基于蛋白质反式剪接在大肠杆菌中表达鲎C因子CES多肽

鲎C因子是鲎血细胞中一种对内毒素具有高亲和力的丝氨酸蛋白酶原,可替代鲎试剂用于内毒素检测。鲎C因子N端Cys—EGF．sushil—sushi2（CES）片段中的sushil区域是与内毒素结合的关键部位。本研究在sushil结构域中的特定位点断开CES与内毒素结合的功能片段,然后分别和蛋白质内含肽SspDnaX的N端片段（IN）和C端片段0C）连接,并在大肠杆菌中表达,表达产物经亲和层析纯化、复性以及内毒素去除后,利用蛋白质反式剪接系统,在体外诱导该蛋白质内含肽发生剪接,重新得到C因子的CES蛋白,并检测其活性。实验结果表明此重组CES蛋白具有结合内毒素活性,提示在大肠杆菌系统中开发低成本内毒素检测试剂的可行性。     

蚯蚓与黑麦草相互作用对土壤中荧蒽去除的影响

Earthworms can promote the bioremediation of contaminated soils through enhancing plant growth and microorganism development. The individual and combined effects of earthworms and ryegrass (Lolium multiflorum Lam.) on the removal of fluoranthene from a sandy-loam alluvial soil were investigated in a 70-d microcosm experiment. The experiment was set up in a complete factorial design with treatments in four replicates: without earthworms or ryegrass (control, CK), with earthworms only (E), with ryegrass only (P), and with both earthworms and ryegrass (EP). The residual fluoranthene, microbial biomass C, and polyphenol oxidase activity in the soil changed significantly (P<0.01) with time. In general, the residual concentration of fluoranthene in the soil decreased sharply from 71.8-88.7 to 31.7-37.4 mg kg 1 in 14 d, and then decreased gradually to 19.7-30.5 mg kg 1 on the 70th d. The fluoranthene concentration left in the soil was the least with both earthworms and ryegrass, compared to the other treatments at the end of the experiment. Half-life times of fluoranthene in the E, P, and EP treatments were 17.8%-36.3% smaller than that of CK. More fluoranthene was absorbed by earthworms than ryegrass. However, the total amounts of fluoranthene accumulated in both the ryegrass and earthworms were small, only accounting for 0.01%-1.20% of the lost fluoranthene. Therefore, we assumed that microbial degradation would play a dominant functional role in fluoranthene removal from soil. We found that earthworms significantly increased microbial biomass C and polyphenol oxidase activity (P<0.01) in the presence of ryegrass at the end of the experiment. Furthermore, microbial biomass C and polyphenol oxidase activity were significantly (P<0.05) and negatively related to the residual fluoranthene concentration. This implied that earthworms might promote the removal of fluoranthene from soil via stimulating microbial biomass C and polyphenol oxidase activity.     

华北地区春玉米–冬绿肥轮作对碳、氮蓄积和土壤养分以及微生物的影响

华北农田纳入冬绿肥能够提升土壤质量和抑制土壤风蚀。本文主要以一年一茬春玉米为例,将毛苕子（Vicia villosa Roth）与二月兰（诸葛菜,Oryehophragmus violaeeus）作为冬绿肥,研究春玉米冬绿肥轮作模式下的碳氮蓄积量及土壤养分和微生物的变化。2 年试验结果表明,冬绿肥区碳蓄积量是冬闲区的2.4～3.6倍;氮蓄积量约为冬闲区的4.0～4.7倍;二月兰区碳蓄积量比毛苕子区高32.7%～39.2%,毛苕子区氮蓄积量比二月兰区高约5.7%,差异显著（碳蓄积量是指在该轮作系统下植物体的碳素累积量;氮蓄积量是指在该轮作系统下植物体的氮素累积量）。春玉米种植制度短期纳入冬绿肥作物对玉米产量及对土壤pH,速效氮、速效磷含量、土壤总碳量无显著影响。冬绿肥区有机碳含量比冬闲区提高4.5%～5.7%,且二月兰区毛苕子区,差异显著;冬绿肥显著提高了土壤微生物数量,以细菌数量变化最为明显。冬绿肥区细菌数量是冬闲区的2.0～3.7倍,毛苕子区二月兰区。说明农田短期纳入冬绿肥,可以提高土壤有机碳含量,促进土壤微生物的活动,有利于保持土壤肥力。     

