• Title/Summary/Keyword: Isoforms

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Isolation of a cDNA Encoding a Chloroplast Triosephosphate Isomerase from Strawberry

  • Kim, In-Jung;Lee, Byung-Hyun;Jinki Jo;Chung, Won-Il
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.115-121
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    • 2000
  • A cDNA clone encoding chloroplast triosephosphate isomerase (TPI-cp) was isolated from strawberry fruit cDNA library. Sequence analyses indicated that the cDNA contains an open reading frame of 314 amino acids (33.5 kDa) composed of a transit peptide (59 amino acids) in amino terminal region and mature protein (255 amino acids). The existence of transit peptide in the deduced amino acid sequence implies that it encodes a chloroplast isoform. The protein sequence is more similar to other plant chloroplast isoforms than cytosolic isoforms. RNA blot analysis indicated that its expression is ubiquitous in examined five tissues, flowers, leaves, petioles, roots and fruits, and shows differential pattern according to fruit ripening. Genomic DNA blot analysis showed that TPI-cp is encoded by multiple genes in strawberry. Through sequence comparison and phylogenetic tree construction, TPI-cp is distinctively grouped into dicot and chloroplast isoforms.

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Expression of Neuregulins and Their Receptors During the Differentiation of Rat Hippocampal HiB5 Cells

  • Kwon, Hyockman
    • Animal cells and systems
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    • v.5 no.3
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    • pp.247-251
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    • 2001
  • Differentiating HiB5 cells, a rat hippocampal cell line, expressed neuregulins and showed constitutive activation of a neuregulin receptor, ErbB2, suggesting development of a neuregulin autocrine loop. RT-PCR analyses indicated that HiB5 cells produced SMDF and NDF, but not GGF, during the differentiation. None of neuregulin isoforms were detected in proliferating HiB5 cells. The neuregulins in HiBS cells, at least in part, are the $\beta$-isoforms of which the most of neuronal neuregulin isoforms are. The expression of SMDF and NDF was enhanced by PDGF and bFGF that promote cell survival and differentiation, suggesting a close relationship between the synthesis of neuregulins and the differentiation process. HiB5 cells have ErbB2 and ErbB4, but not ErbB3 receptors. Constitutive tyrosine phosphorylation of ErbB2 was detected in HiB5 cells that had not been exposed to exogenous GGF.

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Measuring of $Na^+-Ca^{2+}$ Exchange Current in Cerebellar Purkinje Cells

  • Cho, Chung-Hyun;Daejong Jeon;Shin, Hee-Sup;Lee, Chin O.
    • Proceedings of the Korean Biophysical Society Conference
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    • 2002.06b
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    • pp.37-37
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    • 2002
  • The Na$^{+}$-Ca$^{2+}$ exchanger (NCX) is known to playa critical role in the regulation of intracellular $Ca^{2+}$ in many tissues and cells. Three isoforms have been cloned (NCXl, NCX2, NCX3). Among the isoforms, NCX2 and NCX3 are expressed at high levels in brain and in a few other tissues. But the differential properties of the isoforms are not yet clearly established.(omitted)

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cDNA Cloning and Tissue Distribution of Two Parvalbumin Isoforms from the Hermaphrodite fish Rivulus marmoratus(Cyprinodontiformes, Rivulidae)

  • Lee, Jae-Seong;Lee, Young-Mi;Jung, Sang-Oun;Lee, Chang-Joo
    • Journal of Aquaculture
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    • v.18 no.2
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    • pp.81-85
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    • 2005
  • We isolated two parvalbumin cDNAS by expressed sequence tag analysis (1,577 ESTs in total) from the self-fertilizing fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae). Two isoforms of parvalbumin genes showed high similarity to those of carp at 88% and 91% amino acid residues identity, respectively, and showed 79.8% similarity between two parvalbumin isoforms. Of 1,577 ESTs from R. marmroatus sequenced, parvalbumin 1 gene was most abundant. This gene was strongly expressed in the order of muscle, eye, and brain, while it was expressed slightly in other tissues. In this paper, we discussed on the R. marmoratus parvalbumin genes on its sequence and basic characteristics.

