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Reversible protein phosphorylation, catalyzed by protein kinases and phosphatases, is an important and versatile mechanism by which eukaryotic cells regulate almost all the signaling processes. Protein phosphatase 1 (PP1) is the first and well-characterized member of the protein serine/threonine phosphatase family. In the present study, a full-length cDNA encoding the beta isoform of the catalytic subunit of protein phosphatase l(PPlcb), was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus, designated SmPPlcb, by the rapid amplification of cDNA ends (RACE) technique. The cDNA sequence of SmPPlcb we obtained contains a 984 bp open reading frame (ORF), flanked by a complete 39 bp 5' untranslated region and 462 bp 3' untranslated region. The ORF encodes a putative 327 amino acid protein, and the N-terminal section of this protein is highly acidic, Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp, a common feature for PP1 catalytic subunit but absent in protein phosphatase 2B (PP2B). And its calculated molecular mass is 37 193 Da and pI 5.8. Sequence analysis indicated that, SmPPlcb is extremely conserved in both amino acid and nucleotide acid levels compared with the PPlcb of other vertebrates and invertebrates, and its Kozak motif contained in the 5'UTR around ATG start codon is GXXAXXGXXATGG, which is different from mammalian in two positions A6 and G3, indicating the possibility of different initiation of translation in turbot, and also the 3'UTR of SmPPlcb is highly diverse in the sequence similarity and length compared with other animals, especially zebraf'lsh. The cloning and sequencing of SmPPlcb gene lays a good foundation for the future work on the biological functions of PP1 in the flatfish turbot. 相似文献
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miRNA(microRNA)是一类广泛存在的长度约为18~25nt(nucleotides)的单链非编码小RNA。它在转录后水平通过降解靶基因的mRNA或者抑制靶基因mRNA的翻译来调控靶基因的表达,在生物的生长、发育、代谢和免疫等多种生命活动中发挥重要作用。对虾miRNA的研究起步较晚,但近6年来取得了突飞猛进的进展。本文综述了对虾miRNA的发现与鉴定、作用机制及其生物学功能方面的研究概况,并对对虾miRNA的研究前景进行了展望。 相似文献
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利用牙鲆鳃细胞系分离和培养淋巴囊肿病毒 总被引:1,自引:4,他引:1
本文利用牙鲆鳃细胞系进行了养殖牙鲆淋巴囊肿病毒的分离及培养 ,并通过电镜对培养细胞中淋巴囊肿病毒的形态及感染循环进行了初步研究。将病鱼的淋巴囊肿组织无菌滤液接种牙鲆细胞系 ,细胞出现了明显的细胞病变 ( Cytopathic effect,CPE)。电镜观察在培养细胞的胞质中有病毒的包涵体 ,胞质中散在 6角形、5角形或圆形的病毒粒子 ,大小为 10 0~ 140 nm之间。在感染细胞的线粒体中也存在大量的病毒颗粒。 相似文献
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水生动物的细胞培养晚于陆生动物 ,其中 (包括水生无脊椎动物和水生脊椎动物 ) ,水生无脊椎动物的细胞培养主要是甲壳类 (如对虾、螯虾、龙虾、蟹和鲎 )、贝类 (如牡蛎、珍珠贝和蛤 )和海绵的体外培养。水生无脊椎动物的细胞培养比较困难 ,原代培养细胞不能形成单层 ,或即使形成单层 ,但细胞不分裂 ,难以进行传代。因而 ,至今没有得到水生无脊椎动物的连续性细胞系。水生无脊椎动物的细胞培养不仅在甲壳类和贝类的病毒分离、检测和疫苗制备方面有很大的应用价值 ,而且还可能用来生产天然产物比如药物。已经发现不少海洋无脊椎动物 (如海绵、… 相似文献
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用16S rRNA基因的内切酶图谱快速鉴别几种对虾病原菌 总被引:4,自引:0,他引:4
坎普氏弧菌是青岛海洋大学生物系微生物实验室于1989-1990年自对虾养殖场中国对虾红腿病心脏及血淋巴中分离并鉴定的菌株,副溶血菌和溶藻胶弧菌两菌株于1994年9月得自中国科学院微生物研究所,为建立快速、准确的中国对虾病原菌的诊断技术,根据几种细菌的16SrRNA基因的序列,设计并合成该基因的多聚酶反应的引物PL1和PL2。并用该对引物分别从坎普氏弧菌、副溶血弧菌和溶藻胶弧菌的DNA要品中扩增出分 相似文献
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Three continuous marine fish cell lines of FG (i. e., Hounder Gill) from flounder (Paralichthys olivaceus) gill, SPH (i. e. , Sea Perch Heart) from sea perch (Lateolabrax japonicus) heart and RSBF (i.e., Red Sea Bream Fin) from red sea bream (Pagrosomus major) fin, were characterized by lactate dehydrogenase (LDH) isozyme and morphological analysis. The LDH isozyme patterns of these three cell lines and their corresponding tissues of origin were investigated and compared. The results showed: (1) No difference was found in the LDH isozyme patterns of FG and flounder gill tissue. However, the LDH isozyme patterns of SPH and RSBF were significantly different from their corresponding tissues of origin; (2) LDH isozyme patterns of FG, SPH and RSBF were markedly different from each other and could serve as genetic markers for species identification and detection of cross contamination. Morphological change analysis of these three cell lines in comparison to their original tissues indicated that FG cells still appeared epithelioid without morphological transformation. However, morphological changes were found in SPH and RSBF compared to their original tissues. Therefore, the cellular morphology was still plastic in the relatively stable culture conditions, and it was possible that change of LDH patterns wasrelated to morphological changes of fish cells in vitro. 相似文献
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CRISPR/Cas9(Clustered regularly interspaced short palindromic repeat/CRISPR-associated nuclease 9)技术是一种由gRNA(guide RNA)引导的Cas9核酸内切酶靶向编辑技术,已广泛用于基因的敲除、敲入和敲降,其操作简便快速,成本低,已成为人们探究基因功能、修复受损基因、沉默有害基因以及改良经济物种种质性状的重要工具。目前,该技术已经在少数几种水生甲壳动物中得到成功应用,但是由于显微注射技术在应用上的局限性,该技术在许多重要海水养殖经济物种如对虾中的应用受到限制,导致其基因编辑效率低下,难以广泛开展。本文对CRISPR/Cas9基因编辑技术在水生甲壳动物中的应用进行了综述和展望,为今后海水养殖虾蟹类的基因功能研究以及遗传育种研究提供参考。 相似文献
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将经直接消化的斑马鱼头肾细胞或经短暂培养的尾鳍细胞核移植到同种成熟具核卵子中 ,移核卵可发育到原肠晚期 ;将移核卵发育成的囊胚细胞核作供体进行第 2次核移植 ,受体卵可发育到尾芽期或肌肉感应期。流式细胞仪测定 DNA含量表明 ,受体卵发育形成的囊胚细胞 DNA含量与正常囊胚细胞 DNA含量相同 ,由此可知 ,移核卵中的雌原核没有参与移核胚胎的发育。 相似文献