烟夜蛾滞育激素基因在粟酒裂殖酵母中的表达

利用设计的特异性引物通过RT-PCR的方法获得了烟夜蛾滞育激素(Diapause hormone,DH)基因,构建了真核表达栽体pESP-2-DH,并在粟酒裂殖酵母SP-Q01中进行了诱导表达,对表达产物进行了SDS-PAGE电泳,获得了与谷胱甘肽S-转移酶(Glutathione S-tranferases,GST)融合表达的蛋白,分子量约48 kDa,与预测的融合蛋白分子量大小相当.对表达的融合蛋白进行Western blotting检测,结果表明,与GST抗体产生了很强的交叉反应,表明融合蛋白得到了表达.     

H5N1亚型禽流感病毒神经氨酸酶基因的表达及纯化

参考GenBank上发表的H5亚型禽流感病毒(AIV)神经氨酸酶(NA)基因序列设计引物,PCR扩增NA基因,再将其克隆到原核表达载体pET-32a中,用E.coli BL21(DE3)原核表达,SDS-PAGE和Western-blot对表达蛋白进行鉴定.Western-blot结果表明,表达产物具有免疫学活性,蛋白的分子量约为61 kDa,位于包涵体中.包涵体经变性、复性处理,表达蛋白能与H5亚型AIV阳性血清发生特异性反应,具有良好的抗原性.ELISA检测结果表明,用此纯化蛋白作为包被抗原检测H5N1亚型AIV神经氨酸酶抗体具有良好的灵敏性.     

甲基磺酸乙酯对草莓愈伤组织的生理效应

用不同浓度甲基磺酸乙酯对草莓愈伤组织进行诱变处理,测定了愈伤组织中的保护酶和氧化酶活性、总酚含量、MDA含量的变化以及愈伤组织的死亡率。结果表明,随着EMS处理浓度的增加和时间的延长,愈伤组织死亡率呈上升趋势;随着EMS处理浓度的增加,保护酶SOD,CAT和氧化酶PPO,POD活性升高,而后快速下降;随着处理时间的延长,SOD活性逐渐下降,CAT,PPO活性先上升后下降,POD活性则表现为先降后升再降;随着处理浓度的增加和时间的延长,总酚含量变化为先升后降,但上升和下降都比较缓慢,而MDA含量则逐渐上升;处理后的愈伤组织在培养过程中,PPO,POD活性迅速下降并维持在相对稳定的水平,总酚和MDA含量逐渐下降,而总酚含量与PPO,POD活性呈显著正相关。由此认为,在用EMS对草莓愈伤组织进行诱变时,较适宜的剂量为0.1%~0.2%浓度处理1.0~1.5 h,在处理完成后的前12 h培养中,应采取措施以减轻褐变。     

木粒--SPDA的制作与探讨

木粒--SPDA是我们根据木腐生性食用菌在自然状态下,均发生于树木上的特性,以及食用菌在营养需求上与其习性相关的原理,在SPDA基础上添加适量的木粒制成的一种母种培养基.在我们多年的使用中发现,用该培养基培养的母种生长势好,在转接原种时,适应能力强,萌发定植快;用作菌种保藏基质时,保藏时间长,且能很好保持菌种的优良性状.     

Expression of phenylalanine ammonia-lyase cDNA from rice in E. coli BL21DE3

AIM: To study the expression and its kinetics of rice phenylalanine ammonia-lyase gene encoding into E. coli as the basis of treatment for phenylketouria. METHODS: The phenylalanine ammonia-lyase-1-cDNA(rPAL-1-cDNA) from rice was recombined into E. coli high expression vector pET-28c and transformed into E. coli host strain BL21DE3. Engineering bacteria was then inducted by isopropyl-β-D-thiogalactoside (IPTG) for 1, 3, 5, 7 hours, in order to obtain high level expression. RESULTS: After induction, the expression level of fusion protein was 21.40%, 30.60%, 35.40%, 35.43% respectively. The fusion protein exhibited a band of 78.6 kD on SDS-PAGE analysis, but was not found in controls.The target protein was mainly existed in the form of inclusion body. CONCLUSION:Rice PAL gene expressing E. coli was established by gentic engineering technique.     

