表达牛病毒性腹泻病毒E2蛋白的重组乳酸菌的构建及蛋白免疫原性分析     

吴桐忠  努尔赛力克&#  努素甫  黄新  韩猛立  张星星  张倩  王新华  何延华  钟发刚 《中国畜牧兽医》2021,48(8):2975-2981

试验旨在构建能表达牛病毒性腹泻病毒（Bovine viral diarrhea virus，BVDV）E2抗原蛋白的重组乳酸乳球菌（Lactococcus lactis），为进一步研制BVDV乳酸菌口服活载体疫苗奠定基础。将BVDV E2基因克隆后测序，根据乳酸乳球菌的密码子偏嗜性进行优化，再将优化的基因片段插入表达载体pNZ8148中，并电转化乳酸乳球菌NZ9000感受态细胞，构建重组乳酸菌pNZ8148-E2/NZ9000，经1 ng/mL乳链菌肽诱导表达后，对菌体物进行了SDS-PAGE和Western blotting分析。将重组乳酸菌pNZ8148-E2/NZ9000口服免疫6~12月龄健康犊牛，在免疫后不同时间点采集血液样品并分离血清，用间接ELISA方法检测抗体水平。结果显示，PCR扩增到了1 149 bp的目的片段，乳酸菌密码子偏嗜性优化后，GC含量从45.28%变为34.30%。重组质粒pNZ8148-E2经酶切鉴定插入片段与预期大小相符，在菌体裂解物中出现大小约42 ku的条带，与预期蛋白大小一致，且该蛋白可与BVDV E2抗体反应。在免疫犊牛的血清中检测到特异性抗BVDV E2蛋白的抗体。本研究结果表明，表达BVDV E2蛋白的重组乳酸菌口服免疫可诱导犊牛产生特异性的体液免疫反应，该重组菌具有较好的免疫原性。  相似文献   

TCDCA对IL-1β刺激下AA大鼠成纤维样滑膜细胞中PGE2分泌的影响     

TCDCA对IL  刺激下AA大鼠成纤维样滑膜细胞中PGE分泌的影响 《畜牧与饲料科学》2021,42(5):1-5

[目的] 进一步探讨牛磺鹅去氧胆酸（taurochenodeoxycholic acid,TCDCA）在抗炎免疫方面的潜在调节作用。[方法] 以AA大鼠成纤维样滑膜细胞作为研究对象,采用ELISA方法检测TCDCA和IL-1β作用下AA大鼠成纤维样滑膜细胞上清液中PGE₂的含量,分析TCDCA对IL-1β刺激下AA大鼠成纤维样滑膜细胞中PGE₂分泌情况的影响。[结果] TCDCA能够对IL-1β刺激下AA大鼠纤维样滑膜细胞PGE₂的分泌产生下调作用（P<0.05）。[结论] TCDCA对IL-1β刺激下AA大鼠成纤维样滑膜细胞中PGE₂的分泌具有抑制作用,为TCDCA在兽医临床应用提供依据。  相似文献   

牛病毒性腹泻病毒E2蛋白纳米抗体筛选及反应原性检测     

李岩  肖盛中  杨艳  张彦红  蔡卓轩  周子恒  张哲  盛金良 《畜牧与兽医》2021,(3):71-76

通过噬菌体展示技术筛选牛病毒性腹泻病毒(BVDV)重组E2蛋白特异性纳米抗体,验证纳米抗体反应原性。使用BVDV灭活疫苗免疫羊驼,分别在第0、21、49及70天采集全血,测得抗体效价后分离全血中淋巴细胞,提取总RNA,反转录后PCR扩增目的片段。目的片段和pCANTAB5E使用限制性内切酶酶切连接后转至TG1感受态细胞中,应用噬菌体展示技术构建VHH噬菌体展示文库。再经过3轮"吸附-洗脱-筛选"后得到与BVDV-E2结合的噬菌体,用ELISA鉴定其反应性。结果获得插入率为90.8%,库容为1.02×107 CFU/mL的文库。ELISA结果和序列分析显示,得到2条与E2蛋白具有良好反应性的纳米抗体且与VHH同源性较高的序列。研究结果为BVDV的防控和新型疫苗的研制奠定基础。  相似文献   

