• Parasites, Hosts and Diseases
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• v.46 no.4
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• pp.209-216
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• 2008

A monoclonal antibody against Toxoplasma gondii of Tg556 clone (Tg556) blotted a 29 kDa protein, which was localized in the dense granules of tachyzoites and secreted into the parasitophorous vacuolar membrane (PVM) after infection to host cells. A cDNA fragment encoding the protein was obtained by screening a T. gondii cDNA expression library with Tg556, and the full-length was completed by 5'-RACE of 2,086 bp containing an open reading frame (ORF) of 669 bp. The ORF encoded a polypeptide of 222 amino acids homologous to the revised GRA3 but not to the first reported one. The polypeptide has 3 hydrophobic moieties of an N-terminal stop transfer sequence and 2 transmembrane domains (TMD) in posterior half of the sequence, a cytoplasmic localization motif after the second TMD and an endoplasmic reticulum (ER) retrival motif in the C-terminal end, which suggests GRA3 as a type III transmembrane protein. With the ORF of GRA3, yeast two-hybrid assay was performed in HeLa cDNA expression library, which resulted in the interaction of GRA3 with calcium modulating ligand (CAMLG), a type II transmembrane protein of ER. The specific binding of GRA3 and CAMLG was confirmed by glutathione S-transferase (GST) pull-down and immunoprecipitation assays. The localities of fluorescence transfectionally expressed from GRA3 and CAMLG plasmids were overlapped completely in HeLa cell cytoplasm. In immunofluorescence assay, GRA3 and CAMLG were shown to be co-localized in the PVM of host cells. Structural binding of PVM-inserted GRA3 to CAMLG of ER suggested the receptor-ligand of ER recruitment to PVM during the parasitism of T. gondii.

• Journal of Plant Biotechnology
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• v.3 no.1
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• pp.27-31
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• 2001

New types of cytoplasmic male sterility in Brassica species would be very useful for the production of F$_1$, hybrid seeds. Leaves and stems of rapid cycling stock of B.juncea (CrGC4-3) containing Anand CMS were used as experimental materials for plant regeneration from protoplast culture. Very high plant regeneration rate (85%) was found in the Kao & Michayluk medium supplemented with 2 mg/L zeatin, 0.5 mg/L BAP, and 1 mg/L NAA when only leaf, not stem, segments were cultured. Protoplasts were isolated from leaves using mixtures of enzymes (1% Cellulycin, 0.5% Macerozyme) in 0.4 M mannitol and 50 mM $CaCl_2$.$2H_2$O. Mcrocalli induced from protoplasts were transferred to the shoot regeneration medium containing 2 mg/L BAP, 2 mg/L zeatin, and 0.5 mg/L NAA. After 60 days of initial protoplast culture, regenerated plantlets were obtained, acclimatized, transplanted into the pots, and grown up to the flowering stage.

• Journal of Plant Biology
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• v.38 no.2
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• pp.143-151
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• 1995

Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

• Journal of Crop Science and Biotechnology
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• v.10 no.2
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• pp.86-91
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• 2007

A representative group of 38 improved basmati lines including maintainers of sterile lines were studied for genetic diversity utilizing Mahalanobis $D^2$ statistics. A wide diversity was observed having ten clusters with high intra- and inter-cluster distance. Heterosis was estimated utilizing the cytoplasmic male sterile lines from the clusters having high intra- and inter-cluster distance. Highly heterotic hybrids were obtained from the hybridization programme. Cross combinations IR68281A/Pusa 1235-95-73-1-1, IR68281A/RP 3644-41-9-5, Pusa 3A/UPR 2268-4-1, IR 68281A/Pusa Basmati-1, IR68281A/BTCE 10-98, and IR58025A/HKR 97-401 were found to be highly heterotic for grain yield/plant with other agronomic and quality traits. Additionally, a positive association of intra-cluster distance with heterosis was observed, which could be utilized as a guideline for predicting heterosis in basmati hybrid rice breeding program. Also, a positive association between inter-cluster distance and heterosis was observed.

• BMB Reports
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• v.43 no.6
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• pp.432-437
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• 2010

LBH is a transcription factor as a candidate gene for CHD associated with partial trisomy 2p syndrome. To identify potential LBH-interacting partners, a yeast two-hybrid screen using LBH as a bait was performed with a human heart cDNA library. One of the clones identified encodes ${\alpha}B$-crystallin. Co-immunoprecipitation and GST pull-down assays showed that LBH interacts with ${\alpha}B$-crystallin, which is further confirmed by mammalian two-hybrid assays. Co-localization analysis showed that in COS-7 cells, ${\alpha}B$-crystallin that is cytoplasmic alone, accumulates partialy in the nucleus when co-transfected with LBH. Transient transfection assays indicated that overexpression of LBH or ${\alpha}B$-crystallin reduced the transcriptional activities of p53 and p21, respectively, Overexpression of both ${\alpha}B$-crystallin and LBH together resulted in a stronger repression of the transcriptional activities of p21 and p53. These results showed that the interaction of LBH and ${\alpha}B$-crystallin may inhibit synergistically the transcriptional regulation of p53 and p21.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.4
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• pp.420-427
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• 1995

