青蛤(Cyclina sinensis)TLR2基因的克隆与表达分析

利用转录组文库分析得到青蛤(Cyclina sinensis)的Toll样受体2(Toll-like receptor 2,TLR2)基因序列,采用荧光定量PCR法分析该基因的表达过程。结果显示,青蛤TLR2基因的cDNA开放阅读框为2082bp,编码693个氨基酸,具典型的LRRs、单次跨膜区和TIR结构域。该基因在血淋巴中的表达量最高,与肝脏、鳃、外套膜、闭壳肌和性腺有显著性差异(P0.05)。在鳗弧菌和Poly I:C刺激下,青蛤血淋巴中TLR2基因3h开始升高,48h达到最大值,与对照组和空白组有极显著性差异(P0.01);藤黄微球菌刺激下,血淋巴中CsTLR2基因3h开始升高,48h亦达到最大值,实验组与对照组有显著性差异(P0.05)。     

Optimization of the purification methods for recovery of recombinant growth hormone from Paralichthys olivaceus

This study aimed to optimize the purification of recombinant growth hormone from Paralichthys olivaceus. Recombinant flounder growth hormone (r-fGH) was expressed by Escherichia coli in form of inclusion body or as soluble protein under different inducing conditions. The inclusion body was renatured using two recovery methods, i.e., dilution and dialysis. Thereafter, the refolded protein was purified by Glutathione Sepharase 4B affinity chromatography and r-fGH was obtained by cleavage of thrombin. For soluble products, r-fGH was directly purified from the lysates by Glutathione Sepharase 4B affinity chromatography. ELISA-receptor assay demonstrated that despite its low receptor binding activity, the r-fGH purified from refolded inclusion body had a higher yield (2.605 mgL-1) than that from soluble protein (1.964 mgL-1). Of the tested recovery methods, addition of renaturing buffer (pH 8.5) into denatured inclusion body yielded the best recovery rate (17.9%). This work provided an optimized purification method for high recovery of r-fGH, thus contributing to the application of r-fGH to aquaculture.     

斜带石斑鱼(Epinephelus coioides)两种催乳素受体基因的 cDNA 克隆和 mRNA 表达分析

采用RT-PCR方法克隆了斜带石斑鱼两种催乳素受体(Prolactin receptor,PRLR)的cDNA序列,序列分析表明:PRLR1开放阅读框为1947bp,共编码649个氨基酸,PRLR2开放阅读框为1749bp,共编码487个氨基酸,PRLR1与PRLR2的氨基酸同源性为40.4%。用Real-time RT-PCR方法研究了PRLR1和PRLR2在各组织和早期发育不同阶段的表达情况,结果表明:PRLR1和PRLR2在所检测的12种组织中均有表达,其中以鳃、肾、肠表达量较高;PRLR1在受精期表达最高,PRLR2在视囊形成期表达最高,而且除受精卵期外PRLR2在各时期的表达量均高于PRLR1。     

生长激素与胰岛素样生长因子-Ⅰ对马氏珠母贝壳生长形成相关基因表达的影响

研究了外源生长激素与胰岛素样生长因子-Ⅰ处理对马氏珠母贝Pinctada fucata Gould胰岛素相关多肽受体基因(irr)和5种壳基质蛋白基因(nacrein、efcbp、n19、aspein及accbp)表达水平的影响。结果表明,激素处理显著提高了irr基因的表达水平(P<0.05),这与其在软体动物中作为胰岛素样生长因子受体的作用相一致。nacrein基因的表达水平在激素处理组也有显著升高(P<0.05),表明生长激素与胰岛素样生长因子-Ⅰ两种外源激素都增强了马氏珠母贝的生长代谢水平;与对照组相比,n19、aspein与accbp3个基因的表达水平在激素处理组均下调(P<0.05),说明这3个基因的表达受到激素调节通路的抑制作用。此外,研究发现aspein与accbp两个基因的表达在各个实验样本中具有极高的相关性,说明这两个基因在激素通路中可能受到同一个上游因子调控。efcbp基因表达水平在激素处理组与对照组之间表达稳定,各样本之间无显著性的变化(P>0.05)。     

Photocatalytic Degradation of 17α‐Ethinylestradiol and Inactivation of Escherichia coli Using Ag‐Modified TiO2 Nanotube Arrays

Ag‐modified TiO₂ nanotube arrays (Ag/TiO₂ NAs) were prepared and employed as a photocatalyst for degradation of 17α‐ethinylestradiol (EE2) and inactivation of Escherichia coli. The as‐synthesized Ag/TiO₂ NAs were characterized by field‐emission scanning electron microscope (FESEM), X‐ray diffraction (XRD), and X‐ray photoelectron spectroscopy (XPS). It was found that metallic Ag nanoparticles were firmly deposited on the TiO₂ NAs with the pore diameter of 100 nm and the length of 550 nm. Photocatalytic degradation of EE2 and inactivation of E. coli were enhanced effectively in an analogical trend using Ag/TiO₂ NAs. In particular, Ag/TiO₂ NAs exhibited the antimicrobial activity even in the absence of light. The Ag acted as a disinfection agent as well as the dopant of the modified TiO₂ NAs photocatalysis by forming a Schottky barrier on the surface of TiO₂ NAs. Inorganic ions suppressed the rates of photocatalytic degradation of EE2, with HCO having a more pronounced effect than NO or SO. Humic acid (HA) was found to increase the rate of EE2 degradation.     

Potential Atmospheric Transport Pathways for Mercury Measured in the Canadian High Arctic

Large-scale transport pathways for atmospheric gaseous mercury emitted from surrounding source regions to reach the Alert sampling site located at the Northwest Territories of Canada between August 1992 and August 1993 were investigated. The 12-month atmospheric mercury data set comprises blank-corrected total gaseous mercury (TGM) concentrations collected on gold-coated sand traps during one-week long sampling intervals. For TGM concentrations above the 12-month mean TGM level, the analysis reveals regions in Eastern Europe (close to East Germany and Poland), areas on Northeastern America in Canada (Northwest Territories), and central Siberia of the former U.S.S.R. Identification of the physical locations of the emission sources was limited by the temporal resolution of weekly averaged data; however, major atmospheric pathways of mercury transport to the Arctic were successfully resolved. The objective of this note is to demonstrate that the potential source contribution function could be applied to resolve the sources and recover transport pathways of atmospheric mercury over a large scale.     

鱼类维生素D3代谢及其生理功能的研究进展

维生素D(VD)是一类具有抗佝偻病作用的类固醇激素,其中维生素D3(VD3)是VD在动物体内的主要存在形式.作为生命活动必不可少的一类物质,VD发挥着广泛且重要的生理学作用.VD在动物体内的代谢过程及相关的代谢酶具有较高的进化保守性,同时也受到多种因素的调控影响.由于鱼类在生活环境方面与陆生脊椎动物存在显著差异,VD在...     

Analysis of IL-2-1ike factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood, spleen and head kidney of olive flounder,Paralichthys olivaceus. The quantities of IL-2-like factor in the supernatant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes (P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2, (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74±0.67%. This work was supported by National “973” Project G1999012003, G19999012006.