节杆菌PJ3基因组中发现2,3-二羟基联苯双加氧酶基因

从污水中分离出一株以咔唑为惟一碳源和氮源生长的菌株PJ3, 用 16S rDNA 鉴定为节杆菌(Arthrobacter sp), 构建了PJ3菌株基因组文库并获得一株阳性克隆JM109 (pUCW402). 通过GenSCAN软件和BLAST分析发现, 在插入pUCW402的外源片段(3360 bp)中存在一个由933 bp片段编码的2,3-二羟基联苯双加氧酶基因. 进化分析表明, 从PJ3来源的2,3-二羟基联苯双加氧酶在进化树上形成一个独立的分支. Southern杂交进一步证实, 编码该酶的基因定位于节杆菌PJ3的基因组DNA上. 为了鉴定该基因的生物学功能, 构建了BL21 (pETW-8)重组菌株. 与菌株JM109 (pUCW402)和PJ3相比, 2,3-二羟基联苯双加氧酶的表达水平在BL21 (pETW-8)中最高. 酶活分析表明, 2,3-二羟基联苯双加氧酶不具有绝对专一性, 对2,3-二羟基联苯的催化能力高于邻苯二酚. 因此推测PJ3菌株对芳香族化合物降解过程中, 2,3-二羟基联苯双加氧酶主要负责催化双环化合物的降解.     

Eisenia fetida中蚯蚓纤溶酶P-Ⅲ组分序列克隆与分析

从Eisenia fetida中提取总RNA,利用P-Ⅲ组分序列两端的保守性设计引物,通过RT-PCR获得P-Ⅲ组分基因,并构建该组分质粒(pUC18)文库,经PCR筛选后进行序列测定及分析。在原已获得的Efp-Ⅲ-1和Efp-Ⅲ-3的基础上,利用中间引物从文库中筛选到Efp-Ⅲ-2基因。序列分析表明,该基因编码序列为720bp,编码239个氨基酸,与GenBank报道的Efp-Ⅲ-2五条序列整体相似性高达99.02%。通过序列比对,发现Efp-Ⅲ-1、2和3具有完全相同的结构域,差异位点零星分布在序列中间本文首次由一个实验室获得P-Ⅲ组分的三个亚组序列,序列比对分析证明它们是蚯蚓体内P-Ⅲ组分基因多态性的表现。     

濒危植物桫椤过氧化物酶的分离纯化与性质

将齿叶黑桫椤叶片提取液经硫酸铵盐析、透析、DEAE-琼脂糖凝胶柱层析和Sephacryl S-200凝胶柱层析分离得到纯化的POD,其纯化倍数为26.29,回收率为27.04%;通过SDS-PAGE法测得POD的分子量为43000D;通过IEF-PAGE法测得POD的等电点为4.55;齿叶黑桫椤POD最适pH在4左右;最适温度在45℃左右;Km值在最适温度45℃、最适pH4的条件下为0.07 mol/L。     

不同温度对双齿多刺蚁养殖及其保护酶系的影响

在不同温度下长期养殖双齿多刺蚁,观察其存活率和活动状况,测定了不同温度对其SOD,POD,CAT活力的影响,并进行了保护酶活性和存活率的相关分析.实验表明,在较低和较高温度下其存活率明显低于10-40℃测定值.在40-45℃时,酶的活性随着温度升高急速下降;在-5-10℃时,酶的活性随着温度降低而下降.保护酶活性和存活率密切相关.10℃以下的低温和40℃以上的高温对其有不同程度的伤害,其适宜温区为10-40℃,最适温度为20-30℃.     

Icariin suppresses bone resorption activity of rabbit osteoclasts in vitro

The effect of icariin on the bone resorption activity of rabbit osteoclasts is assessed in vitro. Osteoclasts were isolated from Japanese white rabbits and cultured on plates with a sterilized bone slice in each well. After treatment with icariin at various concentrations, the bone resorption activity of osteoclasts was evaluated by examining pit areas, superoxide anion (·O2-) generation, size and number of actin rings and intracellular calcium concentration [Ca2 ]i. As revealed by these data, icariin elicited continuous decline of [Ca2 ]i, making actin ring constricted and ·O2- generation decreased. These events resulted in smaller and fewer pits which indicate suppressed bone resorption activity of rabbit osteoclasts by icariin.     

