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41.
Establishment of a cell line with high transfection efficiency from zebrafish Danio rerio embryos and its susceptibility to fish viruses
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Y. L. Jin L. M. Chen Y. Le Y. L. Li Y. H. Hong K. T. Jia M. S. Yi 《Journal of fish biology》2017,91(4):1018-1031
A cell line ZBE3 isolated from a continuous cell culture derived from zebrafish Danio rerio blastomeres by clonal growth was characterized. ZBE3 cells had been subcultured for >120 passages since the initial primary culture of the blastomeres. The ZBE3 cells grow stably at temperature from 20 to 32° C with an optimum temperature of 28° C in ESM2 or ESM4 medium with 15% foetal bovine serum (FBS). The optimum FBS concentration for ZBE3 cell growth ranged from 15 to 20%. Cytogenetical analysis indicated that the modal chromosome number of ZBE3 cells was 50, the same as the diploid chromosome number of D. rerio. Significant cytopathic effect was observed in ZBE3 cells after infection with redspotted grouper nervous necrosis virus, Singapore grouper iridovirus and grass carp reovirus, and the viral replication in the cells was confirmed by real‐time quantitative PCR and transmission electron microscopy, indicating the susceptibility of ZBE3 cells to the three fish viruses. After transfected with pEGFP‐N3 plasmid, ZBE3 cells showed a transfection efficiency of about 40% which was indicated by the percentage of cells expressing green fluorescence protein. The stable growth, susceptibility to fish viruses as well as high transfection efficiency make ZBE3 cells be a useful tool in transgenic manipulation, fish virus‐host cell interaction and immune response in fish. 相似文献
42.
Hui Ge Kebing Lin Mi Shen Shuiqing Wu Yilei Wang Ziping Zhang Zhiyong Wang Yong Zhang Zhen Huang Chen Zhou Qi Lin Jianshao Wu Lei Liu Jiang Hu Zhongchi Huang Leyun Zheng 《Molecular ecology resources》2019,19(6):1461-1469
The red‐spotted grouper Epinephelus akaara (E. akaara) is one of the most economically important marine fish in China, Japan and South‐East Asia and is a threatened species. The species is also considered a good model for studies of sex inversion, development, genetic diversity and immunity. Despite its importance, molecular resources for E. akaara remain limited and no reference genome has been published to date. In this study, we constructed a chromosome‐level reference genome of E. akaara by taking advantage of long‐read single‐molecule sequencing and de novo assembly by Oxford Nanopore Technology (ONT) and Hi‐C. A red‐spotted grouper genome of 1.135 Gb was assembled from a total of 106.29 Gb polished Nanopore sequence (GridION, ONT), equivalent to 96‐fold genome coverage. The assembled genome represents 96.8% completeness (BUSCO) with a contig N50 length of 5.25 Mb and a longest contig of 25.75 Mb. The contigs were clustered and ordered onto 24 pseudochromosomes covering approximately 95.55% of the genome assembly with Hi‐C data, with a scaffold N50 length of 46.03 Mb. The genome contained 43.02% repeat sequences and 5,480 noncoding RNAs. Furthermore, combined with several RNA‐seq data sets, 23,808 (99.5%) genes were functionally annotated from a total of 23,923 predicted protein‐coding sequences. The high‐quality chromosome‐level reference genome of E. akaara was assembled for the first time and will be a valuable resource for molecular breeding and functional genomics studies of red‐spotted grouper in the future. 相似文献
43.
