and used to examine the distribution of the haplotypes in diverse breeding lines, cultivars, and C. annuum germplasms. Surprisingly, the third haplotype was the predominant type in C. annuum germplasms, while its frequency in $F_1$ hybrid cultivars was relatively low. Meanwhile, analysis of breeding lines whose Rf allele genotypes and male-sterility phenotypes were already known revealed that the third haplotype was mainly present in exotic breeding lines that cause unstable male-sterility when combined with sterile cytoplasms.