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1.
To reveal the key factor in self-healing from LCDV (lymphocystis disease virus)-infec Japanese flounder (Paralichthys olivaceus), serum proteins from self-healing and sick Japanese floun were separated by two-dimensional electrophoresis to screen differentially expressed proteins. Prot spots demonstrating changes greater than two-fold in the expression level were digested and furt identified in capillary liquid chromatography tandem mass spectrometry (LC-MS/MS). Two immuni relevant proteins were thus identified as transferrin and the complement component C3 of Japan flounder. These findings suggest that the two proteins may play important roles in the self-healing lymphocystis in Japanese flounder. This is an important theoretical foundation to promote self-healing LCDV-infected Japanese flounder by improving their innate immunity. 相似文献
2.
The molecular basis of color polymorphism in the shells of the pearl oyster Pinctada fucata is largely unknown. We developed a red-shelled family line and used suppression subtractive hybridization (SSH) to screen for differentially expressed genes in red- and non-red-shelled pearl oysters. We constructed forward and reverse cDNA subtractive libraries consisting of 2 506 and 797 clones, respectively. Among 343 randomly selected clones in the forward library, 304 sequences were identified in GenBank using BLASTx and BLASTn. Of the 304 sequences, 13 showed no similarity to known sequences and 291 were matched with known genes of the pearl oyster, including shematrin-1, shematrin-2, shematrin-6, shematrin-7, nacrein, nacrein-like protein, aspein for shell matrix protein, glycine-rich protein, mantle gene 5, 28S, EST00031, EST00036, 16S, and COΙ. In the reverse library, 7 clones were sequenced and analyzed by BLAST. Two sequences shared similarity with EST00036 from the P. fucata subtraction cDNA library, four with the P. fucata mitochondrial gene for 16S rRNA and 1 with P. fucata shematrin-2. We evaluated the expression of 12 genes from the forward library using RT PCR. Two sequences matched with 16S and COΙ so were considered to be false positives. The remaining 10 sequences were differentially expression in the red-shelled pearl oysters. Our results suggest that differential expression of these genes may be related to color variation in the red-shelled family line of the pearl oyster. 相似文献
3.
Xianhong Meng Xiaoli Shi Jie Kong Sheng Luan Kun Luo Baoxiang Cao Ning Liu Xia Lu Xupeng Li Kangyu Deng Jiawang Cao Yingxue Zhang Hengheng Zhang 《中国海洋大学学报(英文版)》2017,16(5):863-872
To elucidate the molecular response of shrimp hepatopancreas to white spot syndrome virus (WSSV) infection, microarray was applied to investigate the differentially expressed genes in the hepatopancreas of ‘Huanghai No. 2’ (Fenneropenaeus chinensis). A total of 59137 unigenes were designed onto a custom-made 60K Agilent chip. After infection, the gene expression profiles in the hepatopancreas of the shrimp with a lower viral load at early (48–96 h), peak (168–192 h) and late (264–288 h) infection phases were analyzed. Of 18704 differentially expressed genes, 6412 were annotated. In total, 5453 differentially expressed genes (1916 annotated) expressed at all three phases, and most of the annotated were either up- or down-regulated continuously. These genes function diversely in, for example, immune response, cytoskeletal system, signal transduction, stress resistance, protein synthesis and processing, metabolism among others. Some of the immune-related genes, including antilipopolysaccharide factor, Kazal-type proteinase inhibitor, C-type lectin and serine protease encoding genes, were up-regulated after WSSV infection. These genes have been reported to be involved in the anti-WSSV responses. The expression of genes related to the cytoskeletal system, including β-actin and myosin but without tubulin genes, were down-regulated after WSSV infection. Astakine was found for the first time in the WSSV-infected F. chinensis. To further confirm the expression of differentially expressed genes, quantitative real-time PCR was performed to test the expression of eight randomly selected genes and verified the reliability and accuracy of the microarray expression analysis. The data will provide valuable information to understanding the immune mechanism of shrimp’s response to WSSV. 相似文献
4.