Tianyun Shao Xiaoyan Chen Dingxuan Zhai Tao Wang Xiaohua Long Zhaopu Liu 《Journal of fish biology》2019,95(4):1022-1029
This study was aimed at evaluating the physiological and metabolic responses of juvenile hybrid grouper ♀ Epinephelus fuscoguttatus × ♂ Epinephelus lanceolatus to stocking density. Hybrid grouper juveniles (mean ± SE = 25.43 ± 2.36 g live mass) were stocked for 22 weeks in a recirculating aquaculture system (RAS) under four different densities: low stocking density (LD; 1.03 kg m−3), medium stocking density (MD; 2.06 kg m−3), high stocking density (HD; 3.09 kg m−3) and extra-high stocking density (EHD; 4.11 kg m−3). Biometric variables were recorded and plasma, liver, intestine and stomach samples were taken for biochemical analysis at the end of the experimental period. Final stocking densities were 6.27, 16.04, 23.77 and 28.32 kg m−3, respectively, with significant differences in growth performance. Our results showed that the best growth rates and feed utilisation occurred in the MD group. Higher plasma cortisol and glucose levels and lower triglyceride levels reflected the stress responses in the EHD group. Moreover, the activity of aspartate and alanine transaminases was elevated in the HD and EHD groups due to enhanced gluconeogenesis. The activity of the digestive enzyme pepsin significantly increased in the MD group. We found that 2.06–3.09 kg m−3 is the most suitable starting density for culturing juvenile hybrid grouper in recirculating aquaculture systems. 相似文献
44.
Occurrence of dusky grouper Epinephelus marginatus (Lowe, 1834) in gulfs of northern Patagonia, Argentina 总被引:1,自引:0,他引:1
The occurrence of the dusky grouper Epinephelus marginatus in northern Patagonian Gulfs San Matías and Nuevo, Argentina, is recorded. This finding represents the southernmost record of the species for the southern hemisphere. 相似文献
45.
Combination of microwell structures and direct oxygenation enables efficient and size‐regulated aggregate formation of an insulin‐secreting pancreatic β‐cell line
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Marie Shinohara Hiroshi Kimura Kevin Montagne Kikuo Komori Teruo Fujii Yasuyuki Sakai 《Biotechnology progress》2014,30(1):178-187
Spherical three‐dimensional (3D) cellular aggregates are valuable for various applications such as regenerative medicine or cell‐based assays due to their stable and high functionality. However, previous methods to form aggregates have shown drawbacks, being labor‐intensive, showing low productivity per unit area or volume and difficulty to form homogeneous aggregates. We proposed a novel strategy based on oxygen‐permeable polydimethylsiloxane (PDMS) honeycomb microwell sheets, which can theoretically supply about 80 times as much oxygen as conventional polystyrene culture dishes, to produce recoverable aggregates in controllable sizes using mouse insulinoma cells (MIN6‐m9). In 48 hours of culture, the PDMS sheets produced aggregates whose diameters were strictly controlled (?32, 60, 90, 150 and 280 mm) even at an inoculum density eight times higher (8.0×105 cells/cm2) than that of normal confluent monolayers (1.0×105 cells/cm2). Measurement of the oxygen tension near the cell layer and glucose/lactate analysis clearly showed that cells exhibit aerobic respiration on the PDMS‐based culture system. Glucose‐responsive insulin secretion of the recovered aggregates showed that the aggregates around 90 mm in diameter secreted the largest amounts of insulin. This confirmed the advantages of 3D cellular organization and the existence of a suitable aggregate size, above which excess organization leads to a decreased metabolic response. These results demonstrated that this microwell‐based PDMS culture system provides a promising method to form size‐regulated and better functioning 3D cellular aggregates of various kinds of cells with a high yield per surface area. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:178–187, 2014 相似文献
46.
We investigated an integrated manufacturing method to develop lightweight composite materials. To design the preparation process for the integrated honeycomb plates, the shear and compressive mechanical properties of the corresponding composite materials were also investigated. The results indicate that these composite materials, with two types of resins reinforced by short basalt fibers, exhibit obvious toughness, particularly in their compressive properties. The Epoxy Resin (ER) is denser and has better bonding at the fiber and matrix interface than the vinyl ester resin matrix. Therefore, the ER exhibits better shear and compressive strengths than the vinyl ester resin matrix, thereby providing a design technology of the preparation process of the integrated honeycomb plate. The matrix material of this plate is composed of an epoxy (E51 ), a curing agent (593), and a thinner (501) at a ratio of 10:3:1; short basalt fibers are added as a reinforcing material at a 30% volume fraction. By increasing the curing temperature and other experimental conditions, we obtained an expected integrated honeycomb plate. This integrated honeycomb plate possesses properties such as a high fiber content, good shear and compressive performance, and high proc- essing efficiency. 相似文献
47.