Dexmedetomidine attenuates acute alcoholic hepatic injury in mice     

HUANG Qiang  XIA Ming-zhu  YANG Wen-jie  DAI Zhong-liang  JIANG Yuan-xu 《园艺学报》2017,33(10):1891-1895

AIM: To investigate the effects of dexmedetomidine (DEX) on acute alcoholic hepatic injury in mice and to explore the possible mechanisms. METHODS: Kunming mice (n=50) were randomly divided into 5 groups (n=10): normal saline control (NS) group, acute alcoholic hepatic injury model (E) group, low-dose (10 μg/kg) DEX (E+L) group, medium-dose (50 μg/kg) DEX (E+M) group and high-dose (100 μg/kg) DEX (E+H) group. The animals were sacrificed at 6 h after gavage of alcohol or normal saline. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) were measured. The livers were removed for evaluation of histological characteristics and determining the content of tumor necrosis factor-α (TNF-α) amd interleukin-1β (IL-1β) in the liver tissues by ELISA. The expression levels of cytochrome P450 2E1 (CYP2E1) and nuclear factor-κB (NF-κB) in the liver tissues were evaluated by Western blot. RESULTS: Compared with NS group, the levels of ALT, AST and TG were obviously increased in E group, which were obviously decreased in E+M and E+H groups. Compared with NS group, the levels of TNF-α, IL-1β and MDA were obviously increase in E group, which were obviously decreased in E+M and E+H groups. Compared with NS group, the activity of SOD and the content of GSH were obviously decreased in E group, which were obviously increased in E+M and E+H groups. Compared with NS group, the expression of CYP2E1 and NF-κB was obviously increase in E group, which was obviously decreased in E+M and E+H groups. Compared with NS group, ethanol induced marked liver histological injury, which was less pronounced in E+M and E+H groups. CONCLUSION: DEX has a protective effect on mouse liver with acute alcoholic injury by the involvement in the processes of antioxidation and antiinflammation, and its mechanism may be associated with the inhibition of CYP2E1 and NF-κB expression.  相似文献   

绿洲灌区不同密度玉米群体的耗水特性研究   总被引：2，自引：0，他引：2  

王巧梅  樊志龙  赵彦华  殷文  柴强 《作物学报》2017,43(9):1347-1356

针对水资源不足严重制约绿洲灌区玉米生产,密植对玉米耗水特性影响研究薄弱,以及生产实践中缺乏调控种植密度以同步提高产量和水分利用效率的理论依据等问题,2012—2015年,以先玉335为参试品种,在相同施肥、灌水制度下,设75 000株hm–2(低,D1)、87 000株hm–2(中,D2)、99 000株hm–2(高,D3)3个密度水平,探讨密度对玉米耗水的时间动态、耗水结构以及利用效率的影响,以期为优化密植增产节水技术提供理论依据。结果表明,随密度增大,玉米全生育期总耗水量(ET)增大,大喇叭口期至吐丝期是耗水量增大的主要时期,高、中密度处理与低密度处理相比,全生育期总耗水量4年平均分别高22.8%、14.4%,大喇叭口期至吐丝期平均分别高28.4%、18.2%,其他生育时期不同处理的耗水量差异不显著。增加密度可降低玉米的无效耗水,提高水分利用率,高、中密度处理较低密度处理全生育期的棵间蒸发量(E)4年平均分别减少56.5 mm、27.6 mm,密植降低棵间蒸发量的主要时期在拔节至灌浆期,其中大喇叭口期至吐丝期的日均棵间蒸发量分别减少0.51 mm、0.27 mm;高、中密度与低密度处理相比,全生育期的E/ET分别降低15.8%、6.2%,其中拔节至大喇叭口期减小幅度最大,分别为22.1%、10.7%。与低密度处理相比,高、中密度处理的籽粒产量分别提高了17.9%、14.8%,但高、中密度处理间的产量差异不显著;高、中、低密度处理的水分利用效率(WUE)分别为18.2、19.3和16.8 kg hm–2 mm–1,中密度处理的WUE显著高于低密度处理;与低密度处理相比,高、中密度处理4年平均灌溉水利用效率(IWUE)分别提高了34.5%、19.6%。本研究说明,在传统供水覆膜条件下,进一步增加种植密度是干旱绿洲灌区提高玉米产量和灌溉水利用效率的可行措施。  相似文献   