Members of the DnaJ family are proteins that play a pivotal role in various cellular processes, such as protein folding, protein transport and cellular responses to stress. In the present study, we identified and characterized the full-length DnaJ cDNA sequence from expressed sequence tags of Pyropia yezoensis (PyDnaJ) via rapid identification of cDNA ends. This cDNA encoded a protein of 429 amino acids, which shared high sequence similarity with other identified DnaJ proteins, such as a heat shock protein 40/DnaJ from Pyropia haitanensis. The relative mRNA expression level of PyDnaJ was investigated using real-time PCR to determine its specific expression during the algal life cycle and during desiccation. The relative mRNA expression level in sporophytes was higher than that in gametophytes and significantly increased during the whole desiccation process. These results indicate that PyDnaJ is an authentic member of the DnaJ family in plants and red algae and might play a pivotal role in mitigating damage to P. yezoensis during desiccation. 相似文献
5.
光温敏核不育水稻幼穗育性转换期蛋白质组的初步分析 总被引：2，自引：0，他引：2
为找寻光温敏核不育水稻幼穗与育性调控有关的蛋白质,采用固相pH梯度-SDS聚丙烯酰胺双向凝胶电泳对光温敏核不育水稻(培矮64S)育性转换期幼穗总蛋白进行了分离,获取凝胶图像并进行重复性分析,通过微量测序-Edman降解方法和原位酶解及MALDI-TOF质谱的方法鉴定了60个蛋白质点。结果表明:本方法可得取分辨率和重复性较好的凝胶图谱。蛋白质点在2D胶上的重复性为:沿等电聚焦方向偏差为1.49±0.22 mm,沿SDS-PAGE方向偏差为1.24±0.18 mm。37个蛋白质点得到了准确的鉴定结果,其中16个是差异蛋白质点。在不育变化为可育的过程中,明显表达上调的蛋白质点包括几丁质酶,丙酮酸脱氢酶等;明显下调的蛋白质包括超氧化物歧化酶,硝酸还原酶脱辅基酶蛋白等;金属硫蛋白RicMT只在可育中存在。 相似文献
6.
In crustaceans, the male sexual differentiation and maintenance are specially regulated by androgenic gland (AG). However, little is known about the genes involved in the regulation process. RNA-Seq was performed on AG with ejaculatory duct (AG_ED) and ejaculatory duct (ED) as control in Eriocheir sinensis, one of the most important economic and fishery crabs with typically sex dimorphism. A total of 925 unigenes were identified as differentially expressed genes (DEGs) and the expression of nine genes randomly selected was confirmed by qRT-PCR. 667 unigenes were up-regulated in AG_ED, being supposed to be AG preferential genes. Among them, the full length of insulin-like androgenic gland factor (IAG) cDNA named as Es-IAG was obtained as a logo gene of AG, which together with the genes insulin-like receptor (INR), and single insulin binding domain protein (SIBD), might constitute the sex regulation pathway. Several sex related genes were identified, and their function will have to be investigated. Also, the identification of juvenile hormone epoxide hydrolase 1 (JHEH1), ecdysteroid 22-hydroxylase (DIB) and ecdysone receptor (ECR) preliminarily clarified the molecular regulation mechanism of eyestalk-AG-testis axis, which plays important roles in molting and reproduction. The results will enhance our understanding for the molecular basis of the AG involved in male sex regulation in crabs.
相似文献7.
The carbohydrate and lipid contents in two planktonic crustaceans, i. e.Pseudeuphausia sinica andAcetes chinensis sampled from Xiamen coastal waters were estimated. InP. sinica, the contents of carbohydrates (in dry weight) and lipids varied from 2.19–2.33% and 21.2–21.9% respectively; inA. chinensis from 1.74–2.55% and 14.43–15.10% respectively. The analyses of fatty acids by gas-chromatograph (Model 103) showed that eight
fatty acids of 14∶0, 15∶0, 16∶0, 17∶0, 18∶0, 18∶2, 20∶2 and 22∶2 were found in these two animals, that 16∶0, 18∶0, 18∶2, and
20∶2 formed the major constituents with 91.08% (P. sinica) and 74.80% (A. chinensis) of the total fatty acids, and that the values of odd carbon types (15∶0 and 17∶0) were fairly low, 4.47% and 3.36% respectively.
