• 제목/요약/키워드: Amino acid sequence

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Genomic Organization of Penicillium chrysogenum chs4, a Class III Chitin Synthase Gene

  • Park, Yoon-Dong;Lee, Myung-Sook;Kim, Ji-Hoon;Jun Namgung;Park, Bum-Chan;Bae, Kyung-Sook;Park, Hee-Moon
    • Journal of Microbiology
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    • 제38권4호
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    • pp.230-238
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    • 2000
  • Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

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G-Proteins Expressed in the Ocellus of the Hydromedusan, Spirocodon saltatrix.

  • Iwasa, Tatsuo;Shimazaki, Yumiko;Yamamoto, Masamichi;Ohtsu, Kohzoh
    • Journal of Photoscience
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    • 제9권2호
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    • pp.278-280
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    • 2002
  • We have cloned a hydromedusan opsin cDNA and showed that the deduced amino acid sequence of the cytoplasmic loop between helices 5 and 6 (loop 5-6) was clearly different from that reported so far. The amino acid sequence of the loop 5-6 is important on determination of the specificity for the coupled G- protein. To clarify which class of G-protein mediates the phototransduction system in the ocellus of the hydromedusan, we investigated G-proteins expressed in the ocellus. By PCR against the cDNA of the ocellus with primers designed according to the conserved amino acid sequence in G-protein a subunit, we obtained three kinds of cDNA fragments. Based on the sequence similarities, ttwo of them (JGI and JG3) were classified as $G_{i}$ and $G_{q}$, respectively. The other one (JG2) was a new subtype within $G_{*}$ class. Electron microscopic immunocytochemistry with the antiserum against the C-terminal sequence of $G_{q}$ or $G_{t}$ revealed the presence. of the both classes in the ocellus. The similarity of the C-terminal sequence of the JG2 with that of bovine $G_{t}$ suggests that the anti- $G_{t}$ antiserum would bind to JG2. These results suggest the possibility that the hydromedusan rhodopsin decides the specificity for the coupled G-protein by the other domain than the loop 5-6.oop 5-6.5-6.

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Complete Genome Sequence Analysis of Two Divergent Groups of Sweet potato chlorotic fleck virus Isolates Collected from Korea

  • Kwak, Hae-Ryun;Kim, Jaedeok;Kim, Mikyeong;Seo, Jang-Kyun;Kim, Jeong-Soo;Choi, Hong-Soo
    • The Plant Pathology Journal
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    • 제34권5호
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    • pp.451-457
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    • 2018
  • The Sweet potato chlorotic fleck virus (SPCFV), of the genus Carlavirus (family Betaflexiviridae), was first detected as one of several viruses infecting sweet potatoes (Ipomea batatas L.) in Korea. Out of 154 sweet potato samples collected in 2012 that were showing virus-like symptoms, 47 (31%) were infected with SPCFV, along with other viruses. The complete genome sequences of four SPCFV isolates were determined and analyzed using previously reported genome sequences. The complete genomes were found to contain 9,104-9,108 nucleotides, excluding the poly-A tail, containing six putative open reading frames (ORFs). Further, the SPCFV Korean isolates were divided into two groups (Group I and Group II) by phylogenetic analysis based on the complete nucleotide sequences; Group I and Group II had low nucleotide sequence identities of about 73%. For the first time, we determined the complete genome sequence for the Group II SPCFV isolates. The amino acid sequence identity in coat proteins (CP) between the two groups was over 90%, whereas the amino acid sequence identity in other proteins was less than 80%. In addition, SPCFV Korean isolates had a low amino acid sequence identity (61% CPs and 47% in the nucleotide-binding protein [NaBp] region) to that of Melon yellowing-associated virus (MYaV), a typical Carlavirus.

Cloning, Expression, and Nucleotide Sequencing of the Gene Encoding Glucose Permease of Phosphotransferase System from Brevibacterium ammoniagenes

  • Yoon, Ki-Hong;Yim, Hyouk;Jung, Kyung-Hwa
    • Journal of Microbiology and Biotechnology
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    • 제8권3호
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    • pp.214-221
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    • 1998
  • A Brevibacterium ammoniagenes gene coding for glucose/mannose-specific enzyme II ($EII^{Glc}$) of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing an Escherichia coli mutation affecting a ptsG gene, and the complete DNA nucleotide sequence was determined. The cloned gene was identified to be a ptsG, which enables the E. coli transportment to use glucose more efficiently than mannose as the sole carbon source in an M9 minimal medium. The ptsG gene of B. ammoniagenes consists of an open reading frame of 1,983 nucleotides putatively encoding a polypeptide of 661 amino acid residues and a TAA stop codon. The deduced amino acid sequence of the B. ammoniagenes $EII^{Glc}$ shows, at $46\%$, the highest degree of sequence similarity with the Corynebacterium glutamicum EII specific for both glucose and mannose. In addition, the $EII^{Glc}$ shares approximately $30\%$ sequence similarities with sucrose-specific and ${\beta}$-glucoside-specific EIIs of the several bacteria belonging to the glucose-PTS class. The 161-amino-acid C-terminal sequence of $EII^{Glc}$ is also similar to that of E. coli enzyme $IIA^{Glc}$, specific for glucose ($EIIA^{Glc}$). The B. ammoniagenes $EII^{Glc}$ consists of three domains; a hydrophobic region (EIIC) and two hydrophilic regions (EIIA, EIIB). The arrangement of structural domains, IIBCA, of the $EII^{Glc}$ is identical to those of EIIs specific for sucrose or ${\beta}$-glucoside. While the domain IIA was removed from the B. ammoniagenes $EII^{Glc}$ the remaining domains IIBC were found to restore the glucose and mannose-utilizing capacity of E. coli mutant lacking $EII^{Glc}$ activity with $EIIA^{Glc}$ of the E. coli mutant. $EII^{Glc}$ contains a histidine residue and a cysteine residue which are putative phosphorylation sites for the protein.