Involvement of $PLA_2$ Isoforms in Muscarinic Receptor-Mediated sAPP Release and Store-Operated Calcium Entry in SH-SY5Y Cells.

  • Cho, Hye-Won;Kim, Jin-Hyung;Choi, Shin-Kyu;Kim, Hwa-Jung
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.128.2-129
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    • 2003
  • We previously reported that phospholipase$A_2$ ($PLA_2$)-related pathway and capacitative calcium entry (CCE) via store-operated calcium channel (SOC) were involved in the regulation of muscarinic receptor- mediated sAPP release. We also observed that stimulation of muscarinic receptor associated with the inositol phosphate cascade resulted not only in increase of CCE but also in activation of PLA$_2$ in SH-SY5Y cells. In this study, we further investigated whether the $PLA_2$ isoforms differently regulate the muscarinic receptor-mediated sAPP release, and examined the relationships between activation of $PLA_2$ isoforms and CCE mediated by muscarinic receptors in SH-SY5Y cells. (omitted)

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Novel splice isoforms of pig myoneurin and their diverse mRNA expression patterns

  • Guo, Xiaohong;Li, Meng;Gao, Pengfei;Cao, Guoqing;Cheng, Zhimin;Zhang, Wanfeng;Liu, Jianfeng;Liu, Xiaojun;Li, Bugao
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.10
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    • pp.1581-1590
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    • 2018
  • Objective: The aim of this study was to clone alternative splicing isoforms of pig myoneurin (MYNN), predict the structure and function of coding protein, and study temporal and spatial expression characteristics of each transcript. Methods: Alternative splice isoforms of MYNN were identified using RNA sequencing (RNA-seq) and cloning techniques. Quantitative real-time polymerase chain reaction (qPCR) was employed to detect expression patterns in 11 tissues of Large White (LW) and Mashen (MS) pigs, and to study developmental expression patterns in cerebellum (CE), stomach (ST), and longissimus dorsi (LD). Results: The results showed that MYNN had two alternatively spliced isoforms, MYNN-1 (GenBank accession number: KY470829) and MYNN-2 (GenBank accession number: KY670835). MYNN-1 coding sequence (CDS) is composed of 1,830 bp encoding 609 AA, whereas MYNN-2 CDS is composed of 1,746 bp encoding 581 AA. MYNN-2 was 84 bp less than MYNN-1 and lacked the sixth exon. MYNN-2 was found to have one $C_2H_2$ type zinc finger protein domain less than MYNN-1. Two variants were ubiquitously expressed in all pig tissues, and there were significant differences in expression of different tissues (p<0.05; p<0.01). The expression of MYNN-1 was significantly higher than that of MYNN-2 in almost tissues (p<0.05; p<0.01), which testified that MYNN-1 is the main variant. The expression of two isoforms decreased gradually with increase of age in ST and CE of MS pig, whereas increased gradually in LW pig. In LD, the expression of two isoforms increased first and then decreased with increase of age in MS pig, and decreased gradually in LW pig. Conclusion: Two transcripts of pig MYNN were successfully cloned and MYNN-1 was main variant. MYNN was highly expressed in ST, CE, and LD, and their expression was regular. We speculated that MYNN plays important roles in digestion/absorption and skeletal muscle growth, whereas the specific mechanisms require further elucidation.

Characterization of Monoclonal Antibodies against Human Leukocyte Common Antigen (CD45)

  • Shin, Hyang-Mi;Cho, Woon-Dong;Lee, Geon-Kook;Lee, Seon-Hwa;Lee, Kyung-Mee;Ji, Gil-Yong;Yoon, Sang-Soon;Koo, Ji-Hae;Lee, Ho-Chang;Lee, Ki-Hyeong;Song, Hyung-Geun
    • IMMUNE NETWORK
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    • v.11 no.2
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    • pp.114-122
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    • 2011
  • Background: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. Methods: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. Results: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). Conclusion: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen.