融合蛋白C3d提高新城疫灭活苗的免疫原性

补体C3d是补体系统（complement system,CS）中补体c3的最终裂解产物。本试验利用RT—PCR技术从从肉鸡肝脏总RNA中扩增C3dcDNA,测序证实为C3d基因。然后将其连接至表达载体pET-32a（＋）,转化E．coliBL21（DE3）,1％IPTG诱导表达。SDS-PAGE、Western-blotting证实目的蛋白为C3d。亲和层析分离C3d融合蛋白,与纯化新城疫病毒（Lasota株）经戊二醛连接后免疫SPF鸡。血凝抑制试验（HI）检测新城疫抗体水平,MTT法检测胸腺T淋巴细胞转化率。结果表明C3d融合蛋白能够提高特异性抗体水平以及细胞免疫水平,本研究为进一步研究、利用鸡补体C3d奠定了基础。     

可视化专业组件在C++Builder6·0系统开发中的应用

以C++Builder6.0为工具,结合特定行业的实际需求,研究如何开发基于C++Builder6.0的可视化专业组件,并对其界面修饰、艺术造型与动画显示等方面进行研究与分析。     

基于4C营销理论的定制式个人数字图书馆构建原则

随着个人数字图书馆构建实践的发展,在当前信息环境中自建式个人数字图书馆面临的困难越发凸现.为扩大个人数字图书馆的服务范围,充分发挥其个性化信息服务的优势,本文在借鉴4C营销理论的基础上提出定制式个人数字图书馆的构建原则.     

香葱冻干过程中影响因素的研究

对新鲜香葱不同的真空冷冻干燥条件进行了对比实验研究,得到香葱真空冷冻干燥的最佳工艺条件：升华干燥温度90℃、解析温度40℃、压强50Pa。     

硫丹降解菌C7在土壤中的降解性能及其酶促降解试验

Ochrobactrum sp. C7是一株高效硫丹降解菌。本试验对C7在土壤中降解硫丹的效果以及C7产生的硫丹水解酶(粗酶液)的性质进行了研究。结果表明,该菌在30天内能很好地降解土壤中的硫丹;降解酶在30~45℃、pH5.5~7.5的条件下活性较高;在pH7和40℃时显示最大的硫丹水解酶活性。初步判断水解酶的表达可能是组成型的,此粗酶液可用于果蔬表面农药残留的快速解毒和清除,从而为高效降解菌株的应用与环境中的生物修复提供了理论依据。     

灌水对不同穗型小麦碳氮代谢及籽粒灌浆的影响

为了给小麦节水栽培实践提供参考,在大田条件下研究了灌水次数对大穗型小麦品种豫麦66与多穗型小麦品种豫麦49碳氮代谢及籽粒灌浆的影响。结果表明,增加灌水次数可以提高旗叶硝酸还原酶(NR)活性和可溶性蛋白质含量,在灌浆中后期豫麦66较豫麦49具有明显优势。随着灌水次数增加,旗叶和籽粒可溶性糖含量在灌浆前期降低,而在灌浆后期籽粒可溶性糖含量升高,旗叶则表现相反。籽粒淀粉积累速率在灌浆前期随灌水次数增加而降低,但在灌浆后期则与之相反。籽粒灌浆速率呈单峰曲线变化,增加灌水次数,积累高峰期推迟,且前期慢,后期快,在花后35 d豫麦66在灌一水、灌两水和灌三水处理下籽粒灌浆速率分别为0.96、1.24和1.07 mg/d,而豫麦49分别为0.94、0.39和0.36 mg/d。豫麦66与豫麦49号相比,旗叶可溶性糖含量在灌浆中后期较低,淀粉及籽粒积累速率的峰值出现较晚但持续时间较长,表明大穗型品种同化物外运能力和库活性较强,灌浆进程稳健。     

Fine Mapping of C(Chromogen for Anthocyanin)Gene in Rice

Seven residual heterozygous lines(RHLs)displaying different genotypic compositions in the genomic region covering probable locations of C (Chromogen for anthocyanin)gene on the short arm of rice chromosome 6 were selected from the progenies of the indica cross Zhenshan 97B/Milyang 46.Seeds were harvested from each of the seven plants,and the resultant F2:3 populations were used for fine mapping of C gene.It was shown in the populations that the apiculus coloration matched to basal leaf sheath coloration in each plant.By relating the coloration performances of the populations with the genotypic compositions of the RHLS,the C locus was located between rice SSR markers RM314 and RM253.By using a total of 1279 F2:3 individuals from two populations showing coloration segregation.the C locus was then located between RM111 and RM253,with genetic distances of 0.7 cM to RM111 and 0.4 cM to RM253.Twenty-two recombinants found in the two populations were assayed with seven more markers located between RM111 and RM253,including six SSR markers and one marker for the C gene candidate,OsC1.The C locus Was delimited to a 59.3-kb region in which OsC1 was located.