Lili Zhao Changwei Shao Xiaolin Liao Hongyu Ma Xuechun Zhu Songlin Chen 《Conservation Genetics》2009,10(3):743-745
Yellow grouper (Epinephelus awoara) is a commercially important marine fish species. A dinucleotide-enriched genomic library of E. awoara was constructed using the method of FIASCO. Twelve loci were polymorphic in a test population with alleles per locus ranging
from two to eight, and observed and expected heterozygosities per locus from 0.13 to 1.00 and from 0.20 to 0.86, respectively.
Five loci significantly deviated from Hardy–Weinberg equilibrium and no significant linkage disequilibrium was found between
pairs of loci. Cross-species amplification of these polymorphic microsatellite loci was performed in additional two related
species. These polymorphic microsatellite loci would be useful for investigating genetic diversity of E. awoara and related species.
L. Zhao and C. Shao have contributed equally to this work. 相似文献
48.
Seven-band grouper (Epinephelus septemfasciatus) is a commercially important fishery species. Sixty-six microsatellite loci were isolated from a dinucleotide-enriched genomic
library of E. septemfasciatus. Twelve of these loci were polymorphic in a test population with alleles per locus ranging from two to five, and observed
and expected heterozygosities per locus from 0.04 to 1.00 and from 0.28 to 0.76, respectively. Three loci significantly deviated
from Hardy–Weinberg equilibrium and no significant linkage disequilibrium was found between pairs of loci after Bonferroni
correction. Cross-species amplification of these polymorphic microsatellite loci was performed in additional two related species.
These polymorphic microsatellite loci would be useful for investigating genetic resource of E. septemfasciatus and other related species.
Lili Zhao and Changwei Shao have contributed equally. 相似文献
49.
Colloidal lithography was used to make a novel array (2-D) of micro-rings, dots, and interconnected-honeycomb structures.
These geometries are controlled using the curing temperature-dependent rheological properties of the siloxane elastomer precursor.
Serratia marcescens was patterned on the interconnected honeycomb microstructure demonstrating a potential application for microbioanalytical
devices, microfluidics, and bio-micro-electromechanical systems.
Received 26 August 2005; Revisions requested 23 September 2005; Revisions received 10 November 2005; Accepted 11 November
2005 相似文献
50.
Alam MA Bhandari RK Kobayashi Y Nakamura S Soyano K Nakamura M 《Molecular reproduction and development》2006,73(2):206-214
11-ketotestosterone (11-KT), a potent male-specific androgen in fish, has important roles on spermatogenesis, male behavior, and nuptial coloration. The site of 11-KT synthesis and its role on male germ cell development during protogynous sex change is not clearly understood. We examined the dynamics of steroidogenic enzymes immunolocalization, viz cholesterol side-chain cleavage (P450scc), biomarker of steroids and cytochrome P45011beta-hydroxylase (P45011beta), downstream to 11-KT production, throughout the process of sex change in honeycomb grouper (Epinephelus merra). In female, P450scc immunoreactivity (-ir) was observed in the theca layer and tunica near blood vessels (BV). During the onset of sex change, P450scc reactive cells were observed in the remaining follicle layer of degenerated oocyte of the ovo-testis in early transitional (ET) and late transitional (LT). In male, P450scc-ir was localized in the interstitial Legdig cells of testis. P45011beta reactive cells were observed in the tunica near BV in female but not in theca layer. In ET and LT phases gonads, P45011beta localized in remaining follicle layer of degenerated oocyte and tunica near BV. On the other hand, in male, both interstices and tunica near BV showed strong signals against P45011beta. Moreover, in vivo and in vitro levels of 11-KT related with the changes in the nuclei diameter of P45011beta-positive cells in both tunica near BV and remaining follicle layer of degenerated oocyte to interstices during the progress of sex change. The present results suggest that 11-KT produced in the tunica near BV may provide the stimulus for female to degenerate oocytes and initiate sex change. However, 11-KT produced both in tunica near BV and remaining follicle layer of degenerated oocyte possibly plays critical role during testicular differentiation as well as gonadal restructuring at mid to late phases (ET to LT) of sex change in honeycomb grouper. 相似文献