Expression,Purification and Activity Evaluation of Mb1230 Gene of Mycobacterium bovis     

LI Ming  JIA Hong  XIN Ting  GUO Xiao-yu  YUAN Wei-feng  HOU Shao-hua  HOU Qiang  GAO Xin-tao  WU Jing  DONG Rui-kai  YANG Hong-jun  LIU Lai-xing  ZHU Hong-fei 《中国畜牧兽医》2015,42(10):2544-2550

To explore the value of Mb1230 protein of Mycobacterium bovis in the diagnosis of bovine tuberculosis,we obtained the Mb1230 gene by PCR and constructed recombinant plasmid pET-22b-Mb1230.Recombinant Mb1230 protein was obtained by IPTG induction and purified by affinity chromatography.The activity of the recombinant protein was evaluated by TST test,IGRA test and indirect ELISA.The size of the recombinant protein matched with the theoretical value proved by SDS-PAGE;Western blotting result showed that the recombinant protein could react with mouse anti-His antibody,and had specific band;The results of TST test,IGRA test and indirect ELISA test also showed the recombinant protein had antigenic activity.The results indicated the recombinant protein Mb1230 had good B cell and T cell activity,so,it had the potential application in the diagnosis of bovine tuberculosis.  相似文献   

激素对奶牛乳腺上皮细胞乳脂肪合成的调控作用     

曹越  李大彪 《中国畜牧兽医》2021,48(8):2778-2786

乳脂肪是高质量的天然脂肪，其可为人类提供营养和能量，在各种膳食脂肪和油类中，是最容易被消化吸收的。乳脂肪是在乳腺中由从头合成或外源摄取的脂肪酸与甘油酯化形成的一种脂类物质，其含量的高低关系着牛奶品质的优劣和乳制品的加工特性。在奶牛的泌乳周期中，乳腺泌乳功能受多种因素影响，其中内分泌腺分泌的多种激素对奶牛乳腺上皮细胞（BMECs）乳脂的合成具有积极的调控作用。综上所述，作者介绍了氢化可的松、催乳素、胰岛素和生长激素4种泌乳相关激素对BMECs乳脂肪合成的调控机理，即从乳脂合成适宜的激素添加量、激素对乳脂球形态的影响方面初步阐释其调控作用，并从乳脂合成的关键酶及转录因子、激素对乳脂合成相关基因表达量方面深入阐释其作用机理，旨在为研究泌乳相关激素对奶牛乳腺内乳脂肪合成的调控机理提供参考。  相似文献   

Effect of Psammaplin A on in vitro Development of Bovine Aging Oocytes     

WANG Huaqing  GAO Qingshan  JIANG Wenjie  ZHAO Yuhan  JIN Qingguo  LI Yinghua  XU Yongnan 《中国畜牧兽医》2019,46(2):497-503

To investigate the effect of histone deacetylation inhibitor Psammaplin A (PsA) on the development of bovine aging oocytes in vitro,oocytes were randomly divided into control group,aging group and 50 mmol/L PsA treated aging group (PsA group).Immunofluorescence staining and JC-1 were used to detect the blastocyst rate of bovine oocytes after parthenogenetic activation,the number of cells in blastocysts,apoptosis,reactive oxygen species (ROS),glutathione (GSH) and mitochondrial membrane potential intensity of embryos.The results showed that the blastocyst rate of the aging group was significantly lower than that of PsA and control groups (P<0.05).The blastocyst rate of PsA group was not significantly different from that of control group (P>0.05).The number of cells in the blastocysts of control group and PsA group were significantly higher than that of aging group (P<0.05).The number of cells in the blastocysts of PsA group was not significantly different from that of control group (P>0.05).The apoptosis rate in aging group was significantly higher than that of control and PsA groups (P<0.05),the apoptosis rate of PsA group was significantly higher than that of control group (P<0.05).The GSH level of MⅡ oocytes in aging group was significantly lower than that of control and PsA groups (P<0.05).There was no significant difference in GSH level between control and PsA groups (P>0.05).The ROS level of the embryos in aging group was significantly higher than that of control and PsA groups (P<0.05).The ROS level in PsA group was significantly higher than that of control group (P<0.05).The mitochondrial membrane potential of early embryos of aging group 4-8 cells was significantly lower than that of control and PsA groups (P<0.05).The mitochondrial membrane potential intensity of control group was significantly higher than that of PsA group (P<0.05).In summary,PsA could effectively delay the aging of bovine oocytes and improve the quality of oocytes.  相似文献   

Future Directions for Research in Infectious Bovine Keratoconjunctivitis     

《Veterinary Clinics of North America: Food Animal Practice》2021,37(2):371-379

  相似文献   

Sequence Analysis and Prokaryotic Expression of NSP1 Gene of Bovine Rotavirus UK Strain     

