猪链球菌蛋白表面展示系统的构建及展示蛋白的初步观察

为构建猪链球菌(Streptococcus suis)蛋白表面展示系统,本研究通过序列分析,确定猪链球菌的LPxTG蛋白及其信号肽(SP)和胞壁锚定基序(CWA),通过PCR扩增Peno-SP、GFP、CWA的DNA片段并融合,构建强启动子Peno控制表达编码SP-GFP-CWA融合蛋白的DNA片段,将该重组DNA片段连接pSET2载体,获得蛋白表面展示质粒,转化猪链球菌,构建得到以GFP为报告蛋白的猪链球菌蛋白表面展示系统。结果显示,利用猪链球菌的10个LPxTG蛋白及其SP和CWA序列,构建了10个含有Peno-SP-GFP-CWA融合片段的重组pSET2表面蛋白展示质粒pSsPSD1至pSsPSD10,分别转化猪链球菌05ZYH33,PCR鉴定显示其中7个转化猪链球菌。采用western blot初步检测其展示蛋白,结果显示,7个转化阳性菌株均能有效表达GFP蛋白,以成熟GFP条带为指标,均表现出了一定的外源GFP表面展示水平,分别命名为SsPSD1、SsPSD2、SsPSD4、SsPSD7-SsPSD10,其中SsPSD1、SsPSD4、SsPSD8和SsPSD9表面展示水平相对较好,在猪链球菌表面展示外源蛋白方面具有很好的潜力。本研究首次尝试建立猪链球菌蛋白表面展示系统,为猪链球菌表面递呈外源蛋白或抗原提供了新的策略。     

Rapid mapping of candidate genes for cold tolerance in Oryza rufipogon Griff. by QTL-seq of seedlings

Cold stress is a major problem in rice production. To rapidly identify genes for cold tolerance in Dongxiang wild rice(DWR, Oryza rufipogon Griff.), sequencing-based bulked segregant analysis of QTL-seq method was used to resequence the extremely resistant(R) and susceptible(S) bulks of a backcross inbred lines(BILs) population(derived from Oryza sativa×O. rufipogon) and their parents. Single nucleotide polymorphisms(SNP)-index graphs and corresponding Δ(SNPindex) graphs(at 99 and 95% confidence levels) for R-and S-bulks detected a total of 2 609 candidate SNPs, including 58 candidate cold-tolerance genes. Quantitative real-time PCR analysis revealed that 5 out of the 58 candidate genes had significant differences in expression between O. sativa and O. rufipogon. Structural variation and functional annotations of the 5 candidate genes were also analyzed, and allowed us to identify 2 insertion-deletion(InDel) markers(12-7 and 12-16) that were linked with candidate genes on chromosome 12 in DWR. These results are helpful for cloning and using cold tolerance genes from common wild rice in cultivated rice.     

Effect of Dietary Lactobacillus fermentum from Pig on Growth Performance,Nutrient Apparent Digestibility,Faecal Microflora Number and Serum Immune Indices in Weaned Piglets

This experiment was conducted to study the effects of Lactobacillus fermentum on growth performance, nutrient apparent digestibility, faecal microflora number and serum immune indices of weaned piglets.A total of 160 weaned piglets with an average age of (30±2) days and averge body weight of (7.85±0.68) kg were randomly distributed into 4 groups with 4 replicates per group and 10 piglets per replicate feeding basal diet, 0.1%, 0.2%, and 0.3% Lactobacillus fermentum supplementation diets, respectively.The experiment lasted for 35 d.The results showed that, compared with the control group, the average daily gain, the feed conversion ratio, the apparent digestibility of crude protein, calcium and total phosphorusin of Lactobacillus fermentum groups were all increased in varying degrees.The optimum supplemental level of Lactobacillus fermentum should be 0.2%, and at this level, the average daily gain was significantly increased by 8.70% (P<0.05), the F/G was decreased by 4.62% (P<0.05), the apparent digestibility of crude protein, calcium and total phosphorus were significantly increased (P<0.05), the number of faecal E.coli was significantly decreased (P<0.05), and the serum IgG was significantly increased by 8.39% (P<0.05).The results indicated that Lactobacillus fermentum supplementation could improve the immune function, growth performance, the apparent digestibility of crude protein, calcium and total phosphorus and reduce the number of faecal E.coli of weaned piglets.     