Three monosaccharides: glucose, fructose, and galactose detected by high-liquid chromatograph (Waters 208) accounted for more
than 60% of the total carbohydrates, especially the glucose, exceeding by 40%. 相似文献
8.
Shuo Li Chunmei Li Xubo Wang Yanan Wang Zhipeng Liu Teng Zhai Quanqi Zhang 《中国海洋大学学报(英文版)》2010,9(1):59-64
A novel immune-related gene was expressed in Japanese flounder (Paralichthys olivaceus) injected with Vibrio anguillarum. The complete cDNA contained a 169 bp 5’UTR, a 336 bp open reading frame (ORF) encoding 111 amino acids and a 556bp 3’UTR.
Six exons and five introns were identified in the PoIR2 gene. Blastp similarity comparison showed its encoding protein had 50% similarity to Danio rerio neuromedin S (NMS), but further alignment indicated they did not have NMS C-terminal conservational signature domain. So
it was not defined as an NMS homologue. Protein structure analysis indicated it had a 26aa signal peptide and was a secretory
pathway protein. RT-PCR demonstrated that the expression of PoIR2 was quickly induced and drastically increased in liver, kidney, spleen, gills, intestine, heart, and skeletal muscle after
infected with V. anguillarum. These results indicated that the PoIR2 might play some important role in Japanese flounder immune response system. This gene was named PoIR2 (P.olivaceus immune-related gene 2, GenBank accession number: EU224372). The mature PoIR2 peptide was expressed in BL21(DE3) pLysS using pET-32a(+) vector and a great part of the recombinant mature peptide existed
as soluble type. 相似文献
9.
《中国海洋大学学报(英文版)》2021,(4)
The swimming crab Portunus trituberculatus is an economically important marine crustacean species. Here isobaric tags for relative and absolute quantitation(iTRAQ) analysis were used to identify proteins that are differentially expressed during larval development of P. trituberculatus to elucidate the underlying mechanisms. In comparison with the first zoea stage(Z1), 3980 proteins were identified from 32789 peptides, which were matched with 115497 spectrums. A total of 241 proteins were screened with significantly differential expressions in all development stages. These 241 proteins are involved in various biological processes, such as cytoskeleton organization, protein synthesis, energy production and substance metabolism, physiological activities, and transport.Cluster analyses of the 241 differentially expressed proteins led to the generation of four protein clusters based on the overall similarity in protein expression patterns. Exactly 54, 70, 36, and 45 proteins clustered in profiles 10(0, 0, 0,-1, 0, 0), 15(0, 1, 0, 1, 0, 1), 18(0, 1, 2, 2, 1, 0), and 19(0, 1, 2, 3, 4, 5), respectively. Muscle development and exoskeleton renewal were important processes throughout the development stages. In addition, protein synthesis, degradation, and digestion actively occurred, especially at the Z4 stage. These results provide novel insights into the mechanisms underlying larval development of swimming crab and can assist in larval rearing. 相似文献
10.
Zheng Jianlu Wang Zhaoding Lin Zhiqing Li Zijiang Zhu Zhuohong Chen Jinsi 《中国海洋湖沼学报》1984,2(2):133-140
Nine chemical species of Zn, Cd, Pb and Cu in the Zhujiang River estuary were measured by using the technique of ion-exchange,
ultraviolet irradiation and anodic stripping voltammetry. According to the distribution of the nine species in percentage
an estuarine chemical pattern of the metal elements Zn, Cd, Pb and Cu in the Zhujiang River estuary was roughly made, and
its is suggested that this estuary has relatively great self purifying ability in respect to heavy metal pollution.
This paper was published in Chinese in theOceanologia et limnologia sinica
13(1): 19–25, 1983. 相似文献
11.
12.