2,3-Dihydroxybiphenyl dioxygenase gene was first discovered in Arthrobacter sp. strain PJ3

Bacterium strain PJ3,isolated from wastewater and identified as Arthrobacter sp. bacterium based on its 16S rDNA gene,could use carbazole as the sole carbon,nitrogen and energy source. The genomic library of strain PJ3 was constructed and a positive clone JM109(pUCW402) was screened out for the expression of dioxygenase by the ability to form yellow ring-fission product. A 2,3-dihydroxybiphenyl dioxygenase(23DHBD) gene of 933 bp was found in the 3360 bp exogenous fragment of pUCW402 by GenSCAN software and BLAST analysis. The phylogenetic analysis showed that 23DHBD from strain PJ3 formed a deep branch separate from a cluster containing most known 23DHBD in GenBank. Southern hybridization confirmed for the first time that the 23DHBD gene was from the genomic DNA of Arthrobacter sp. PJ3. In order to test the gene function,recombinant bacterium BL21(pETW-8) was constructed to express 23DHBD. The expression level in BL21(pETW-8) was highest compared with the recombinant bacteria JM109(pUCW402) and strain PJ3. We observed that 23DHBD was not absolute specific. The enzyme activity was higher with 2,3-dihydroxybiphenyl as a substrate than with catechol. The substrate specificity assay suggested that 23DHBD was essential for cleavage of bi-cyclic aromatic compounds during the course of aromatic compound biodegradation in Arthrobacter sp. strain PJ3.     

动物源性食品的氯霉素ELISA检测及分析

采用酶联免疫法检测了730例天津口岸进出口肉制品、水产品、乳制品和蜂蜜等几种动物源性食品中的氯霉素含量,对阳性结果经液相色谱-质谱联用EEC Cy3.6 confirming method进行确证,分析了其残留现状,并对英国RANDOX公司氯霉素检测试剂盒从检测范围、B/B050%抑制浓度、板内变异、板间变异、样品添加试验、实际样品检测、稳定性等方面各项指标进行试验.结果表明,试剂盒的检测低限为0.1 ng/mL,线性范围为0.05～4.7 ng/mL,B/B050%抑制浓度为0.4 ng/mL,样品加标回收率为80.5%～110%,板内变异系数小于5%,板间变异系数小于10%,试剂盒稳定性好.     

牛奶中氨基苷柱后衍生高效液相色谱法的建立

建立了庆大霉素（氨基苷类抗生素）在牛奶中残留量柱后衍生高效液相色谱检测法。牛奶样品采用磷酸缓冲液提取，三氯乙酸沉淀蛋白，固相萃取净化，甲醇洗脱氮气吹干。反相高效液相色谱法分离柱后衍生荧光检测。该方法简化了前处理步骤，缩短了分析时间，并促进了在线自动化分析技术的开发。本方法的线性、准确性和精密度以及检测限和定量限均符合标准方法的要求，适用于牛奶中庆大霉素残留量的检测。     

超声波刺激对绞股蓝幼苗的POD同工酶影响

运用聚丙烯酰胺凝胶电泳(PAGE)方法研究了用不同时间的超声波处理对绞股蓝幼苗的过氧化物酶(POD)同工酶表达的影响.结果表明,适当剂量的超声波处理能够促进绞股蓝(Gynostemma pentaphyllum)过氧化物酶同工酶的表达;超声波处理后的绞股蓝与对照相比,过氧化物酶同工酶在酶带数目和活性上都有明显增加,尤其以T6和T12的条带数最多且染色较深.     

茜草提取物对训练大鼠不同组织NO含量及血清酶活性的影响

通过建立力竭跑台训练模型,测定大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、乳酸脱氢酶(LDH)、肌酸激酶(CK)活性和不同组织中NO含量,研究了茜草提取物对大强度耐力训练大鼠血清酶活性和运动能力的影响.结果显示,与安静组比较,运动力竭组大鼠血清ALT、AST、LDH和CK活性显著升高(P<0.01),不同组织NO含量较显著升高(P<0.05);与训练力竭组比较,茜草提取物组大鼠血清ALT、AST和CK活性有较显著下降(P<0.05),不同组织NO含量较显著升高(P<0.05),使大鼠运动能力有了明显提高.