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Cloning and Sequence Analysis of Glyceraldehyde-3-Phosphate Dehydrogenase Gene in Yak

  • Li, Sheng-Wei;Jiang, Ming-Feng;Liu, Yong-Tao;Yang, Tu-Feng;Wang, Yong;Zhong, Jin-Cheng
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권11호
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    • pp.1673-1679
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    • 2008
  • In order to study the biological function of gapdh gene in yak, and prove whether the gapdh gene was a useful intra-reference gene that can be given an important role in molecular biology research of yak, the cDNA sequence encoding glyceraldehyde-3-phosphate dehydrogenase from yak was cloned by the RT-PCR method using gene specific PCR primers. The sequence results indicated that the cloned cDNA fragment (1,008 bp) contained a 1,002 bp open reading frame, encoding 333 amino acids (AAs) with a molecular mass of 35.753 kDa. The deduced amino acids sequence showed a high level of sequence identity to Bos Taurus (99.70%), Xenopus laevis (94.29%), Homo sapiens (97.01%), Mus musculus (97.90%) and Sus scrofa (98.20%). The expression of yak's gapdh gene in heart, spleen, kidney and brain tissues was also detected; the results showed that the gapdh gene was expressed in all these tissues. Further analysis of yak GAPDH amino acid sequence implied that it contained a complete glyceraldehyde-3-phosphate dehydrogenase active site (ASCTTNCL) which ranged from 148 to 155 amino acid residues. It also contained two conserved domains, a NAD binding domain in its N-terminal and a complete catalytic domain of sugar transport in its C-terminal. The phylogenetic analysis showed that yak and Bos taurus were the closest species. The prediction of secondary structures indicated that GAPDH of yak had a similar secondary structure to other isolated GAPDH. The results of this study suggested that the gapdh gene of yak was similar to other species and could be used as the intra-reference to analyze the expression of other genes in yak.

강아지 프로카이모신의 전 아미노산 서열 (The Complete Amino Acid Sequence of Newborn Dog Prochymosin)

  • Yoon, Joo-Ok;Kim, Hyun-Ku
    • 동아시아식생활학회지
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    • 제7권3호
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    • pp.289-300
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    • 1997
  • 생후 2주일 되는 강아지의 위에서 카이모신을 추출하고 이온교환 크로마토그래피로 정제하였다. 카이모신 아미노산 서열의 반은 아미노산 서열 분석법으로, 또 프로카이모신의 전아미노산 서열은 프로카이모신 cDNA의 염기서열로부터 결정하였다. 강아지 프로카이모신의 아미노산 서열은 송아지와는 79%, 돼지 펩신노진 A와는 54%의 상동성을 보였다. 프로펩티드의 크기와 활성효소의 N-말단 아미노산 잔기의 위치는 다른 프로카이모신과 같았다. 강아지 카이모신의 pH 3.2에서 단백질 분해활성의 최대 값은 돼지 펩신의 pH 2에서 값의 3-4% 밖에 되지 않았으나, 웅유활성은 송아지보다 훨씬 높았다. 강아지의 위 추출물에 대한 pH 5.2에서의 한천 젤 전기이동으로 프로카이모신과 카이모신에는 두 가지의 현저한 유전적 변이형이 존재함을 확인하였다. 두 변이형은 아미노산 조성, N-말단 서열, 그리고 효소성질에서 차이가 없었다. 송아지와 강아지 카이모신의 기질결합에 관여하는 아미노산 잔기는 다음과 같이 서로 달랐다(돼지 펩신의 서열번호로 표시함) : Ser12 Thr (S$_4$), Leu30 Val (S$_1$/S$_3$), His 74 Gln (S'$_2$), Val111 Ile (S$_1$/S$_3$), Lys220 Met (S$_4$). 강아지 카이모신의 단백질 분해활성이 낮은 것은 송아지의 Asp 303이 강아지에서는 Val303으로 바뀐 때문이라고 생각된다.

