Detection,cloning and expression of bone morphogenetic protein-1 from human osteosarcoma cell lines     

CHEN Dong-ying  ZHU Quan-sheng  LIU Chao  QIU Ju-shi 《园艺学报》2003,19(6):751-756

AIM:To manufacture recombinant protein of the highly conserved domain in human bone morphogenetic protein-1(BMP-1) using gene engineering methods as antigen for making wide spectrum antibody to BMP-1.METHODS:We analyzed the gene sequences and protein structures of BMP-1 and its related proteins, and chose a highly conserved fragment as target gene. Total RNA was prepared from human osteosarcoma cell line Saos-2, then the target gene was amplified with RT-PCR. The PCR product was cloned into prokaryotic expression vector pMAL c2 to get recombinant vector BMP-1(322-588aa)-pMAL c2. After transforming the recombinant plasmid into DH5-alpha and screening, several prositive clones were got for sequencing. Finally the transformed cells was induced with IPTG to get fusion protein.RESULTS:The BMP-1 gene fragment was successfully cloned into vector pMAL c2, and was able to express efficiently with IPTG inducement. The amount of expressed fusion protein is about 66%-72% in total volume of bacterial proteins.CONCLUSIONS:The recombinant protein contains several key domains(2 CUB domains and 1 EGF domain), which are shared by BMP-1 and its related proteins. Specific wide spectrum antibody to human BMP-1 and its related proteins may be generated with this recombinant protein antigen.  相似文献   

Effect of heat shock precondition on reperfusion arrhythmia in rats     

XIE Zi-an  HUANG Yuan-wei  HU Shen-jiang  XIA Qiang  SHEN Yue-liang 《园艺学报》2002,18(5):449-453

AIM:To investigate the effect of the heat shock response on the reperfusion arrhythmias(RAs) and the possible mechanism involved. METHODS:Fifty-five Sprague Dawley rats were randomly divided into 2 groups: the heat shock group (group H,n=29) and the control group (group C,n=26). The rats in group H were preconditioned with heat shock 24 hours before, and that in group C were not. The hearts of 16 rats in group H and 16 in group C were exercised and mounted on a non-circulating Langendorff perfusion apparatus and perfused retrogradely with modified K-H buffer and mimic ischemia/reperfusion was applied. Additionally, conventional intracellular microelectrode techniques were used for recording such electrophysiological parameters as resting potential(RP), action potential amplitude(APA), over shot(OS), maximum depolarization velocity(Vmax) of the hearts of other 13 rats in group H and 10 in group C. RESULTS:①Prior heat stress significantly decreased reperfusion arrhythmia. ②The amount of CK release in the effluent in group H was much less than that in group C. ③Myocardial HSP70 content was elevated significantly in group H. ④Heat stress significantly increased myocardial anti-oxydases activity and decreased lipid peroxydative products. Additionally, heat stress significantly reduced the Vmax of action potential. It indicated that rapid Na⁺ channel of papillary muscles may be inhibited by heat shock. The degree of change of Vmax after ischemia in H group was significantly less than that in group C. And the time of reperfusoin with Tyrode's solution till the action potential appeared as large as that before perfusion with mimic ischemic solution is shorter in group H than in group C. CONCLUSION:Heat shock pretreatment markedly reduces ischemia/reperfusion-induced injury of heart and ventricular arrhythmias in rats and this effect may be associated with the inhibition of rapid Na⁺ channel of papillary muscles by heat shock and the increase in myocardial HSP70 and anti-oxydase activity.  相似文献   

Effect of protein kinase C on the expression of myocardial HSP70 mRNA during myocardic ischemic preconditioning in rabbits     

LU Yan-zhen  WANG Yong-jin  WANG Xiao-li  WANG Li-min 《园艺学报》2002,18(4):406-409

AIM:To study the relationship between myocardial HSP70 and PKC during myocardial ischemic preconditioning(IPC). METHODS: PKC inhibitor, polymyxin B(PMB) and PKC activator, phorbol 12-myristate 13-acetate(PMA) were applied to the models of myocardial ischemia/reperfusion in vivo and in vitro in rabbits, respectively, and the ventricular functions, PLA₂ in the serum, and the expression of mycardial HSP70 mRNA were examined. RESULTS:IPC decreased PLA₂ activity, improved the left ventricular function and increased the expression of myocardial HSP70 mRNA. Howerer, all these effects of IPC could be blocked by PMB. Interestingly, PMA mimicked IPC with attenuating the injuries of cardial myocytes and increasing myocardial HSP70 mRNA expression. CONCLUSION:PKC is involved in the myocardial HSP70 expression in case of ischemic preconditioning.  相似文献   

