Adsorption and affinity of Escherichia coli to different aggregate sizes of a silty clay soil

The sediments produced by water erosion are the main source of pollution of agricultural origin of surface water bodies. These sediments may be associated to bacteria, compromising the quality of nearby water bodies. Therefore, to understand this biological contamination, it is necessary to find out the adsorption capacity and bacterial affmity to aggregate sizes that may result in a differential sedimentation. To this end, in the present work, the distribution, adsorption capacity and affinity to different aggregate sizes of two strains of Escherichia coli in two liquid media of contrasting ionic strength were evaluated in a silty clay soil. The 〈2 μm fraction showed a higher proportion of bacteria than the other aggregate sizes （48%）, whereas among the fractions 〉2 μm, the 20-50μm fraction was the one that showed the highest bacterial adsorption in both liquid media （37.9%）. On the other hand, the highest values of bacterial affinity were found in the 20 to 50 lain fraction （coarse silt） in the low ionic strength media and 20--50 and 〉50 μm in the high ionic strength media. However, the bacterial strains used revealed only some trends in the modification of these variables. This work contributes to the development and implementation of strategies to mitigate pollution, such as control of sediment generation and its subsequent capture in filter strips.     

太平洋牡蛎中大肠杆菌净化的实验研究

静水中人工感染太平洋牡蛎,使其体内埃希氏大肠杆菌(Escherichia coli)浓度达到105 MPN/100g,然后放入紫外线循环系统中对其进行净化,研究贝水质量比(1∶3,1∶6,1∶10,1∶50)、水交换频率(1,3,5次/h)、温度(10,15,20,25℃)及净化层数(上层,中层,下层)等不同环境因子对牡蛎体内大肠杆菌净化的影响。在前6h大肠杆菌净化迅速,之后速度减慢,至36h时能净化2～3个对数值。不同环境因子的净化实验结果表明,当贝水比为1∶6,水交换律为3次/h,温度为15或25℃,净化层数为上层时净化效果最佳。     

Enhanced resistance of marine shrimp Exopalamon carincauda Holthuis to WSSV by injecting live VP28-recombinant bacteria

Recombinant bacteria secreting the transmembrane-truncated region of white spot syndrome virus (WSSV) envelope protein VP28 named DhpVB were constructed, using live Escherichia coli as a deliverer and a constitutive secretory expression plasmid as the expression vector. With the ability to deliver recombinant protein products outside, DhpVB could make VP28 directly interact with the shrimp when injected into shrimp. In order to test the protection potential, live DhpVB were injected into the marine shrimp Exopalamon carincauda Holthuis for three times and WSSV challenge was performed twice simultaneously at the last two injections of the bacteria. The results showed that DhpVB displayed statistically significant advantage over bacteria without VP28 after the second challenge with an average RI (Resistance index) of 0.67 ± 0.08, compared with 0.41 ± 0.09 of bacteria without VP28 (P <0.05). The data suggested that a quasi-immune response may exist in E. carincauda and live bacteria carrying VP28 could enhance the shrimp resistance against WSSV.     

成都府河水体细菌动态变化及耐药性

将府河从都江堰宝瓶口段至望江公园段分设9个监测点,在秋冬、春夏两季分别取水,首次建立由细菌总数、总大肠菌群数及粪大肠菌群数三个微生物学指标组成的体系,对府河两季水质进行对比监测分析,以综合评价水质、分析季节影响因素并反映出污染物来源.以都江堰宝瓶口大肠杆菌耐药率作为背景值,对整个河段的9个采样点分离纯化鉴定得到102株大肠杆菌,用K-B法进行对比药敏试验.结果表明,府河从都江堰段起,穿过成都市区后,至望江公园段细菌总数、大肠菌群数及粪大肠菌群数虽有波动,但细菌总数总体呈增加趋势,春夏季府河营养富集,水质有所降低.府河中大肠杆菌耐药性较小,β-内酰胺类耐药率在0.96％～33.33％之间,氨基糖苷类1.39％～7.84％,氟喹诺酮类11.43％～11.76％,磺胺类复方新诺明为52.78％,其中β-内酰胺类耐药性、氟喹诺酮类耐药性与源头水质有关,其余与医疗机构排放的废水及生活污水有关,此研究为控制成都府河水体中的污染防控提供了理论依据.     