The transition time from the juvenile phase to the adult phase differs in soybean cultivars ‘Enrei’ and ‘Peking’          下载免费PDF全文

Suguru Ozawa  Katsunori Hatakeyama  Yoshihito Takahata  Shuji Yokoi 《Plant Breeding》2017,136(3):386-392

Plants develop juvenile phase to adult phase in vegetative stage. Although soybean is a very important crop worldwide, there has been only one study of the juvenile–adult phase change. In this study, we determined that the juvenile–adult phase change occurred at different stages in two soybean cultivars that differ in their photosensitivity. Cultivar ‘Enrei’ (E1e2e3E4) is weakly photosensitive and cultivar ‘Peking’ (E1E2E3E4) is strongly photosensitive. In ‘Enrei’, the leaf size gradually increased at a constant leaf position regardless of the difference in day length. In ‘Peking’ plants transferred to short‐day conditions at several leaf development stages, leaf size gradually increased at different leaf positions. Expression of miR156 by ‘Enrei’ transferred to short‐day conditions had nearly the same pattern as that of ‘Enrei’ grown under long‐day conditions. In ‘Peking’, the expression of miR156 had different patterns in younger leaves of plants subjected to either a short‐day treatment or long‐day conditions. These results indicate that the E2 and E3 loci that regulate photosensitivity also regulate the expression of miR156 and the juvenile–adult phase change in soybean.  相似文献   

欧美杂交杨Pnd-LRR3基因克隆及其抗锈菌侵染表达   总被引：1，自引：0，他引：1  

陈俏丽  李丹蕾  王峰  邹莉  王志英  牛春阳  王博文  零雅茗  崔嵘  张玉洲 《东北林业大学学报》2015,(3):95-98,103

为深入探究欧美杂交杨(Populus nigra×P.deltoides)感病的分子机理,以及抗病基因在发病过程中所起的作用,克隆了欧美杂交杨LRR3蛋白编码基因Pnd-LRR3。对Pnd-LRR3基因进行了生物信息学分析,并对其在抗锈菌过程中的功能进行了研究。Q-PCR结果显示,强致病性的落叶松-杨栅锈菌(Melampsora larici-populina)E4菌株接种6 h,Pnd-LRR3基因表达量上调增幅较大,168 h为显著下调。说明Pnd-LRR3基因在E4侵染过程中起到一定抗性作用,但最终无法阻止锈菌的侵染。  相似文献   

Plant growth regulator and its interactions with environment and genotype affect maize optimal plant density and yield     