小麦面粉中赭曲霉毒素A的CdSe/ZnS荧光量子点免疫探针检测

构建一种基于荧光量子点免疫探针的小麦面粉中赭曲霉毒素A(OTA)的检测方法。以EDC为活化剂将CdSe/ZnS半导体荧光量子点与OTA单克隆抗体偶联,制备OTA检测探针并评价其检测性能。结果表明:制备的检测探针在3d保存期内具有良好的稳定性。当OTA质量浓度为0.1~0.9μg/L时,检测探针的荧光强度与OTA质量浓度呈线性关系,其相关系数为0.996 3,检出限为0.017 7μg/L。与国标法对照的结果说明,此方法具有可行性。小麦面粉中的加标回收试验结果显示,OTA的回收率为96.0%~102.7%,相对标准偏差不超过4.1%,说明准确性良好。可见,OTA的CdSe/ZnS荧光量子点免疫探针检测方法操作简单、准确度高、结果稳定,有望应用于小麦面粉中OTA的检测。     

保定地区几个早熟苹果品种(系)生长观察

为筛选适宜保定地区栽植的早熟苹果品种,以嫁接在SH40中间砧上的不同早熟苹果品种(系)为试材,测定了不同品种(系)早熟苹果树体生长发育指标和果实品质。结果表明:除嘎啦早熟变异外,其他供试品种(系)树体生长较好,3年生时Xu2-7、Xu3-2树高330.0~352.5 cm,4年生时为394.0~417.5 cm,Xu3-2、藤牧1号、信浓红4年生为336.5~381.0 cm。4年生时,干径以Xu2-5最大;短枝、中枝、叶丛枝均以Xu2-5最多,长枝以Xu2-7最多,总枝量以Xu2-5最多;藤牧1号短枝所占比例最高(57.4%)。应用模糊综合评判法得出Xu2-5、Xu2-7、信浓红、藤牧1号果实品质较高。     

养殖废弃物生物有机肥料对藜蒿产量及肥料利用率的影响

研究了养殖废弃物生物有机肥料和化肥配施与单施化肥对云南藜蒿产量、相对干物质含量、土壤养分含量及肥料利用率的影响。试验结果表明,各施肥处理的藜蒿产量(21 273~29 110 kg/hm~2)均显著高于对照(17 765 kg/hm~2),其中有机无机肥配施处理的藜蒿产量(20 334~29 110 kg/hm~2)比单施化肥处理(21 273 kg/hm~2)增加19.8%~36.8%;同时,有机无机肥配施处理的藜蒿相对干物质含量比单施化肥处理提高了1.14%~5.48%;与单施化肥处理相比,有机无机肥配施处理明显提高了土壤养分含量、改善了土壤供肥特性、提高了肥料利用率;藜蒿地上部的养分积累量、肥料回收率和肥料农学利用效率及生理利用效率均显著提高。     

Whole-genome identiifcation and expression analysis of K+eflfux antiporter (KEA) and Na+/H+antiporter (NHX) families under abiotic stress in soybean

Sodium toxicity and potassium insufifcient are important factors affecting the growth and development of soybean in saline soil. As the capacity of plants to maintain a high cytosolic, K+/Na+ratio is t...     

Effect of AIR on Inflammatory Reaction Infected by Brucella

In order to investigate the effect of AIR on inflammatory reaction infected by Brucellamelitensis (16M), the AIR domain of Tecpr1 gene of murine macrophages RAW264.7 were knocked down (I-A), overexpressed (O-A) and reversed (OA-IA). Using the chlorine fluorescein (DCFH-DA) as a probe, we detected the variation of ROS production and mitochondria distribution by confocal laser scanning microscopy. We observed the expression changes of NLRP3, ASC and Caspase-1 by qRT-PCR and the expression changes of IL-18,IL-1β and Caspase-1 in host cells by ELISA. The results showed that 16M could stimulate RAW264.7 cells to produce ROS by time-dependent pathway, and I-A group and O-A group showed more abnormal accumulation of mitochondrial. The results of qRT-PCR and ELISA suggested that it had effect on the expression levels of NLRP3, ASC,Caspase-1 and IL-18, IL-1β and Caspase-1 in cells of different groups. Those results indicated that with AIR gene deletion, the release amount of ROS changed, mitochondrial clustered abnormally, and AIR was closely related to the activation of inflammasomes and induction of inflammatory reactions.     

表达鸡α干扰素重组酿酒酵母菌的构建与鉴定

通过构建鸡α干扰素真核表达载体,以获得能够表达鸡α干扰素的酵母工程菌株。首先根据GenBank公布的鸡α干扰素基因序列设计上、下游引物,以略阳乌鸡基因组DNA为模板,PCR扩增鸡α干扰素基因片段。然后,利用基因重组技术将鸡α-干扰素基因插入真核表达载体,通过菌落PCR和DNA测序确定目标载体,构建成功后转化至酿酒酵母AH109,再通过营养缺陷培养、质粒提取和PCR鉴定带JMB440-ChIFN-α载体的重组酵母菌株。最终通过酵母培养、总蛋白收集、SDS-PAGE电泳和Western blot等,确定鸡α干扰素基因能够与酵母菌株重组,并表达α干扰素,蛋白条带大小约为23ku,与预期结果一致,表明成功构建了表达鸡干扰素基因的工程酵母菌株。