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HAO Feng BAI Xue-song JU Xiao-hong FANG Fang ZANG Yu-xuan ZHU Hang-fei XIANG Guo-yan ZHANG Yun-qiao YUAN Zhong-hai 《园艺学报》2014,30(9):1633-1639
AIM:To investigate the expression of transmembrane protein 16A(TMEM16A) in Fischer rat thyroid follicular epithelial (FRT) cells and its electrophysiologic properties. METHODS:The eukaryotic expression vector of pUB6/V5-TMEM16A was constructed and transfected into FRT cells by liposome-mediated transfection. In order to obtain the high efficiency of gene transfection and expression, the quantity and ratio of lipid/DNA complexes were optimized. The FRT cells stably expressing TMEM16A were gained by the selection with blasticidin and confirmed by the techniques of RT-PCR and immunofluorescence. The expression and location of TMEM16A in the FRT cells were observed under an inverted fluorescence microscope. TMEM16A protein was associated with calcium-dependent chloride current, as measured with halide-sensitive fluorescent protein and patch-clamp technique. RESULTS:The results of double digestion and sequencing indicated that TMEM16A was cloned into pUB6/V5. The results of RT-PCR and immunofluorescence confirmed that TMEM16A was expressed in the FRT cells after transfection with TMEM16A. The classical calcium-activated chloride channel currents were recorded in the FRT cells stably expressing TMEM16A by the technique of patch-clamp and halide-sensitive fluorescent protein YFP-H148Q/I152L. CONCLUSION:The protein expression of TMEM16A in the FRT cells was observed. TMEM16A is the molecular identity of calcium-activated chloride channels. 相似文献
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长肢秀体溞对富营养化水体藻类的生物操纵 总被引:8,自引:0,他引:8
在实验室和开放生态系统中研究了长肢秀体溞(D iaphanosoma leuchtenbergianumF ischer)对斜生栅藻(Scem edesmus obliquusTurp)、铜绿微囊藻(M icrocystis aeruginosa)和蛋白核小球藻(Chlorella pyrenoidosaCh ick)的牧食力和对水体中浮游动物和藻类数量、生物量、群落结构及水体中理化性质的影响,结果表明:藻类生物量在一定范围内,长肢秀体溞的摄食速度随藻类生物量增加而增加,藻类生物量超过一定值(即饱和值)时,摄食速度随之下降。长肢秀体溞对斜生栅藻和小球藻具有较高的选择性,对铜绿微囊藻的选择性较差。水泥池中每升水放养长肢秀体溞600个以上,即可对水体中浮游动物和藻类的数量、生物量、群落结构产生显著的影响,同时降低水体中总氮、总磷和CODMn的浓度,增加水体透明。7月份之前,可以利用放养长肢秀体溞的方法来控制水体中藻类。 相似文献