Human Vesicular Glutamate Transporters Functionally Complement EAT-4 in C. elegans

  • Lee, Dukgyu;Jung, Sunki;Ryu, Jungmin;Ahnn, Joohong;Ha, Ilho
    • Molecules and Cells
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    • v.25 no.1
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    • pp.50-54
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    • 2008
  • The vesicular glutamate transporter (VGLUT) transports glutamate into pre-synaptic vesicles. Three isoforms of VGLUT have been identified in humans, but their functional differences remain largely unknown. EAT-4 is the only homologue of human VGLUT in C. elegans. Here we report that mutants of eat-4 exhibit hyperforaging behavior and that each of the isoforms of human VGLUT functionally rescues the defects in eat-4 worms.

Changes in Expression of Connexin Isoforms in the Caudal Epididymis of Adult Sprague-Dawley Rats exposed to Estradiol Benzoate or Flutamide at the Neonatal Age

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.20 no.3
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    • pp.237-245
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    • 2016
  • Direct communication between neighboring cells via gap junction in tissue is important for maintenance and regulation of its physiological functions. Each epididymal region has different composition of cell types. It is well recognized that the epididymis is a steroid hormone-responsive tissue. The present study was designed to determine the effect of estradiol benzoate (EB) or flutamide exposured at the early postnatal age on the expression of connexin (Cx) isoforms in the caudal epididymis. The EB or flutamide was subcutaneously administrated to male Spragure Dawley rat at 7 days of age, and expressional changes of Cx isoforms in the adult corpus epididymis were determined by quantitative real-time PCR. The treatment of low-dose EB resulted in decreases of Cx30.3, Cx31.1, Cx37, and Cx45 expression but caused an increase of Cx32 expression. Exposure to high-dose EB led into expressional increases of Cx31, Cx31.1, Cx32, Cx40, and Cx43, even though a decrease of Cx37 expression was found with a high-dose EB treatment. A low-dose flutamide induced increases of Cx31, Cx31.1, Cx32, and Cx43 expression but a decrease of Cx37 expression. Expression of most Cx genes were significantly increased by a high-dose flutamide, while no expressional change of Cx26 and Cx40 was detected by a high-dose flutamide. These results indicate that expression of Cx isoforms in the caudal epididymis is altered by exposure to steroidal compounds at the prepubertal age. It is suggested that a contact with environmental exogenous materials during the early postnatal period would lead to alteration of epididymal functions at the adult.

Exogenous Exposure to Estradiol Benzoate or Flutamide at the Weaning Age Alters Expression of Connexin Isoforms in the Initial Segment of Male Rat

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.19 no.1
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    • pp.43-51
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    • 2015
  • Connexin (Cx) is a complex which allows direct communication between neighboring cells via exchange of signaling molecules and eventually leads to functional harmony of cells in a tissue. The initial segment (IS) is an excurrent duct of male reproductive tract and expression of numerous genes in the IS are controlled by androgens and estrogens. The effects of these steroid hormones on gene expression in the IS during postnatal development have not extensively examined. The present research investigated expressional modulation of Cx isoforms in the IS by exogenous exposure to estrogen agonist, estradiol benzoate (EB), or androgen antagonist, flutamide (Flu), at weaning age. Two different doses of EB or Flu were subcutaneously administrated in 21-day old of male rats, and expressional changes of Cx isoforms in the adult IS were analyzed by quantitative real-time PCR. Treatment of a low-dose EB ($0.015{\mu}g/kg$ body weight) resulted in an increased expression of Cx31 gene and a decreased expression of Cx37 gene. A high-dose EB ($1.5{\mu}g/kg$ body weight) treatment caused an increase of Cx31 gene expression. Increased levels of Cx30.3 and Cx40 transcripts were observed with a low-dose Flu ($500{\mu}g/kg$ body weight) treatment. Treatment of high-dose Flu (50 mg/kg body weight) led to expressional increases of Cx30.3, 40, and 43 genes. Our previous and present findings suggest differential responsiveness on gene expression of Cx isoforms in the IS by androgens and estrogens at different postnatal ages.