RAN Xu-hua  CAO Si  ZHANG Yao  WEI Xiao-man  WEN Xiao-bo  NI Hong-bo  ZHU Zhan-bo 《中国畜牧兽医》2015,42(5):1104-1109

Rotavirus is a major cause of acute diarrhea in both many kinds of young animals and children under 5 years old.Rotavirus NSP1, a 55 ku RNA binding protein, is the product of gene 5, which can subvert innate immune responses and be one of virulent determinant factors.According to the sequence in GenBank, specific primers targeting to NSP1 gene were designed and the gene was amplified by RT-PCR, following by being cloned into the pET-28a(+) vector.It showed that the full length of NSP1 gene was 1 473 bp, encoding 491 amino acids.The NSP1 shared the highest identity with WC3 strain.The recombinant protein was induced in E.coli Rosetta(DE3) by IPTG and was analyzed by SDS-PAGE and Western blotting.The results revealed that NSP1 recombinant protein existed in the form of inclusion body with the molecular weight of 55 ku.The purified recombinant protein could be recognized by His-tag antibody.This study laid the foundation for further research on the relationship between the intracytoplasmic location of NSP1 protein and its activity.  相似文献   

苦参碱对奶牛乳腺上皮细胞增殖、凋亡及抗氧化能力的影响     

卢金霞  何芳  冯峰 《西北农林科技大学学报(自然科学版)》2018,46(7):1-6

【目的】探究苦参碱对体外培养的奶牛乳腺上皮细胞(BMECs)增殖、凋亡及抗氧化能力的影响。【方法】利用含0(A组),25(B组),50(C组),75(D组)和100μg/mL(E组)苦参碱的培养基培养奶牛乳腺上皮细胞。通过四甲基偶氮唑盐(MTT)法检测BMECs活性,采用流式细胞仪(AnnexinV/PI双染法)检测苦参碱对BMECs凋亡的影响,并检测苦参碱对BMECs抗氧化酶活性及丙二醛(MDA)含量的影响,采用real-time PCR对BMECs中Caspase-3、p53、STAT1和SOCS3基因的相对表达量进行检测。【结果】用药5d时,低质量浓度(25和50μg/mL)苦参碱对BMECs增殖具有促进作用,高质量浓度(75和100μg/mL)苦参碱对细胞增殖具有抑制作用;B~E组BMECs的凋亡率均极显著高于A组(P0.01);B~E组BMECs培养上清液中NO和乳酸脱氢酶(LDH)水平明显高于A组。B~E组BMECs的过氧化氢酶(CAT)活性均比A组高,其中C组极显著高于A组(P0.01);B~E组的谷胱甘肽过氧化物酶(GSH-Px)活性均极显著高于A组(P0.01),E组的超氧化物歧化酶(SOD)水平极显著高于A组(P0.01),各组MDA含量无显著性差异。与A组相比,苦参碱上调了B~E组BMECs中Caspase-3、p53、STAT1和SOCS3基因的相对表达量。【结论】低质量浓度苦参碱能够促进BMECs增殖,高质量浓度苦参碱则会抑制BMECs增殖;不同质量浓度苦参碱均可提高BMECs的抗氧化能力,其中50μg/mL苦参碱提高BMECs抗氧化能力的效果最明显。  相似文献   

Whole-genome sequence of a novel Chinese cyprinid herpesvirus 3 isolate reveals the existence of a distinct European genotype in East Asia     

《Veterinary microbiology》2015,175(2-4):185-194

Cyprinid herpesvirus 3 (CyHV3), also known as koi herpesvirus (KHV), can be subdivided primarily into European and Asian genotypes, which are represented by CyHV3-U or CyHV3-I and CyHV3-J, respectively. In this study, the whole genome sequence of a novel Chinese CyHV3 isolate (GZ11) was determined and annotated. CyHV3-GZ11 genome was found to contain 295,119 nucleotides with 52.9% G/C content, which is highly similar to those of published CyHV3-U, CyHV3-I, and CyHV3-J strains. With reference to CyHV3-U, CyHV3-I, and CyHV3-J, CyHV3-GZ11 was also classified into 164 open reading frames (ORF), which include eight repeated ORFs. On the basis of the 12 alloherpeviruses core genes, results from phylogenetic analysis showed that CyHV3-GZ11 had closer evolutionary relationships with CyHV3-U and CyHV3-I than with CyHV3/KHV-J, which were also supported by genome wide-based single nucleotide substitution analysis and the use of a series of developed molecular markers. This study was the first to reveal the presence of a distinct European CyHV3 genotype in East and Southeast Asia at a whole genome level, which will evoke new insights on exploring the origin, evolution, and epidemiology of the virus.  相似文献   