中国对虾素在大肠杆菌(Escherichia coli)中的重组表达

中国对虾素是从中国对虾血细胞中克隆得到的一种抗菌肽。为了进一步研究中国对虾素(CHP)的功能并为制备特异性抗体作准备，采用大肠杆菌表达外源蛋白的方法，进行了对虾素原核表达的研究。根据从中国对虾血细胞中克隆得到的对虾素基因设计特异性引物扩增中国对虾素成熟肽基因(CHP)，插入原核表达载体pGEX-4T-1中，在E．coli BL21表达融合蛋白GST-CHP．结果表明，不同表达菌株的融合蛋白表达量为30％—34％，同一菌株在诱导5h后能达到最高表达量。利用GST亲和层析纯化融合蛋白，将融合蛋白用凝血酶裂解以得到CHP，其分子量约为5．6kDa，N-端测序结果与期望的成熟对虾肽序列一致。用液体生长抑制方法检测活性，表明重组GST-CHP蛋白及CHP均表现出对大肠杆菌的抑菌活性。     

杜氏藻与大肠杆菌跨界融合初探

以链霉蛋白酶制得了杜氏藻(嗜盐单胞绿藻)的原生质体。采用聚乙二醇(PEG)-钙离子法将它们与大肠杆菌(含氯霉素乙酰转移酶基因质粒)的球1,20,1状体进行融合。“融合子”酷似杜氏藻。它们对氯霉素抗性的显著提高寓示了大肠杆菌的质粒已转移到“融合子”细胞中;而且,“融合子”的蛋白质组分与杜氏藻和大肠杆菌相比已发生了较明显的变化。     

具荧光活性的节旋藻藻蓝蛋白α亚基在大肠杆菌中的重组表达

为探索节旋藻藻蓝蛋白的生物合成机理,以钝顶节旋藻(Arthrospira platensisFACHB314)基因组DNA为模板克隆了藻蓝蛋白α亚基基因cpcA、催化脱辅基蛋白与色基结合的裂合酶基因cpcE和cpcF,以及催化色基合成的铁氧蛋白氧化还原酶基因pcyA;以Synechocystissp.PCC6803基因组DNA为模板克隆了亚铁血红素氧化酶基因hox1。然后分别将cpcA、cpcE和cpcF基因构建到质粒pACYCDuet-1中,将hox1和pcyA基因构建到质粒pET-24a(+)中,用电击法将二者共同转化E.coliBL21(DE3),经诱导表达得到具有荧光活性的节旋藻藻蓝蛋白α亚基。在590 nm激发波长下,荧光发射峰为637.8 nm,证实了节旋藻自身的裂合酶CpcE和CpcF能够催化色基PCB与藻蓝蛋白脱辅基结合产生有荧光活性的藻蓝蛋白α亚基,为表达有荧光活性的藻蓝蛋白提供理论和实验基础。     

间接酶联免疫吸附技术检测活的非 可培养状态的大肠杆菌O157：H7

大肠杆菌O157H7在低温贫营养的条件下可进 入活的非可培养状态（Viable but nonculturable state,VBNC），用常规的培养法无法将其检出。作者利用间接酶联免疫吸附技术（Indirect Enzyme-Linked Immunosorbent Assa y, ELISA）检测VBNC的大肠杆菌O157H7，发现此方法可以快速有效地将其检出，最小检测 浓度为10     

沼泽红假单胞菌(Rhodopseudomonas palustris)核酮糖-1,5-二磷酸羧化酶/氧化酶基因克隆及其在大肠杆菌中的表达

核酮糖-1,5-二磷酸羧化酶/氧化酶(RubisCO)(EC4.1.139)是光合细菌通过卡尔文循环固定二氧化碳的关键酶。本文采用PCR方法,从沼泽红假单胞菌株No.9中克隆到RubisCO基因(cbbM)序列(该序列已提交GenBank,登录号:GU061327)。采用同源建模法,建立了该RubisCO蛋白的三维结构模型,预测其活性位点。将cbbM基因亚克隆到表达载体pTV118N上,构建表达质粒pTV-CBBM,转化大肠杆菌BL21(DE3),获得表达菌株BL21(DE3)/pTV-CBBM,该菌株经IPTG诱导表达后,进行SDS-PAGE检测。采用气相色谱法测定破菌上清中的RubisCO酶活。结果表明:①cbbM基因编码461个氨基酸,与沼泽红假单胞菌株DCP3和DH1的RubisCO蛋白序列相似性分别为98%和99%;②推测沼泽红假单胞菌No.9中RubisCO蛋白的活性中心由Asn112、Lys192、Asp194、Glu195、His288、Arg289、His322、Gly424、Ser369、Gly370和Gly394等氨基酸残基组成;③重组蛋白分子量约为50kDa左右,与预测相符;④破菌上清中的RubisCO酶比活高于原始菌株中的酶活,说明目的基因在大肠杆菌中得到了有效表达。     

海南省三亚湾和榆林湾海水中叶绿素α浓度总细菌和大肠杆菌的丰度与 …

１９９６年８月在海南省三亚湾和榆林湾对海水中的叶绿素α浓度,总细菌和大肠杆菌的丰度进行了检测。结果表明叶绿素α浓度和总细菌工以三亚河口和榆林湾内港为最高,离岸逐渐降低。三亚河口和榆林湾内港的叶绿素α高浓度,高的总细菌丰度和三亚河口高的大肠杆菌丰度表明其水体已呈现富营养化,这主要是由生活污水通过三亚河对河口和内港的造成的。