Screening for Differential Gene Expression in Atropa belladonna Leafy Gall Induced Following Rhodococcus fascians Infection     

El Hassan Nouar  Danny Vereecke  Koen Goethals  Mondher Jaziri  Marie Baucher 《European journal of plant pathology / European Foundation for Plant Pathology》2003,109(4):327-330

To better characterise at the molecular level the nature of plant responses to infection by Rhodococcus fascians PCR-based differential display patterns of Atropa belladonna leafy gall (LG) and non-infected plant tissues were compared. Six differentially expressed genes were identified and their altered expression was confirmed by RT-PCR. Three of them corresponded to up-regulated genes which encode proteins involved in plant defence. The three remaining cDNA fragments which correspond to down-regulated genes in LG, encoded proteins with similarity to a multicystatin, a miraculin and a methallothionein-like protein, respectively. Upon elimination of the bacteria from infected plant tissue, the expression of up-regulated genes was maintained, whereas expression of down-regulated genes resumed suggesting a potential role of these up-regulated genes in plant growth and development.  相似文献   

Effect of Epstein-Barr virus latent membrane protein 1 on expression of P53 protein in nasopharyngeal carcinoma cells     

ZHANG Su-zhen  HUANG Pei-chun  XU Yong  CHEN Jin  CAI Kang-rong  HUANG He 《园艺学报》2004,20(9):1646-1649

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Protective effect of HSP₇₀ on injuried myocardial cells induced by H₂O₂     

ZHANG Wei-hua  WANG Li-na  LI Quan-feng  XU Chang-qing 《园艺学报》2004,20(3):402-406

AIM: To investigate the role of heat-shock protein 70(HSP₇₀) in the protection of myocardial cells against ischemic injury.METHODS: Myocardial cells were cultured in vitro. HSP₇₀ was induced by hyperthermia. H₂O₂-injured myocardial cells were divided into different groups. Flow cytometry, DNA Ladder and biochemistry methods were employed to observe the myocardial cells of different groups.RESULTS: Immunohistochemistry showed hyperthermia induced the up-regulation of HSP₇₀ in myocardial cells. Apoptotic rate, activity analysis of cytochrome C and succinic dehydrogenase in H₂O₂-injuried and HSP₇₀-protected groups were obviously different. Electron micrograph shomed hyperthermia alliviated myocardial cell injury induced by H₂O₂. CONCLUSION: HSP₇₀ delays apoptosis and protects against H₂O₂-induced myocardial cell injury.  相似文献   

中国苜蓿育成品种遗传多样性及亲缘关系研究   总被引：1，自引：0，他引：1  

郭江波  赵来喜 《中国草地学报》2004,26(1):9-13

以苜蓿的种子贮藏蛋白的多态性为研究指标,对中国14个审定登记的苜蓿育成品种的遗传多样性及亲缘关系进行了研究。研究结果表明:蛋白单体的分子量介于21.2～85.1kD之间,蛋白单体电泳图谱多态性品种间无显著的特异性;在参试的14个育成品种中,品种内基因多样性平均值(HS)为0.3136,总基因多样度(HT)为0.3741,基因分化系数(GST)为0.162;以λ=3.700为标准划分,14个育成品种划分为六个组群,各组群划分结果主要与参试品种的亲本来源有关,而与其种植地、育种地关系不大。  相似文献   