口蹄疫疫苗对牛的应用效果调查     

余金霞 《畜牧兽医科学(电子版)》2021,(4):17-18

牛口蹄疫是传染性疾病,为研究青海省峰堆乡口蹄疫疫苗的免疫效果,于2018—2019年对该乡的7个村开展牛口蹄疫疫苗免疫效果调查,共调查牛634头,7个村总体的口蹄疫合格率为100.00%和97.56%,最低合格率为81.63%和88.63%。为更好地控制该类疾病发生流行,提升免疫效果,该文探讨口蹄疫疫苗免疫失败原因,并制定了相应的预防措施。  相似文献   

白藜芦醇通过激活去乙酰化酶1/腺苷一磷酸激活的蛋白激酶信号通路影响牛脂肪细胞凋亡     

成海建  游伟  靳青  刘倚帆  万发春  宋恩亮  刘桂芬  谭秀文  张相伦  刘晓牧 《动物营养学报》2017,29(12):4398-4407

本试验旨在研究植物提取物白藜芦醇(RES)对牛皮下脂肪细胞凋亡率以及去乙酰化酶1(SIRT1)/腺苷一磷酸激活的蛋白激酶(AMPK)信号通路关键因子的mRNA和蛋白质表达量的影响。选取18月龄鲁西黄牛的皮下前体脂肪细胞,在细胞分化的第0天,更换为RES浓度分别为0(对照)、100、200和400μmol/L的培养液处理48 h,每组设3个重复。应用Hoechst33342染色检测细胞凋亡的形态学变化,流式细胞分析仪检测细胞凋亡率,荧光定量PCR(q PCR)和免疫印迹试验(Western-blot)分别检测SIRT1/AMPK信号通路上的关键因子SIRT1、A MPKα、叉头转录因子1(Fox O1)、B细胞淋巴瘤/白血病-2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、促凋亡蛋白Bcl-2相关X蛋白(Bax)的mRNA和蛋白质表达量,并利用油红O染色鉴定脂肪细胞。结果表明,与对照组相比:不同浓度RES处理后的牛皮下脂肪细胞凋亡率均极显著增高(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3、Bax的mRNA表达量均显著或极显著提高(P0.05或P0.01),Bcl-2的mRNA表达量极显著降低(P0.01);不同浓度RES处理后的SIRT1、AMPKα、caspase-3的蛋白质表达量均显著或极显著提高(P0.05或P0.01);200和400μmol/L RES处理后,Fox O1的mRNA和蛋白质表达量显著或极显著提高(P0.05或P0.01),Bax的蛋白质表达量极显著提高(P0.01),Bcl-2的蛋白质表达量极显著降低(P0.01)。100μmol/L RES处理后时,Fox O1的mRNA和蛋白质表达量以及Bcl-2、Bax的蛋白质表达量均与对照组差异不显著(P0.05)。由此可见,RES通过激活SIRT1/AM PK信号通路,同时激活通路下游的Fox O1,促进了牛皮下脂肪细胞的凋亡,为通过营养调控技术降低牛皮下脂肪沉积提供了一定的理论基础。  相似文献   

Detection of Ureaplasma diversum in the upper airways of Australian feedlot cattle     

RJ Barnewall  IB Marsh  PMV Cusack  F Galea  N Sales  JC Quinn 《Australian veterinary journal》2023,101(6):254-257

Bovine respiratory disease (BRD) exerts a major impact on the beef cattle industry nationally and worldwide, with a range of aetiological factors impacting its pathogenesis. Previous research has focussed on an increasing number of bacteria and viruses that have been shown to play a role in eliciting disease. Recently, additional agents have been emerging as potential contributors to BRD, including the opportunistic pathogen Ureaplasma diversum. To determine if U. diversum was present in Australian feedlot cattle and if that presence was linked to BRD, nasal swabs were collected from a cohort of 34 hospital pen animals and compared to 216 apparently healthy animals sampled contemporaneously at feedlot induction and again after 14 days on feed at an Australian feedlot. All samples were subjected to a de novo polymerase chain reaction (PCR) assay targeting U. diversum in combination with other BRD agents. U. diversum was detected at a low prevalence in cattle at induction (Day 0: 6.9%, Day 14: 9.7%), but in a significantly greater proportion of cattle sampled from the hospital pen (58.8%). When considering the presence of other BRD-associated agents, co-detection of U. diversum and Mycoplasma bovis was most common in hospital pen animals receiving treatment for BRD. These findings suggest that U. diversum may be an opportunistic pathogen involved in the aetiology of BRD in Australian feedlot cattle, in combination with other agents, with further studies are warranted to identify if a causal relationship exists.  相似文献