High efficient expression of human endostatin in E.coli and its antiangiogensis activity

AIM: To examine the expression of human endostatin in E.coli, produce its fusion protein antibody and observe its biological activity. METHODS: Endostatin gene was amplified by polymerase chain reaction,recombined with plasmid vector pGEX-2T and induced expression with IPTG.The protein activity was tested by endothelial cell proliferation inhibitory assay.Inclusion body crudely purified was used to generate polyclonal antibody to detect its expression at mouse's liver and kidney etc. RESULTS: The protein expressed was 20kD after digestion by thrombin,it appeared the anti-angiogenesis activity and Western blotting indicated the expression of endostatin in liver and kidney of mouse. CONCLUSION: The successful expression of human endostatin and the preparation of polycolonal antibody indicated its potential application in anti-angiogenesis therapy and diagnosis tumors.     

两系杂交粳稻产量性状优势形成分析

采用程氏指数对两系杂交组合亲本及其杂种F1进行分类,对杂种F1产量性状与亲本及杂种F1程氏指数,杂种F1产量性状与父本产量性状进行相关分析。结果表明,偏粳×粳、偏粳×偏粳的组合杂种F1为偏粳类型。杂种F1每穗总粒数表现与双亲程氏指数差呈显著正相关,同时也受到父本性状值的影响。     

黄土高原东北缘退耕坡地土壤养分和容重空间变异特征研究

以山西广灵退耕6年的典型坡地为代表,在实地调查和取样分析的基础上,使用经典统计学和地统计学方法研究了黄土高原东北缘退耕荒草地中土壤养分与容重的空间变异特征.结果表明:研究区土壤容重较高、养分含量较低,土壤较为贫瘠,且土壤容重具有弱变异而土壤养分具有中等程度变异;同时,所研究土壤性质的合理采样数从多到少依次为全磷>有机质>速效磷>碱解氮>速效钾>全氮>容重;另外,该区土壤养分和容重的空间分布可用球状模型进行较好的拟合,而且其空间变化主要是人为等随机性因素作用的结果.     

以分子标记辅助选择育成抗条纹叶枯病水稻新品种“武陵粳1号”

条纹叶枯病是21世纪以来江苏省最重要的水稻病害之一。食味品质较优的主栽品种武育粳3号因高度感染该病而种植面积锐减。本研究以镇稻88为抗条纹叶枯病毒病基因Stvb-i的供体亲本,采用回交育种策略,改良武育粳3号的抗条纹叶枯病性能。在连续回交和自交过程中,以紧密连锁的双侧分子标记对Stvb-i进行“前景”选择,同时对后代与轮回亲本遗传背景的相似程度进行分子标记辅助“背景”选择,在短期内育成抗条纹叶枯病的“武育粳3号”,命名为“武陵粳1号”。新品种保持了原品种的基本农艺性状、丰产性、稳产性和优异的食味品质,并大幅度提高了其条纹叶枯病抗性水平,在江苏省多点抗性鉴定试验中的平均病株率仅为4.4%,极显著低于原品种(53.2%)。     

Effect of exogenous hydrogen sulfide on lipophagy in mouse primary hepatocytes

AIM: To evaluate the effect of exogenous hydrogen sulfide (H₂S) from GYY4137 on lipophagy in mouse primary hepatocytes. METHODS: The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion method were divided into 4 groups: the cells in control group were incubated with normal medium; the cells in model group were incubated with 1.2 mmol/L oleic acid (OA) for 48 h; the cells in H₂S group or propargylglycine (PAG) group were incubated with 1.2 mmol/L OA for 48 h followed by serum-free phenol red-free RPMI-1640 medium which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h. The cells were collected to conduct immunofluorescence staining of LC3 and photography under fluorescence microscope, phase-contrast microscope or transmission electron microscope. The protein expression of LC3-Ⅰ/Ⅱ in the hepatocytes was determined by Western blot. RESULTS: In contrast with the model group, the fluorescent particles of LC3, the protein expression of LC3, the number of autophagic lysosome and vacuoles in hepatocytes in H₂S group increased. CONCLUSION: In steatosis hepatocytes, exogenous H₂S promotes the lipophagy.     

高产优质水稻新品种9香优007的选育

9香优007系铜仁鑫天地农业发展有限公司利用自育恢复系铜恢007与自育不育系9香A杂交选育的新品种.主要特点是:米质优、抗性强、丰产稳产性好.具有较好的推广前景.     

菜田土壤养分空间变异特征

采用传统统计和地统计相结合的方法,对55 hm2菜田耕层土壤养分空间变异特征进行了研究。结果表明,土壤养分主要限制因子是N、P、K和Zn,不同土壤养分的变异情况各不相同。土壤养分速效含量一般呈斑块状分布;土壤NO3--N、P、K和Zn含量在空间分布上具有明显的空间相似性。土壤NO3--N、P、K含量分别与种菜历史及肥料用量(N、P2O5和K2O)之间均呈极显著的正相关。     

红椿SRAP反应体系优化及引物筛选

[目的]优化相关序列扩增多态性(SRAP)体系内的不同组分,建立适用于红椿SRAP分子标记的反应体系,并进一步从SRAP引物组合中筛选出稳定、多态性好的引物组合,为红椿遗传多样性研究奠定试验基础。[方法]针对SRAP-PCR反应体系中5个因素各设置8个水平,先利用单因素试验确定浓度梯度,后在确定的梯度范围内选定4个水平,按照正交试验L16(45)进行优化,结合正交直观分析法和新复极差法对各因素进行优化筛选。[结果]确定最优体系为总体系25μL,模板DNA 25 ng,上下游引物各0.3μmol·L~(-1),Taq DNA聚合酶1U,Mg2+2.5 mmol·L~(-1),d NTP 0.3 mmol·L~(-1)。利用稳定的SRAP-PCR体系,从1 505对SRAP引物组合中筛选出30对优质引物组合。[结论]通过不同种源红椿基因组DNA的重复验证,获得了稳定清晰、多态性较强的扩增条带,表明所确定的最优体系稳定可靠,适用性较强,可用于不同种源红椿遗传多样性研究的后续实验。     

副猪嗜血杆菌PCR检测方法的建立与初步应用

针对副猪嗜血杆菌(Haemophilus parasuis,HPS)16 S rRNA序列设计1对特异性引物,能扩增出821 bp的特异性DNA片段,并据此建立了快速准确鉴定副猪嗜血杆菌PCR方法,临床试验证明该方法具有很好的特异性和敏感性.13份疑似病料检测结果表明,PCR检测结果与传统生化鉴定结果符合率为100％.结果表明已成功建立了副猪嗜血杆菌PCR检测方法,并可应用于临床副猪嗜血杆菌的检测.