• Title/Summary/Keyword: Multigene analysis

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Molecular Characterization of a Chinese Cabbage cDNA Encoding Thioredoxin-h that is Predominantly Expressed in Flowers

  • Lee, Seung-Sik;Lee, Kyun-Oh;Jung, Bae-Gyo;Chi, Yong-Hun;Yoo, Ji-Young;Lee, Ji-Yeun;Lee, Jung-Ro;Park, Soo-Kwon;Kang, Soon-Suk;Jang, Ho-Hee;Lee, Sang-Yeol
    • BMB Reports
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    • v.34 no.4
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    • pp.334-341
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    • 2001
  • Even though three isotypes of thioredoxins (-f, -m and -h types) have been identified in a variety of plant cells, there are only a few reports on thioredoxin-h that were recently identified. In this study, a cDNA encoding a h-type of thioredoxin was isolated from a cDNA library of Chinese cabbage, and named here CTrx-h. An open reading frame of the gene contained a polypeptide of 133 amino acids with a conserved active center, WCGPC, which appeared in all of the thioredoxin proteins. A deduced amino acid sequence of the CTrx-h showed the highest sequence identity with those of Arabidopsis thioredoxin-h2 (75.2%) and thioredoxin-h5 (46.6%) proteins, but it shared a low sequence homology to other isotypes of plant thioredoxinm and thioredoxin-f. The CTrx-h protein that is expressed in E. coli represented not only an insulin reduction activity, but also electron transferring activity from NADPH to thioredoxin-dependent peroxidase. A genomic Southern blot analysis using the cDNA insert of CTrx-h revealed that the gene consisted of a small multigene family in Chinese cabbage genome. On the contrary to other thioredoxin-h proteins that were widely distributed in most tissues of the plant, the CTrx-h was predominantly expressed in flowers. The expression was very low in other tissues. The data of the Northern blot analysis suggests that the CTrx-h may have other functions in flower development or differentiation, in addition to its defensive role.

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Expression Analysis of OsCPK11 by ND0001 oscpk11 Mutants of Oryza sativa L. under Salt, Cold and Drought Stress Conditions (염분, 저온 및 가뭄 스트레스 조건에서 벼 ND0001 oscpk11 돌연변이체의 OsCPK11 발현 분석)

  • Kim, Hyeon-Mi;Kim, Sung-Ha
    • Journal of Life Science
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    • v.31 no.2
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    • pp.115-125
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    • 2021
  • Calcium-dependent protein kinases (CDPKs) are known to be involved in regulating plant responses to abiotic stresses such as salinity, cold temperature and dehydration,. Although CDPKs constitute a large multigene family consisting of 31 genes in rice, only a few rice CDPKs' functions have been identified. Therefore, in order to elucidate the functions of OsCPK11 in rice, this study was intended to focus on the expression pattern analysis of OsCPK11 in wild type and ND0001 oscpk11 mutant plants under these abiotic stresses. For the salt, cold and drought stress treatment, seedlings were exposed to 200 mM NaCl, 4℃ and 20% PEG 6,000, respectively. RT-PCR and quantitative real-time PCR were performed to determine the expression patterns of OsCPK11 in wild type and ND0001 mutant plants. RT-PCR results showed that OsCPK11 transcripts in the wild type and heterozygous mutant were detected, but not in the homozygous mutant. Real-time PCR results showed that relative expression of OsCPK11 of wild type plants was increased and reached to the highest level at 24 hr, at 6 hr and at 24 hr under salt, cold and drought stress conditions, respectively. Relative expression of OsCPK11 of ND0001 homozygous plant was significantly reduced compared to that of wild type. These results suggested that oscpk11 homozygous mutant knocks out OsCPK11 and OsCPK11 might be involved in salt, cold and drought stress signaling by regulating its gene expression.

Isolation and Characterization of ACC Synthase Gene Family in Mung Bean (Vigna radiata L.): Differential Expression of the Three ACC Synthase enes in Response to Auxin and Brassinosteroid

  • Sunjoo Joo;Kim, Woo-Taek
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.61-71
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    • 2000
  • By screening a cDNA library of auxin-treated mung bean (Vigna radiata L.) hypocotyls, we have isolated two full-length cDNA clones, pVR-ACS6 and pVR-ACS7, for 1-aminocyclopropane-1-carboxylate (ACC) synthase, the rate-limiting enzyme in the ethylene biosynthetic pathway. While PVR-ACS6 corresponds to the previously identified PCR fragment pMBA1, pVR-ACS7 is a new cDNA clone. A comparison of deduced amino acid sequences among auxin-induced ACC synthases reveal that these enzymes share a high degree of homology (65-75%) to VR-ACS6 and VR-ACS7 polypeptides, but only about 50% to VR-ACS1 polypeptide. ACS6 and ACS7 are specifically induced by auxin, while ACS1 is induced by cycloheximide, and to lesser extent by excision and auxin treatment. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, whereas W-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the $\beta$-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tobacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR.