《中国海洋湖沼学报》2016,(2)
Members of the Dna J family are proteins that play a pivotal role in various cellular processes, such as protein folding, protein transport and cellular responses to stress. In the present study, we identified and characterized the full-length Dna J c DNA sequence from expressed sequence tags of Pyropia yezoensis( Py Dna J) via rapid identification of c DNA ends. This c DNA encoded a protein of 429 amino acids, which shared high sequence similarity with other identified Dna J proteins, such as a heat shock protein 40/Dna J from Pyropia haitanensis. The relative m RNA expression level of Py Dna J was investigated using real-time PCR to determine its specific expression during the algal life cycle and during desiccation. The relative m RNA expression level in sporophytes was higher than that in gametophytes and significantly increased during the whole desiccation process. These results indicate that Py Dna J is an authentic member of the Dna J family in plants and red algae and might play a pivotal role in mitigating damage to P. yezoensis during desiccation. 相似文献
13.
To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii, a high efficient expression vector was constructed. Green fluorescent protein （GFP） was expressed in C. reinhardtii under the control of promoters： RBCS2 and HSP70A-RBCS2. Efficiency of transformation and expression were compared between two transgenic algae： RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ. Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2, and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ. In addition, a threefold increase of GFP in Tran-Ⅱ was induced by heat shock at 40℃. All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C. reinhardtii. 相似文献
14.
Insulin-like growth factors (IGFs) are key regulators of development and growth. Here, we characterized the igf2 gene from olive flounder (Paralichthys olivaceus) and determined its temporal and spatial expression. We set up an in-vitro protein expression system in eukaryotic human embryonic kidney (HEK293T) cells and explored its effects on cell proliferation. The flounder igf2 cDNA contained a 648-bp open reading frame (ORF) encoding a protein of 215 amino acids (aa), which spanned the complete signal peptide (47 aa), mature peptide (70 aa), and E domain (98 aa). In adult flounder, igf2 mRNA was detected in all selected tissues. In early development, igf2 mRNA was detected throughout development from unfertilized eggs to hatching-stage embryos. In-situ hybridization analysis indicated that igf2 mRNA was specially expressed in the brain region, floor plate, hypochord, otic vesicle, and pectoral fin during embryogenesis. Western blotting analysis indicated that the soluble recombinant flounder IGF2 protein was successfully produced through eukaryotic expression in HEK293T cells. In addition, the recombinant IGF2 protein significantly promoted the proliferation of human cervical carcinoma (HeLa) and HEK293T cells. These results provide new information about the structural and functional conservation, expression patterns, and biological activity of the igf2 in teleosts.
相似文献15.
16.
Expression of putative zinc-finger protein lcn61 gene in lymphocystis disease virus China （LCDV-cn） genome 总被引：1，自引：0，他引：1
An open reading frame (lcn61) of lymphocystis disease virus China (LCDV-cn), probably responsible for encoding putative zinc-finger proteins was amplified
and inserted into pET24a (+) vector. Then it expressed in E. coli BL21 (DE3), and His-tag fusion protein of high yield was obtained. It was found that the fusion protein existed in E. coli mainly as inclusion bodies. The bioinformatics analysis indicates that LCN61 is C2H2 type zinc-finger protein containing
four C2H2 zinc-finger motifs. This work provides a theory for functional research of lcn61 gene.
Supported by High Technology Research and Development Program of China (863 Program, No. 2006AA100309) 相似文献
17.
18.
The Gram-negative Vibrio parahaemolyticus is a common pathogen in humans and marine animals. Bacteria flagellins play an important role during infection and induction
of the host immune response. Thus, flagellin proteins are an ideal target for vaccines. We amplified the complete flagellin
subunit gene (flaA) from V. parahaemolyticus ATCC 17802. We then cloned and expressed the gene into Escherichia coli BL21 (DE3) cells. The gene coded for a protein that was 62.78 kDa. We purified and characterized the protein using Ni-NTA
affinity chromatography and Anti-His antibody Western blotting, respectively. Our results provide a basis for further studies
into the utility of the FlaA protein as a vaccine candidate against infection by Vibrio parahaemolyticus. In addition, the purified FlaA protein can be used for further functional and structural studies. 相似文献
19.
20.
Liao Yulin 《中国海洋湖沼学报》1989,7(4):339-344
Two new species of the ophiuriod genusAmphilimna from 280 m depth water east of Hainan Island are described.A. sinica sp. nov. shows affinities withA. transacta (Koehler), andA. granulosa is related toA. cribriformis A. M. Clark.
Contribution No. 1432 from the Institute of Oceanology, Academia Sinica. 相似文献