• Title/Summary/Keyword: chain length determinant

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Identification and Functional Analysis of the Chain Length Determinant Gene ste8 Involved in the Biosynthesis of Ebosin by Streptomyces sp. 139

  • Yang, Zhang;Li, Xiaohua;Qi, Xiaoqaing;Shan, Junjie;Jiang, Rong;Guo, Lianhong;Zhang, Ren;Li, Yuan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1500-1508
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    • 2013
  • Ebosin, a novel exopolysaccharide produced by Streptomyces sp. 139, has obvious antirheumatic arthritis activity in vivo, and its biosynthesis gene cluster (ste), consisting of 27 open reading frames, has been identified. This paper reports our study of the gene functionality of ste8, the predicted protein product of which is homologous to some bacterial chain length determinant Wzz proteins. For characterization of Ste8, ste8 was cloned and expressed in the mutant strain E. coli 086:H2 (${\Delta}wzz$). The functional complementation of wzz by ste8 was demonstrated by the restoration of wild-type lipopolysaccharide biosynthesis and increased levels of serum resistance of E. coli 086:H2 (${\Delta}wzz$) (pET30a-ste8). To examine the function of ste8 in ebosin biosynthesis, the gene was knocked out with a double crossover via homologous recombination. The molecular weight of the ebosin derivative EPS-8m produced by the mutant Streptomyces sp. 139 ($ste8^-$) was much lower than that of ebosin, and the binding activity of EPS-8m for IL-1R decreased significantly compared with ebosin. These results demonstrate that ste8 encodes a chain length determinant (Wzz) that functions in ebosin biosynthesis.

Symptom Determinant as RNA3 of Lily Isolates of Cucumber mosaic virus on Zucchini Squash

  • Cho, Seung-Kook;Ahn, Hong-Il;Kim, Min-Jea;Choi, Jang-Kyung;Ryu, Ki-Hyun
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.212-219
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    • 2004
  • Three isolates of Cucumber mosaic virus (CMV) from lily plants showing mosaic and distortion symptoms were detected by reverse-transcriptase polymerase chain reaction (RT-PCR) using primers specific to Cucumovirus genus namely, LK-CMV, LK4-CMV, and LKS-CMV. Restriction enzymes patterns of the RT-PCR products revealed that the lily isolates belonged to subgroup IA of CMV. In terms of biological properties, the lily isolates have highly similar but distinct pathogenicity as reported in other lily strains and ordinary strains of CMV. To characterize the molecular properties, cDNAs containing coat protein (CP) gene and 3' non-coding region (NCR) of RNA3 for the isolates were cloned and their nucleotide sequences were determined. The CP similarity (218 amino acids) was highly homologous (>97%) with that of subgroup I CMV strains. However, an additional 20-nulcleotide long segment was only present in 3' NCR of lily isolates, which form an additional stem-loop RNA structure. By using chimeric construct exchange cDNA containing 3'NCR of LK-CMV into the full-length cDNA clone of RNA3 of Fny-CMV, this additional segment may prove to be significant in the identification and fitness of the virus in lily plants. The pathology of zucchini squash infected by F1F2L3-CMV, a pseudorecombinant virus was showed to change drastically the severe mosaic and stunting symptom into a mild chlorotic spot on systemic leave, compared with Fny-CMV. To delimit the sequence of RNA3 affected the pathology, various RNA3 chimeras were constructed between two strains of CMV. The symptom determinants of F1F2L3-CMV were mapped to the positions amino acid 234, 239, and 250 in 3a movement protein (MP). RNA3 chimeras changed the sequences encoding three amino acids were resulted in alteration of systemic symptom.

Physicochemical Characteristics of Starches in Rice Cultivars of Diverse Amylose Contents

  • Yoon, Mi-Ra;Chun, A-Reum;Oh, Sea-Kwan;Hong, Ha-Cheol;Choi, Im-Soo;Lee, Jeong-Heui;Cho, Young-Chan;Kim, Yeon-Gyu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.3
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    • pp.226-232
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    • 2012
  • Through the sampling four rice cultivars with differing amylose contents, the relationship between the structural and gelatinization properties of endosperm starches was analyzed. These rice varieties exhibited different chain length distribution ratio within the amylopectin cluster as well as varing amylose levels. The proportion of amylopectin short chains of in Goami cutlivars was higher than the other varieties, whereas the Goami 2 which shows amylose extender mutant properties in the endosperm showed the highest proportion of long chains. In X-ray diffraction analysis of rice starches, the Goami 2 variety displayed a B-type pattern whereas the other varieties were all A-type. Among the cultivars with high and normal rice starch levels, those with the higher amylose contents showed distinctly lower swelling. Goami 2 rice was found to have the highest onset and peak gelatinization temperature from the differential scanning calorimetry results. The four rice cultivars under analysis also showed different rates of hydrolysis by amyloglucosidase. These findings suggest that the composition and chemical structure of the starch content is a major determinant of both the gelatinization and functional properties of rice.