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cDNA Cloning and Polymorphism of the Porcine Carbonic Anhydrase III (CA3) Gene

  • Wu, J.;Deng, Changyan;Xiong, Y.Z.;Zhou, D.H.;Lei, M.G.;Zuo, B.;Li, F.E.;Wang, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.3
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    • pp.324-328
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    • 2006
  • Carbonic anhydrase III (CA3) is a member of a multigene family that encode carbonic anhydrase isozymes. In this study, a complete coding sequence of the pig CA3 gene which encodes a 260 amino-acid protein was determined. The amino acid comparison showed high sequence similarities with previously identified human (86.5%) CA3 gene and mouse (91.5%) Car3 gene. The partial genomic DNA sequences were also investigated. The length of intron 1 was 727 bp. Comparative sequencing of three pig breeds revealed that there was a T${\rightarrow}$C substitution at position 363 within intron 1. The substitution was situated within a NcoI recognition site and was developed as a PCR-restriction fragment length polymorphism (RFLP) marker for further use in population variation investigations and association analysis. Two alleles (A and B) were identified, and 617 bp fragments were observed for the AA genotype and 236 bp and 381 bp fragments for the BB genotype. The polymorphism of CA3 was detected in 8 pig breeds. Allele B was predominant in the Western pig breeds. In addition, association studies of the CA3 polymorphism with carcass traits in 140 $Yorkshire{\times}Meishan$ $F_2$ offspring showed that the NcoI PCR- RFLP genotype may be associated with variation in several carcass traits of interest for pig breeding. Allele B was associated with increases in lean meat percentage, loin eye height and loin eye area. Statistically significant association with backfat thickness was also found; pigs with the AB genotype had much less backfat thickness than AA or BB genotypes.

Production of stable chloroplast-transformed plants in potato (Solanum tuberosum L.) (안정적 감자 엽록체 형질전환 식물체 생산)

  • Min, Sung-Ran;Jeong, Won-Joong;Park, Ji-Hyun;Lyu, Jae-Il;Lee, Jeong-Hee;Oh, Kwang-Hoon;Chung, Hwa-Jee;Liu, Jang-R.
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.42-48
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    • 2011
  • Chloroplast genetic engineering of higher plants offers several unique advantages compared with nuclear genome transformation, such as high levels of transgene expression, a lack of position effect due to site-specific transgene integration by homologous recombination, multigene engineering in a single transformation event and reducing risks of gene flow via pollen due to maternal inheritance. We established a reproducible chloroplast transformation system of potato using a tobacco specific plastid transformation vector, pCtVG (trnI-Prrn-aadA-mgfp-TpsbA-trnA). Stable transgene integration into chloroplast genomes and the homoplasmic state of the transgenome were confirmed by PCR and Southern blot analyses. Northern, immunoblot analysis, and GFP fluorescence imaging revealed high expression and accumulation of GFP in the plastids of potato leaves. This system would provide new opportunities for genetic improvement and mass production of value added foreign proteins in this crop.

Phylogeny of Desmodesmus (Scenedesmaceae, Chlorophyceae) in Korea based on multigene data analysis (다유전자 분석을 통한 한국산 녹조류 Desmodesmus속의 계통)

  • Yeong Chae Yoo;Nam-Ju Lee;Ga Yeong Jeon;Ok-Min Lee;Eun Chan Yang
    • Korean Journal of Environmental Biology
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    • v.41 no.4
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    • pp.345-363
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    • 2023
  • The genus Desmodesmus (Chodat) S.S. An, T. Friedl & E. Hegewald is ubiquitous in freshwater ecosystems, such as rivers, ponds, and wetlands. The actual species diversity and distribution of the genus is unknown because of morphological plasticity affected by habitats. Currently, 38 Desmodesmus species have been reported in Korea most of which transferred from the genus Scenedesmus recently, however, no phylogenetic relationships have been studied yet. Despite the challenges in analyzing relationships among Desmodesmus species through the morphology, ecology, and original description, this study focused on examining species-level relationships using the FBCC culture strains isolated from Korea. A total of 299 sequences (66 of 18S rRNA, 47 of atpB, 67 of petA, 52 of rbcL, and 67 of tufA) were newly determined and used for phylogenetic analysis. Four plastid genes tend to have higher variation than 18S rRNA in the variable sites and P-distance. From the combined phylogeny, the Desmodesmus included six clades such as Clade-1: D. pseudoserratus and D. serratus, Clade-2: D. communis, D. dispar, D. maximus, D. pannonicus, unidentified Desmodesmus sp., Clade-3: D. bicaudatus and D. intermedius, Clade-4: D. microspina, D. multivariablis, D. pleiomorphus, D. subspicatus, Clade-5: D. abundans, D. kissii, and D. spinosus, and Clade-6: D. armatus, D. armatus var. longispina, D. opoliensis, unidentified Desmodesmus spp. The new sequence data from FBCC strains will be used to identify species and study the molecular ecology of scenedesmacean green algae in freshwater ecosystems. The phylogenetic information from this study will expand our understanding of Desmodesmus species diversity in Korea.