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Comparison of the Nucleotide Sequence of Cloned Osteopontin from Hanwoo and Holstein

  • Lee, Tae Young;Ju, Sung Kyu;Nam, Myoung Soo
    • 한국축산식품학회지
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    • 제33권3호
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    • pp.331-334
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    • 2013
  • Osteopontin (OPN) is a secreted phosphorylated glycoprotein. It has an important role in multiple biological processes including cell survival, bone remodeling, inhibition of ectopic calcification, as well as, is thought to have potential immune modulation activities. In this work, we isolated and characterized a full-length open reading frame (ORF) of Korean native cow's OPN from Korean native cow's (Hanwoo) kidney, and successfully cloned firstly on Hanwoo's OPN. The sequencing results indicated that the isolated cDNA was 1190 bp in length containing a complete ORF of 837 bp. It encoded a precursor protein Hanwoo's OPN consisting of 278 amino acids with a signal peptide of 16 amino acids. Amino acid homology was found to be 99.3% as compared to the corresponding sequences of Holstein bone marrow OPN. Hanwoo's kidney OPN and Holstein bone marrow OPN are different only in two amino acid residues 42 and 56, amino acid residue 42 is Thr (T) ${\leftrightarrow}$ Ile (I), and amino acid residue 56 is Ala (A) ${\leftrightarrow}$ Thr (T) respectively. These results from the present work would be helpful to elucidate the biological function of Hanwoo's OPN and provided a foundation for further insight into role of Hanwoo's OPN.

Screening and Purification of a Novel Transaminase Catalyzing the Transamination of Aryl ${\beta}-Amino$ Acid from Mesorhizobium sp. LUK

  • Kim, Ju-Han;Kyung, Do-Hyun;Yun, Hyung-Don;Cho, Byung-Kwan;Kim, Byung-Gee
    • Journal of Microbiology and Biotechnology
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    • 제16권11호
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    • pp.1832-1836
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    • 2006
  • Mesorhizobium sp. LUK, which utilizes 3-amino-3-phenylpropionic acid as the sole source of nitrogen with high enantioselectivity (E(S)>100), was isolated using enrichment culture. The enzyme involved in the utilization of (S)-3-amino-3-phenylpropionic acid was confirmed to be a transaminase and was purified by 235-folds with a specific activity of 0.72 U/mg. The molecular weight of the purified protein was ca. 47 kDa and the active enzyme was determined as a dimer on gel filtration chromatography. The N-terminal sequence was obtained from the purified protein. Spontaneous decarboxylation of produced ${\beta}-keto$ acids was observed during the chiral resolution of 3-amino-3-phenylpropionic acid.

버섯의 유리당, 당알콜, 아미노산 및 무기질의 조성에 관한 연구 (Studies on Free-Sugars, Sugaralcohols, Amino acids and Mineral Contents in Edible Mushrooms.)

  • 허윤행
    • 환경위생공학
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    • 제4권2호
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    • pp.27-32
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    • 1989
  • In orde to investigate free sugar, sugar alcohol, amino acid and mineral contents of edible mushrooms, hentinus edodes, Auricula-Jude and Coriolus Versicolor were analyzed. 1. In each of the three mushrooms, glucose, fructose, xylose, trehalose, mannose and mannitol were identified and in the hentinus edodes and Auricula-jude, the major free sugar was trehalose, respectively and increasing sequence in amount of free sugars were mannitol, mannose, fructose, glucose, on the other hand that lowest amount was Xylose. The relatively richest were glucose, fructose, trehalose, mannose, on the other hand lower amount of sample, Coriolus Versicolor were mannitol and Xylose. 2. In each of the three samples, essential amino acids were high amounts, especially good taste component, glutamic acid was higher. 3. The mineral p Contents$(925\~115mg)$ were highest and Mg was higher amount, in hentinus edodes and Auricula-jude, K, Na, Ca, Cu, Zn, etc were identified, In Coriolus versicolor, K content was highest, on the other hand, Sequence of mineral amount was Ca, Fe and Na.

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Biological activity of peptides purified from fish skin hydrolysates

  • Abuine, Racheal;Rathnayake, Anuruddhika Udayangani;Byun, Hee-Guk
    • Fisheries and Aquatic Sciences
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    • 제22권5호
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    • pp.10.1-10.14
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    • 2019
  • Fish skin waste accounts for part of the solid waste generated from seafood processing. Utilization of fish skin by bioconversion into high-grade products would potentially reduce pollution and economic cost associated with treating fish processing waste. Fish skin is an abundant supply of gelatin and collagen which can be hydrolyzed to produce bioactive peptides of 2-20 amino acid sequences. Bioactivity of peptides purified from fish skin includes a range of activities such as antihypertensive, anti-oxidative, antimicrobial, neuroprotection, antihyperglycemic, and anti-aging. Fish skin acts as a physical barrier and chemical barrier through antimicrobial peptide innate immune action and other functional peptides. Small peptides have been demonstrated to possess biological activities which are based on their amino acid composition and sequence. Fish skin-derived peptides contain a high content of hydrophobic amino acids which contribute to the antioxidant and angiotensin-converting enzyme inhibitory activity. The peptide-specific composition and sequence discussed in this review can be potentially utilized in the development of pharmaceutical and nutraceutical products.