三叶斑潜蝇热激蛋白Hsp90基因的克隆及其在高温胁迫下的表达分析

采用RT-PCR及RACE技术克隆了三叶斑潜蝇Hsp90基因全长cDNA序列,并用实时定量RT-PCR的方法检测其在不同发育阶段受到高温胁迫后的表达水平。该基因的cDNA序列全长2 408 bp,开放阅读框为2 145 bp,编码714个氨基酸;5′非编码区为151 bp,3′非编码区为112 bp。该基因推导的氨基酸序列与其他昆虫同源序列比较有很高的相似性（80%~99%）。聚类分析结果显示三叶斑潜蝇与美洲斑潜蝇和南美斑潜蝇的亲缘关系最近。实时荧光定量PCR检测结果表明三叶斑潜蝇Hsp90基因的表达受到热胁迫的诱导,诱导3龄幼虫最大表达量的温度比诱导其他发育阶段的温度低,在43 ℃时预蛹和蛹的表达量在整个生命周期中最高,在检测的高温胁迫条件下,雄虫比雌虫的表达量更高。该结果为阐明三叶斑潜蝇胁迫耐受能力及其对其他潜蝇种群的取代机制奠定了分子基础。     

60Co-γ辐射对茭白叶片光合特性和荧光参数的影响

分别以50,75和100Gy剂量水平的60Co-γ射线对3叶期的茭白(Zizania latifolia)幼苗进行辐射处理,通过测定其叶绿素含量、光合气体交换指标和叶绿素荧光参数来探讨γ射线辐射对茭白植株光合和荧光生理特性的影响.试验结果表明,75 Gy剂量处理茭白的叶绿素a、叶绿素b和总叶绿素含量降低显著;不同剂量的...     

Study on Several Characteristics of Agaricus blazei Murril Strain J3 Irradiated by 60Co

A new mutant strain J3 from Agaricus blazei Murril was obtained by 60Co irradiation. The yields of successive generations were increased more than 70% to compare with their original strain. The component analysis on amino acids and fatty acid illuminated that the nutrient value of strain J3 fruiting body was better than the original strain. The apparent nodule structure was found in the hyphea of J3 strain through the observation by scanning electron microscope. RAPD analysis showed the great difference of PCR fingerprints between J3 and its original strain. It is a promising mutant strain for further commercial development in the future.     

~（60）Co辐照装置机电设备和安全联锁系统的设计

设计了￣（６０）Ｃｏ辐照装置的机械设备、控制设备和安全联锁系统。并分别介绍了上述三部分的工作原理、技术关键和系统性能，最后通过运行试验，证明该装置满足设计要求。     

兔出血症病毒VP60蛋白单克隆抗体的制备及活性鉴定

为制备兔出血症病毒VP60单克隆抗体,利用HEK293T细胞瞬时表达VP60,免疫Balb/c小鼠,将小鼠脾细胞与SP2/0骨髓瘤细胞融合,间接ELISA筛选获得阳性克隆细胞株-5D11。利用血凝抑制(HI)、间接免疫荧光(IFA)和Westernblot等方法对5D11分泌的单克隆抗体活性进行鉴定,并对其识别区域进行了定位。结果显示,5D11能抑制病毒凝集人“O”型红细胞,与杆状病毒表达的VP60蛋白发生特异性抗原抗体反应,且其识别VP60蛋白线性抗原表位,并定位于494aa-579aa区段。成功制备了抗VP60蛋白单克隆抗体,为开展病原学检测、流行病学研究等奠定了基础。     