大豆和百脉根古老原核基因的全基因组鉴定与比较分析     

韦兴燚  李昱  刘文献  金小煜  闵学阳  张正社  NdayambazaBoniface  王彦荣 《草业学报》2018,27(11):49-57

在漫长的进化过程中,高等真核生物从原核生物中获取了大量基因,对这些基因进行鉴定分析,可为研究高等植物系统演化及基因组水平比较分析提供理论依据。为了进一步解析大豆和百脉根古老原核基因在其基因组中所起的作用及进化关系,本研究利用生物信息学技术对大豆和百脉根基因组古老原核基因进行全基因组鉴定,并对其特征及功能进行了比较分析。研究结果表明,大豆(40.6%)古老原核基因的占比高于百脉根(33.9%),且多分布于内共生细胞器线粒体、叶绿体中。此外,古老原核蛋白质的结构域在大豆和百脉根基因组中的分布相似,表明其在大豆和百脉根进化过程中具有较高的共线性和保守性。通过GO注释发现大豆与百脉根中古老原核蛋白主要分布在膜系统、细胞、细胞组分中。在分子功能上,大豆古老原核蛋白更多地参与代谢和发育过程,这可能与其接受了更多的人为选育有关。在生物过程中,古老原核蛋白更多参与催化反应和结合反应,主要以酶的形式在发挥作用。通过比较分析,本研究揭示了古老原核蛋白在大豆和百脉根中进化的独特模式,可为其他豆科植物基因组分析提供一定理论基础。  相似文献   

Effects of continuous low dose infusion of lipopolysaccharide on inflammatory responses,milk production and milk quality in dairy cows          下载免费PDF全文

L. T. Ning  G. Z. Dong  C. Ao  D. G. Zhang  K. Erdene  F. Q. Zhang  J. Wen  T. L. Zhang 《Journal of animal physiology and animal nutrition》2018,102(1):e262-e269

The objective of this study was to evaluate the effects of continuous low dose infusion of lipopolysaccharide (LPS) on inflammatory responses and milk production and quality in lactating dairy cows. Eight Holstein cows were assigned to two treatments in a cross‐over experimental design. Cows were infused intravenously either with saline solution or with saline solution containing LPS from Escherichia coli O111:B4 at a dose of 0.01 μg LPS/kg body weight for approximately 6 hr each day during a seven‐day trial. The clinical symptoms and milk production performance were observed. Milk samples were analysed for conventional components, fatty acids and amino acids. And jugular vein and mammary vein plasma samples were analysed for concentrations of cytokines and acute phase proteins. LPS infusion decreased feed intake and milk yield. An increase in body temperature was observed after LPS infusion. LPS infusion also increased plasma concentrations of interleukin‐1β, serum amyloid A, LPS‐binding protein, C‐reactive protein and haptoglobin. LPS infusion decreased the contents of some fatty acids, such as C17:1, C18:0, C18:1n9 (trans) and C18:2n6 (trans), and most amino acids except for methionine, threonine, histidine, cysteine, tyrosine and proline in the milk. The results indicated that a continued low dose infusion of LPS can induce an inflammatory response, decrease milk production and reduce milk quality.  相似文献   

Identification of a class of CFEM proteins containing a new conserved motif in Fusarium oxysporum     

《Physiological and Molecular Plant Pathology》2015

Fusarium oxysporum (Fo) is an important soil-borne fungus that cause huge economic losses worldwide. Fungal-specific Common in Fungal Extracellular Membrane (CFEM) proteins are known to be involved in some important physiological processes associated with pathogenicity. To date, few Fo CFEM proteins have been characterized. The recent publication of several genomes of Fo has allowed us to conduct a genome-wide comparison analysis of CFEM proteins in Fo. In this study, we identified CFEM proteins for 12 different Fo formae speciales(f. sp) and obtained an average of 16 CFEM proteins for each Fo. The Fo CFEM proteins were classified into three groups (groups 1–3) according to structural features. Importantly, we identified a new conserved motif containing about 50 amino acids (DR motif) in group 1 members. CFEM proteins containing DR motif (CFEM_DR proteins) are remarkably conserved among Fo, and their number is greater in Fo compared with other fungi. We found the expansion of CFEM_DR proteins in Fo can be attributed to the segmental duplications in the genomes. Expression analysis revealed that CFEM_DR genes had a higher expression level in mycelium than conidia, and their expressions increased dramatically in the host roots at 3 days post inoculation, indicating that Fo CFEM_DR genes have roles while colonizing and infecting their hosts.  相似文献