The basic genetic characteristics, important functional genes, and entire transcriptome of Solen grandis Dunker were investigated by constructing a full-length cDNA library with the ‘switching mechanism at the 5'...The basic genetic characteristics, important functional genes, and entire transcriptome of Solen grandis Dunker were investigated by constructing a full-length cDNA library with the ‘switching mechanism at the 5'-end of the RNA transcript'(SMART) technique. Total RNA was isolated from the immune-relevant tissues, gills and hemocytes, using the Trizol reagent, and cDNA fragments were digested with Sfi I before being ligated to the pBluescript II SK* vector. The cDNA library had a titer of 1048 cfu μL-1 and a storage capacity of 1.05×106 cfu. Approximately 98% of the clones in the library were recombinants, and the fragment lengths of insert cDNA ranged from 0.8 kb to 3.0 kb. A total of 2038 expressed sequence tags were successfully sequenced and clustered into 965 unigenes. BLASTN analysis showed that 240 sequences were highly similar to the known genes(E-value < 1e-5; percent identity >80%), accounting for 25% of the total unigenes. According to the Gene Ontology, these unigenes were related to several biological processes, including cell structure, signal transport, protein synthesis, transcription, energy metabolism, and immunity. Fifteen of the identified sequences were related to defense and immunity. The full-length cDNA sequence of HSC70 was obtained. The cDNA library of S. grandis provided a useful resource for future researches of functional genomics related to stress tolerance, immunity, and other physiological activities.展开更多
Amphioctopus fangsiao is one of the most economically important species and has been considered to be a candidate for aquaculture. In order to facilitate its fine-scale genetic analyses, we constructed a normalized fu...Amphioctopus fangsiao is one of the most economically important species and has been considered to be a candidate for aquaculture. In order to facilitate its fine-scale genetic analyses, we constructed a normalized full-length library successfully and developed a set of microsatellite markers in this study. The normalized full-length library had a storage capacity of 6.9×105 independent clones. The recombination efficiency was 95% and the average size of inserted fragments was longer than 1000 bp. A total of 3440 high quality ESTs were obtained, which were assembled into 1803 unigenes. Of these unigenes, 450(25%) were assigned into 33 Gene Ontology terms, 576(31.9%) into 153 Kyoto Encyclopedia of Genes and Genomes pathways, and 275(15.3%) into 22 Clusters of Orthologous Groups. Seventy-six polymorphic microsatellite markers were identified. The number of alleles per locus ranged from 4 to 17, and the observed and expected heterozygosities varied between 0.167 and 0.967 and between 0.326 and 0.944, respectively. Twelve loci were significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction and no linkage disequilibrium was found between different loci. This study provided not only a useful resource for the isolation of the functional genes, but also a set of informative microsatellites for the assessment of population structure and conservation genetics of A. fangsiao.展开更多
Interleukin-17(IL-17),a prototype member of IL-17 family,plays an important role in defending against extracellular pathogens as a pro-inflammatory cytokine.The function and distribution of IL-17 have been extensively...Interleukin-17(IL-17),a prototype member of IL-17 family,plays an important role in defending against extracellular pathogens as a pro-inflammatory cytokine.The function and distribution of IL-17 have been extensively studied in many vertebrates.However,few study has focused on the role of IL-17 in invertebrates,especially in mollusks.In this study,an IL-17 homolog was identified in Octopus ocellatus,which was designated as OoIL-17.The phylogenetic analysis showed that OoIL-17 is clustered well with other invertebrate IL-17,indicating it is highly similar with the IL-17 of other invertebrates.The expression of OoIL-17 gene was analyzed with qRT-PCR in a variety of healthy tissues and the hemocytes infected with Vibro anguillarum or Micrococcus luteus.The mRNA of OoIL-17 gene is constitutively expressed at different levels in all examined tissues of healthy O.ocellatus,including mantle,stomach,hemocytes,muscle,gonad,hepatopancreas,systemic heart and gill.The lowest expression was observed in mantle while the highest was observed in hepatopancreas.The expression level of OoIL-17 gene is significantly up-regulated in O.ocellatus hemocytes upon infection with V.anguillarum and M.luteus,indicating its active involvement in the host immune response against bacterial pathogens.The results laid the foundation for further understanding the innate immune mechanisms of IL-17 in O.ocellatus and mollusks.展开更多