冷暴露仔猪PBMC中Hsp72与血浆TNF-α、IL-6的变化规律

为探讨急性冷应激对仔猪免疫系统的影响及机制,本试验将18头仔猪随机分成常温对照组、冷暴露组和冷暴露并糖皮质激素受体(GRs)阻断组.常温组在18℃～20℃下而冷暴露各组在4℃～8℃环境下分栏饲养.试验期持续12 h,在冷暴露开始后六个时间点对18头仔猪采血并分离血浆和外周血单个核细胞(PBMC),检测Hsp72和TNF-α、IL-6的水平.实验结果显示:冷暴露组IL-6水平在冷暴露6 h时比开始前有显著升高(P＜0.01),TNFα水平在冷暴露1 h 时显著下降(P＜0.01),3 h和6 h又显著升高(P＜0.01),且IL-6与TNF-α呈中等相关(r=0.4198);GRs阻断组IL-6水平在冷暴露6 h时显著升高,TNF-α水平在各时间点差异不显著;冷暴露各组Hsp72在冷暴露1 h,3 h,6 h,9 h,12 h时均显著升高(P＜0.01),且受体阻断组TNF-α与Hsp72呈中等强度负相关(r=0.6444).这些结果表明:急性冷暴露能使血浆IL-6水平升高和PBMC中Hsp72 的表达增强,而使TNF-α的水平降低,且阻断糖皮质激素对它们之间的相关性有一定影响.     

马立克病肿瘤组织内HSP60表达及生物学作用初步研究

本试验旨在研究HSP60与马立克病肿瘤发生、发展之间的相关性。通过人工感染,建立鸡马立克病肿瘤模型,定期剖杀,利用病理组织学和免疫组织化学方法,检测HSP60与肿瘤细胞定位之间的相关性;设计HSP60 RNA干扰序列,构建重组慢病毒,转染MSB-1细胞,利用流式细胞技术,探索降低HSP60转录表达对MSB-1细胞凋亡水平的影响。结果显示：HSP60在肿瘤细胞的细胞质内强表达;成功构建了HSP60 RNA干扰慢病毒,且5147序列干扰效果最佳;5147序列慢病毒转染48 h时,与对照序列组和空白对照组相比,HSP60转录、表达水平极显著降低（P<0.01）,MSB-1细胞凋亡水平极显著升高（P<0.01）。在马立克病肿瘤发生、发展过程中,HSP60组织细胞定位与肿瘤细胞具有明显的相关性,降低HSP60表达水平能够导致MSB-1细胞凋亡升高,说明HSP60对肿瘤细胞的存活具有重要的生物学作用。     

姬松茸60Co辐射诱变菌株J5与原菌株J1蛋白质

采用非生物学评价法对姬松茸60Co辐射诱变菌株J5和其原菌株J1不同潮次菇蛋白质的营养价值进行评价.结果表明:J5子实体的必需氨基酸指数、生物价、营养指数、氨基酸比值系数分、化学评分和氨基酸评分分别比J1高1.02％、2.26％、2.93％、11.72％、9.35％和5.13％；以上6项蛋白质评价指标中,J5第1、第2...     

cPKCγ-HSP60 signal pathway participates in hypoxic preconditioning in mice

AIM: To identify the proteins interacted with conventional protein kinase Cγ (cPKCγ) which are involved in hypoxic preconditioning (HPC), and to investigate the role of cPKCγ-interacted heat-shock protein 60 (HSP60) in the development of HPC and ischemia (I).METHODS: Healthy male BALB/c mice were randomly divided into normoxia(Norm) and HPC groups. Based on whether middle cerebral artery occlusion (MCAO) was performed, the mice were divided into Norm+sham group, Norm+I group, HPC+sham group and HPC+I group ( all n=6). Immunoprecipitation, two-dimensional electrophoresis and mass spectrometry were applied to identify the proteins interacted with cPKCγ. The changes of HSP60 expression in the brain of the mice after HPC and MCAO were analyzed by Western blotting.RESULTS: The interaction of cPKCγ and HSP60 was confirmed by co-immunoprecipitation result. Compared with Norm groups, the expression level of cPKCγ-interacted HSP60 obviously increased in particulate fraction of cerebral cortex in HPC mice. In the MCAO ischemic animals, the level of HSP60 expression was significantly higher in ischemic core and penumbra in Norm+I group and HPC+I group than that in Norm+sham group. HSP60 expression in ischemic core was lower in HPC+I group than that in Norm+I group.CONCLUSION: cPKCγ-HSP60 signal pathway might be involved in the development of cerebral hypoxic preconditioning in MCAO mice.