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以带鱼为原料,通过双酶分步水解制备蛋白酶解液,利用活性炭吸附脱除芳香族氨基酸,紫外和氨基酸组成分析测定高F值寡肽(HFP)的F值,并对昆明小鼠体内的活性进行了系统评价。酶解实验结果显示,带鱼蛋白采用胃蛋白酶和风味蛋白酶进行双酶分步水解,胃蛋白酶的最优酶解工艺参数为酶用量3%,料液比1∶5(g/m L),温度35°C,p H 2.0,酶解时间3 h;风味蛋白酶的最优酶解工艺参数为酶用量3%,p H 7.0,温度50°C,酶解时间3 h。活性炭型号筛选和脱芳工艺研究表明,4号粉末活性炭(200目)优于其他4种活性炭,在料液比1∶20(g/m L)、p H 6.0、常温下处理4 h,蛋液的脱芳率最高,F值(OD220/OD280)由脱芳前的5.75增加到脱芳后的30.72。制备的带鱼高F值寡肽经氨基酸组成分析验证,F值(支链氨基酸/芳香族氨基酸)为28.06。活性实验结果显示,与对照组相比,高F值寡肽剂量组昆明小鼠的醉酒时间明显延长,醒酒时间显著缩短,力竭游泳时间明显延长,游泳后的剂量组肝糖原和肌糖原含量增加,血清尿素氮和乳酸含量下降,而乳酸脱氢酶活性则明显提高。另外,剂量组小鼠体内的抗氧化酶SOD、GSH-Px和CAT的活性均显著提高,而丙二醛含量则显著减少。本实验为带鱼高F值寡肽的制备提供技术支持,所制备的高F值寡肽对小鼠具有较好的抗醉酒、醒酒、抗疲劳作用和抗氧化活性。 相似文献
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收集鱼鳞云杉健康木和被害木干部挥发性物质,利用GC-MS方法对挥发性物质进行定性和定量分析,比较两者之间挥发性物质种类和含量的差异,分析鱼鳞云杉干部挥发性成分与云杉大黑天牛寄主选择的关系。结果表明:健康鱼鳞云杉干部挥发性物质共42种成分,其中相对质量分数最大的是神圣亚麻三烯(16.04%),其次是α-葑烯(14.24%);被害鱼鳞云杉干部挥发性物质共43种,其中相对质量分数最大的成分是β-蒎烯(18.07%),其次是α-蒎烯(12.39%)。在健康木和被害木的挥发性成分中,烯萜类化合物均占70%以上。鱼鳞云杉树势由健康向衰弱转变时,主要烯萜类化合物含量发生变化,3-蒈烯、异柠檬烯、β-蒎烯、4-蒈烯、α-蒎烯等含量显著上升,而神圣亚麻三烯、α-葑烯、β-月桂烯、β-水芹烯等含量下降,这些烯萜类化合物的变化,为云杉大黑天牛定位选择适宜寄生的鱼鳞云杉提供了依据。 相似文献
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Tada Y Yano N Takahashi H Yuzawa K Ando H Kubo Y Nagasawa A Chin K Kawamata Y Sakai R Ohashi N Ogata A Nakae D 《Journal of toxicologic pathology》2010,23(1):39-47
A subchronic feeding study of l-serine (l-Ser) was conducted with groups of 10 male and 10 female Fischer 344 rats fed a powder diet containing 0, 0.06, 0.5, 1.5 or 5.0% concentrations of l-Ser for 90 days. There were no toxicologically significant, treatment-related changes with regards to body weight, food intake, water intake or urinalysis data. In several of the hematology, serum biochemistry and organ weight parameters, significant changes were observed between some of the treated groups and the controls. All these changes, however, were subtle and lacked any corresponding pathological findings. In addition, the increased or decreased values remained within the range of the historical control values. In fact, histopathological assessment revealed only sporadic and/or spontaneous lesions. In conclusion, the no-observed-adverse-effect-level (NOAEL) for l-Ser was, therefore, determined to be at least a dietary dose of 5.0% (2765.0 mg/kg body weight/day for males and 2905.1 mg/kg body weight/day for females) under the present experimental conditions. 相似文献
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[目的]探讨更为客观的木霉菌鉴定手段。[方法]在形态分类的基础上结合分子鉴定手段(rDNA-ITS序列分析),对3株茶藨生柱锈重寄生木霉菌进行分类鉴定。[结果]依据TR1的培养性状和显微特征描述,初步鉴定该菌株为深绿木霉(T.atroviride);TR2、TR3的培养性状和显微特征在一定程度上相似,依据其形态特征,初步鉴定这2个菌株为绿色木霉(T.viridePers.ex Fr.)。从Genbank中深绿木霉和绿色木霉的6个菌株与TR1、TR2T、R3所作的系统发育树可知,TR1应该归为深绿木霉,TR2和TR3属同种真菌,应该归为绿色木霉,这与形态学观察结果一致。[结论]形态特征结合ITS序列分析可作为木霉菌分类鉴定的依据。 相似文献