《European Journal of Agronomy》2017

Increasing planting density is important to raise maize yield, however, high density often leads to an increase risk of lodging due to dense canopy and weak stem. Maize yield and optimal plant density are increased by applying plant growth regulator compound of ethephon and DA-6, however, we do not know if this compound would interact with location and genotype. In this study, a novel plant growth regulator, as the synthesis of N, N- diethyl − 2 − hexanoyl oxygen radicals − ethyl amine (2-ethyl chloride) phosphonic acid salt (DHEAP), combining the effects of ethephon and DA-6 in one chemical, was developed and tested at three locations, five plant densities (6.75, 8.25, 9.75, 11.25 and 12.75 plants m⁻²) and three cultivars in 2014–2015. This study aimed to quantify the interactions between environment, genotype and management (Appling DHEAP and plant density) on lodging-related optimal plant density and yield. DHEAP significantly increased grain yield by 10.7% due to the increases of kernel weight by 3.2% and kernel number per ear by 4.4%. On average across genotypes and environments, applying DHEAP increased optimum plant density by 6%. The optimal plant density interacted with cultivar, DHEAP and environment. Applying DHEAP reduced lodging percentage by lowering ear height. The yield-lodging relationship was affected by genotype and location. We concluded that maize yield could be enhanced by optimizing plant density, applying DHEAP and cultivar selection, but climatic and environmental differences of locations should be considered.  相似文献   

Molecular mapping of soybean seed tocopherols in the cross ‘OAC Bayfield’ × ‘OAC Shire’          下载免费PDF全文

Eric J. Shaw  Istvan Rajcan 《Plant Breeding》2017,136(1):83-93

Soybean (Glycine max [L.] Merr.) is cultivated primarily for its protein and oil in the seed. In addition, soybean seeds contain nutraceutical compounds such as tocopherols (vitamin E), which are powerful antioxidants with health benefits. The objective of this study was to identify molecular markers linked to quantitative trait loci (QTL) that affect accumulation of soybean seed tocopherols. A recombinant inbred line (RIL) population derived from the cross ‘OAC Bayfield’ × ‘OAC Shire’ was grown in three locations over 2 years. A total of 151 SSR markers were polymorphic of which a one‐way analysis of variance identified 42 markers whereas composite interval mapping identified 26 markers linked to tocopherol QTL across 17 chromosomes. Individual QTL explained from 7% to 42% of the total phenotypic variation. Significant two‐locus epistatic interactions were identified for a total of 122 combinations in 2009 and 152 in 2010. The multiple‐locus models explained 18.4–72.2% of the total phenotypic variation. The reported QTL may be used in marker‐assisted selection (MAS) to develop high tocopherol soybean cultivars.  相似文献   

Effect of eleutheroside B/E on proliferation and expression of PPARγ and TGF-β1 in HBZY-1 cells cultured with high glucose     

WU Jian-hua  DU Yin-xiang  HUANG Qiang 《园艺学报》2015,31(3):511-517

AIM: To explore the effects and mechanism of eleutheroside (ETS) B or E on the proliferation of HBZY-1 cells treated with high glucose. METHODS: The HBZY-1 cells were cultured under high glucose condition. The 4th generation of HBZY-1 cells was used for determining the optimal cell density, which was consistent with the growth regulation curve of the cells. The cells were divided into 6 groups: low glucose (LG) group, high glucose (HG) group, high glucose plus ETS-B/E (low dose, medium dose and high dose) groups, and high glucose plus losartan (LTG) group. After all cells were treated with the corresponding drugs at 24 h, 48 h and 72 h, the inhibitory rate of the proliferation was measured, and the expression of TGF-β1 and PPARγ was detected by immunocytochemistry and Western blotting. RESULTS: The best cell density was 2 000 cells/well, which was complied with the basic rules of the cell growth, and high glucose significantly promoted the HBZY-1 cell proliferation. At each time point, the inhibitory effects of ETS-B/E were significantly different between HG group and LTG group on the proliferation of the HBZY-1 cells (P<0.05). The expression of TGF-β1 was significantly inhibited, and the expression of PPARγ was significantly promoted by ETS-B/E (P<0.05). ETS-E showed stronger effect than ETS-B (P<0.05) in a concentration- and time-dependent manner. CONCLUSION: ETS-B/E significantly inhibits the proliferation of HBZY-1 cells under high glucose condition by decreasing TGF-β1 expression and promoting PPARγ expression.  相似文献