基金supported by the Shandong Program of Agriculture Thoroughbred Project to Xiangquan Liu,the Marine Special Research Foundation of China for Non-profit Public Industry(No.200805031)Taishan Scholar position funding of Aquatic Animal Nutrition and Feed to Linmin Zhang and NSFC funding(No.31202025)
文摘The basic genetic characteristics, important functional genes, and entire transcriptome of Solen grandis Dunker were investigated by constructing a full-length cDNA library with the ‘switching mechanism at the 5'-end of the RNA transcript'(SMART) technique. Total RNA was isolated from the immune-relevant tissues, gills and hemocytes, using the Trizol reagent, and cDNA fragments were digested with Sfi I before being ligated to the pBluescript II SK* vector. The cDNA library had a titer of 1048 cfu μL-1 and a storage capacity of 1.05×106 cfu. Approximately 98% of the clones in the library were recombinants, and the fragment lengths of insert cDNA ranged from 0.8 kb to 3.0 kb. A total of 2038 expressed sequence tags were successfully sequenced and clustered into 965 unigenes. BLASTN analysis showed that 240 sequences were highly similar to the known genes(E-value < 1e-5; percent identity >80%), accounting for 25% of the total unigenes. According to the Gene Ontology, these unigenes were related to several biological processes, including cell structure, signal transport, protein synthesis, transcription, energy metabolism, and immunity. Fifteen of the identified sequences were related to defense and immunity. The full-length cDNA sequence of HSC70 was obtained. The cDNA library of S. grandis provided a useful resource for future researches of functional genomics related to stress tolerance, immunity, and other physiological activities.
基金the National Natural Science Foundation of China (Nos. 31302215, 31272643)the Shandong Provincial Natural Science Foundation (Nos. BS2014NY010, ZR2013CQ030)the Shandong Provincial Primary Research and Development Projects (No. 2015GNC110017)
文摘Amphioctopus fangsiao is one of the most economically important species and has been considered to be a candidate for aquaculture. In order to facilitate its fine-scale genetic analyses, we constructed a normalized full-length library successfully and developed a set of microsatellite markers in this study. The normalized full-length library had a storage capacity of 6.9×105 independent clones. The recombination efficiency was 95% and the average size of inserted fragments was longer than 1000 bp. A total of 3440 high quality ESTs were obtained, which were assembled into 1803 unigenes. Of these unigenes, 450(25%) were assigned into 33 Gene Ontology terms, 576(31.9%) into 153 Kyoto Encyclopedia of Genes and Genomes pathways, and 275(15.3%) into 22 Clusters of Orthologous Groups. Seventy-six polymorphic microsatellite markers were identified. The number of alleles per locus ranged from 4 to 17, and the observed and expected heterozygosities varied between 0.167 and 0.967 and between 0.326 and 0.944, respectively. Twelve loci were significantly deviated from Hardy-Weinberg equilibrium after Bonferroni correction and no linkage disequilibrium was found between different loci. This study provided not only a useful resource for the isolation of the functional genes, but also a set of informative microsatellites for the assessment of population structure and conservation genetics of A. fangsiao.
基金supported by the earmarked fund for the Modern Agro-Industry Technology Research System (No. CARS-49)the Natural Science Foundation of Shandong Province (No. ZR2019BC052)the Marine and Fisheries Science and Technology Innovation Program of Shan-dong Province (No. 2017YY04)
文摘Interleukin-17(IL-17),a prototype member of IL-17 family,plays an important role in defending against extracellular pathogens as a pro-inflammatory cytokine.The function and distribution of IL-17 have been extensively studied in many vertebrates.However,few study has focused on the role of IL-17 in invertebrates,especially in mollusks.In this study,an IL-17 homolog was identified in Octopus ocellatus,which was designated as OoIL-17.The phylogenetic analysis showed that OoIL-17 is clustered well with other invertebrate IL-17,indicating it is highly similar with the IL-17 of other invertebrates.The expression of OoIL-17 gene was analyzed with qRT-PCR in a variety of healthy tissues and the hemocytes infected with Vibro anguillarum or Micrococcus luteus.The mRNA of OoIL-17 gene is constitutively expressed at different levels in all examined tissues of healthy O.ocellatus,including mantle,stomach,hemocytes,muscle,gonad,hepatopancreas,systemic heart and gill.The lowest expression was observed in mantle while the highest was observed in hepatopancreas.The expression level of OoIL-17 gene is significantly up-regulated in O.ocellatus hemocytes upon infection with V.anguillarum and M.luteus,indicating its active involvement in the host immune response against bacterial pathogens.The results laid the foundation for further understanding the innate immune mechanisms of IL-17 in O.ocellatus and mollusks.