This study was carried out to investigate heterosis in photosynthetic characteristics and yield of F$_1$ hybrid rice using genetic cytoplasmic male sterile lines. Two F$_1$ hybrid rices, HR7028A/Yongmoonbyeo and IR54756A/Yongmoonbyeo, showed heterobeltiosis in dry weight and leaf area at the heading date and at the 20 day after heading. HR7028A/Yongmoonbyeo showed heterobeltiosis and IR54756A/Yongmoonbyeo showed heterosis in chlorophyll content and RuDPCase activity in flag leaf, T-sugar and starch content in stem, and $CO_2$ fixation at the 20 day after heading as well as at the heading date. HR7028A/Yongmoonbyeo showed heterobeltiosis in number of panicles and grains per panicle, and IR54756A/Yongmoonbyeo had heterobeltiosis in number of panicles per plant. As results, heterobeltiosis of HR7028A/Yongmoonbyeo in yield was 11% and that of IR54756A/Yongmoonbyeo was 4%.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.30 no.4
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• pp.427-430
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• 1985

Extrusion degree of stigma, numbers of opened florets at different time during anthesis, angle of opening florets and outcrossed seed set percent were investigated in four male sterile rices having background of Korean cultivars; Suwon 296A, Suwon 304A, Suwon 310A and Suwon 296GMS, and the Chinese cytoplasmic genetic male sterile rice Zhenshan 97A as control in order to get basic informations for hybrid rice breeding. The results obtained were summarized as follows; Extrusion degree of stigma in the four Korean male sterile lines tested was lower than in the Chinese one Zhenshan 97A. The better extrusion of stigma showed the higher outcross rate in male sterile rices. There were almost no relationships between peak of anthesis and outcrossed seed set in the male sterile rices tested. Large anthesis angle with long anthesis duration appeared to be essential for high outcrossed seed set of the male sterile rices. The genetic male sterile line showed the larger anthesis angle than the cytoplasmic and genetic male sterile one in the lines having the same background to the Korean rice cultivar Suwon 296.

• Journal of Korean Society of Forest Science
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• v.46 no.1
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• pp.53-56
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• 1980

We observed performance of segregation in flower color and type of hybrids which obtained from crossing of intra species in Hibiscus syriacus. Obtained results were followings. 1. The purple flower was dominance to the white one and this was presumed that was owing to cytoplasmic heredity. 2. Single and double petal of flower was presumed that was originated from factors of Ss and dd. 3. There was not variation in flower color and type of $F_1$ hybrid between 4 n and 2 n Hibiscus syriacus. 4. There were many variation of flower color among $F_1$ hybrids which abtained in open pollination of Hibiscus syriacus. 5. We could observe many flowers variegated with red color among $F_1$ hybrids which obtained in crossing between double petal flowers of Hibiscus syriacus.

• Horticultural Science & Technology
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• v.33 no.4
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• pp.550-558
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• 2015

To analyze the linkage relationships among molecular markers recently reported to be linked to onion (Allium cepa L.) Ms, a restorer-of-fertility locus, in onion (Allium cepa L.), three single nucleotide polymorphism markers were converted into cleaved amplified polymorphic sequence (CAPS) markers based on onion transcriptome sequences and the rice genome database. Analysis of the recombinants selected from 4,273 segregating plants using CAPS and other linked markers demonstrated the jnurf13 and jnurf610 markers to perfectly co-segregate with the Ms locus. In contrast to jnurf13, the jnurf610 marker was not in perfect linkage disequilibrium with the Ms locus in diverse breeding lines. Thus, the jnurf13 marker and the marker for identification of cytoplasm types were utilized to enhance the efficiency of onion breeding through four applications. First, 89 maintainer lines containing the normal cytoplasm and homozygous recessive Ms genotypes were successfully identified from 100 breeding lines. Second, these two molecular markers were used to analyze the main sources of male-fertile contaminants frequently found in the male-sterile parental lines during F1 hybrid seed production. The majority of the contaminants contained heterozygous Ms genotypes, indicating that pollen grains harboring the dominant Ms genotype may have been introduced during propagation of the maintainer lines. Therefore, the genetic purity of the two maintainer lines was analyzed in the third application, and the results showed that both maintainer lines contained 13-21% off-types. Finally, the two markers were used to increase the seed yield potentials of two open-pollinated varieties containing sterile cytoplasms by removing the plants harboring homozygous recessive and heterozygous Ms genotypes.

• BMB Reports
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• v.52 no.7
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• pp.445-450
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• 2019

TTYH2 is a calcium-activated, inwardly rectifying anion channel that has been shown to be related to renal cancer and colon cancer. Based on the topological prediction, TTYH2 protein has five transmembrane domains with the extracellular N-terminus and the cytoplasmic C-terminus. In the present study, we identified a vesicle transport protein, ${\beta}$-COP, as a novel specific binding partner of TTYH2 by yeast two-hybrid screening using a human brain cDNA library with the C-terminal region of TTYH2 (TTYH2-C) as a bait. Using in vitro and in vivo binding assays, we confirmed the protein-protein interactions between TTYH2 and ${\beta}$-COP. We also found that the surface expression and activity of TTYH2 were decreased by co-expression with ${\beta}$-COP in the heterologous expression system. In addition, ${\beta}$-COP associated with TTYH2 in a native condition at a human colon cancer cell line, LoVo cells. The over-expression of ${\beta}$-COP in the LoVo cells led to a dramatic decrease in the surface expression and activity of endogenous TTYH2. Collectively, these data suggested that ${\beta}$-COP plays a critical role in the trafficking of the TTYH2 channel to the plasma membrane.