Adhesion of bone marrow stromal cell in children with acute lymphoblastic leukemia

AIM and METHOD: The adhesion behavior of bone marrow stromal cells(BMSC) in children with acute lymphoblastic leukemia(ALL) to bone marrow mononuclear cells(BMMC) and Raji cells were investigated by MTT method. The expression of adhesion molecules ICAM-1 and VCAM-1 were detected by flow cytometry. RESULTS: The adhesion ability of BMSC in acute period of ALL to BMMC was lower than that of control group. The adhesion ability of BMSC in long term remission period of ALL to Raji cells higher than that of control group. The expression of ICAM-1 on the surface of BMSC in acute period of ALL is much lower than that of control group. CONCLUSIONS: The adhesion of BMSC to BMMC or tumor cells in children with ALL was abnormal. The abnormal adhesion behavior might be partially due to the changed expression of adhesion molecules on BMSC.     

Interleukin-1β inhibits AKT to down-regulate eNOS Ser1177 phosphorylation in human umbilical vein endothelial cells

AIM To investigate whether interleukin-1β （IL-1β） regulates endothelial nitric oxide synthase （eNOS） phosphorylation at Ser1177 site in human umbilical vein endothelial cells （HUVECs）, and to explore its possible mechanism. METHODS The HUVECs were randomly divided into normal control group, tumor necrosis factor-α （TNF-α） group, IL-1β group, IL-6 group, SC79 ［protein kinase B （PKB/AKT） specific agonist］ group and SC79+IL-1β group. Western blot was used to determine the protein levels of eNOS, p-eNOS-Ser1177, AKT and p-AKT-Ser473 in the HUVECs. Chemical colorimetry was used to detect the nitric oxide （NO） content in the culture medium of HUVECs. RESULTS No statistically significant difference of p-eNOS-Ser1177 level in HUVECs treated with TNF-α and IL-6 was observed as compared with normal control group （P>0.05）, while the protein level of p-eNOS-Ser1177 in the HUVECs and the content of NO in the culture medium of HUVECs decreased significantly in IL-1β group （P<0.05）, and the protein level of p-AKT-Ser473 in the HUVECs was decreased as compared with normal control group （P<0.05）. The AKT agonist SC79 blocked the down-regulation effect of IL-1β on p-eNOS-Ser1177 level in the HUVECs and NO content in the culture medium of HUVECs （P<0.05）. CONCLUSION IL-1β down-regulates the protein level of p-eNOS-Ser1177 in HUVECs and affects the activity of eNOS, which may be involved in AKT/eNOS signaling pathway.     

氯丹和灭蚁灵在污染场地中的空间分布研究

氯丹和灭蚁灵是两种杀虫剂类POPs,在我国仍然生产和使用。在多年生产氯丹、灭蚁灵的企业厂区内及周围布设采样点,采取不同深度的土壤进行分析,研究了POPs生产企业土壤中污染物的空间分布规律。结果表明,生产厂区内污染严重,在土壤表层,靠近生产车间的土壤中氯丹的浓度高达2927.95mg·kg-1,灭蚁灵浓度高达61.90mg·kg-1,并且已经扩散到周边土壤。在土壤深层,氯丹和灭蚁灵有明显的垂直扩散,10m深处土壤均能检出两种污染物。由于灭蚁灵产量小于氯丹产量,灭蚁灵的污染不及氯丹的污染严重。     

莫比朗在柑桔和土壤中的残留动态研究

采用田间试验方法,研究了莫比朗在柑桔和土壤中的消解动态和最终残留。样品用乙腈提取,过Envi-Florial固相小柱净化,液相色谱紫外检测器检测,外标法定量。残留动态试验结果表明,施药浓度为推荐剂量的两倍时(有效成分120g·hm-2),莫比朗在柑桔中的半衰期分别为7.00～8.90d,土壤中为4.00～6.38d。在有效成分为60g·hm-2的剂量下,施药3次,施药后第21d柑桔中莫比朗残留量低于0.10mg·kg-1。     

环境基准值的科学研究与我国环境标准的修订

对环境基准和环境标准在概念上进行了辨析,论述了环境基准科学研究中需要把握的几个核心问题,其中最为关键的一步是不同类型生态受体的选取,因此环境基准值应该是基于不同保护对象的多目标函数或一个范围值。鉴于我国环境标准迫切需要修订这一实际,呼吁加强环境基准的研究,尤其是要开展一次国家层面上的土壤环境基准的系统研究。