苏云金芽孢杆菌cry1I基因沉默的研究

分析了苏云金芽孢杆菌(Bt)cry1和cry1I基因的保守区,在cry1类基因的下游和cry1I基因的上游设计了一对通用引物,用于检测cry1I与cry1类基因的连锁现象.从鉴定的142株Bt分离株中,PCR扩增发现71株出现约1.4kb的阳性带,说明这一现象广泛存在于Bt菌株中.在此基础上用pGEM-T Easy载体克隆了菌株Bt07和J8中的连锁序列,序列分析表明,Bt07中是cry1I基因与cry1Db基因连锁,间隔区为504bp;J8中是cry1Ia基因与cry1Ac基因连锁,间隔区为506bp.进一步比较间隔区序列,同源性达93.7%,同时含有多个原核生物终止序列,且未发现启动子序列,揭示了这两个菌株中cry1I基因沉默的成因.     

二价铜离子对牛正常朊蛋白二级结构的影响

研究了二价铜离子对牛朊蛋白的二级结构的影响.牛朊蛋白(BoPrPc)的小分子(BoPrPcS)和大分子(BoPrPcL)经初步纯化后,在含Cu2+的碱性溶液中氧化,用反相柱进行氧化态和还原态的BoPrPcL和BoPrPcS的分离.质谱分析证明氧化态的Bo-prPcL和BoPrPcS分子质量分别为24.064,15.819ku.用远紫外圆二色谱(Far-UVCD)分析显示,BoPrPcS在208和222m处出现两个负峰,为典型的α-螺旋型结构;而Bo-PrPcL在222nm处负峰减小.与未经氧化的BoPrPcL的质谱和CD结果比较后发现,氧化后的BoPrPcL在八肽重复区结合了6个Cu2+,其β-折叠含量明显增加.而Cu2+对BoPrPcS二级结构基本无影响.BoPrPcL结合Cu2+的特性及其二级结构的相应变化可能与细胞内抗Cu2+毒性与氧化压力有关.     

X13—1菌体蛋白对小鼠免疫功能的影响

研究高蛋白含量的Fusarium菌株X13－1的最佳生长条件,并从菌丝体中获得了高达47％的菌体蛋白,再用热休克方法使其中RNA的含量降至2％以下,使之符合WHO标准,通过动物免疫实验证明,这种部分纯化的菌体蛋白可以增强小鼠的免疫功能。     

GFP标记的GDNF重组腺病毒载体的构建和体内外表达

通过构建绿色荧光蛋白(GFP)和胶质细胞源性神经营养因子(GDNF)共表达的腺病毒载体,讨论其在治疗脊髓和脑有关神经系统疾病的潜在应用价值,了解腺病毒载体在大鼠体内的分布及在体外分泌外源基因产物能力,并对其生物安全性作了初步考察,为GDNF重组腺病毒载体在临床和基础研究中的应用作了一定准备．     

人类基因组计划和我们的生活

对于基因的科学研究,是从发现基因的传递规律开始的,1965年,一位奥地利神父--孟德尔--发现并发表了有关基因传递的规律.20世纪中叶,科学家发现了基因--遗传物质的基本结构,并提出了模型,即我们在中学教科书里读过的"DNA双螺旋模型".20世纪70年代,人类破天荒第一次真正能够拿出基因来做试验,即建立了可以称之为"遗传工程"或"分子克隆"的技术,并第一次用拼凑基因的方法建立了人为的生物,尽管还不完全,还很简单,科学家却用一些基因生产出了"遗传工程"的产品--基因产品.     

Study of apolipoprotein E genetic polymorphism in patients with atherosclerotic cerebral infarction

Objective: To explore the frequency and significance of ApoE gene polymorphisms in Chinese patients with atherosclerotic cerebral infarction (ACI). Methods: Polymerase chain reaction and gene sequencing, single nucleotide polymorphisms of ApoE gene were used to analyze 33 cases of patients with ACI and 35 controls. Results: The frequencies of ApoE gene single nucleotide polymorphisms 465C/G, 462C/G and 451delC in the ACI group were significantly higher than those in the control group (P<0.05). The prevalence of polymorphism 486G/T in the control group was significantly higher than that in the ACI group (P=0.011). Conclusions: 465C/G,462C/G and 451delC polymorphisms might be associated with ACI.486GT allele might have protective effect on the pathogenesis of ACI.     

Construction of ice nucleation active <Emphasis Type="Italic">Enterobacter cloacae</Emphasis> for control of insect pests

Ice nucleation active(INA)bacteria are the most potent heterogeneous ice nuclei in nature,which have become an important biological resource for diverse applications.Many researches have proved that INA bacteria can raise the supercooling points(SCPs)of insect pests,then reduce their cold hardiness.However,INA bacteria‘s inefficient colonization on the surface or in the guts of insects,and the high incidence of frost injury induced by their release hampered the application of INA bacteria in controlling insect pests in agricultural fields.In this study,we constructed a recombinant plasmid mob-Tn5-iceA with the ability of broad-host-range conjugal mobilization and integration of the ina gene of icaA into chromosomal DNA of many gram-negative bacteria by Tn5 transposition.In addition,Ent.cloacae strains stably carrying iceA and expressing high ice nucleation activity(INA),even in the absence of antibiotic pressure,were constructed through conjugal mobilization and Tn5 transposition.Ent,cloacae strains have benn reported to be able to efficently colonize in the guts of insects,but have weak plant epiphytic ability.Therefore,these transgenic Ent.Cloacae may be promising candidates for control of insect pests in agricultural fields.     

中华绒螯蟹卵巢新基因EJO6的全长cDNA克隆和序列分析

利用RACE技术从中华绒螫蟹卵巢获得了新基因EJO6(Eriocheir japonica ovary gene 6,EJO6)的全长cDNA序列(GenBarnk 检索号：AYl85922)。该cDNA序列长度为1250bp,开放阅读框为690bp,编码229个氨基酸。根据氨基酸序列计算的相对分子质量和等电点分别为24270和11．9。同时用生物信息学的方法对该基因的结构和功能进行了初步分析。     

Recombinant PsbF from Synechococcus sp. PCC 7002 forms β:β homodimeric cytochrome b559

All organisms with oxygenic photosynthesis contain two photosystems:photosystem Ⅰ(PSⅠ)and photo-systemⅡ(PSⅡ),The minimal photosystem Ⅱ particles which are photochemically active contain three subunits:D1,D2 and cytochrome b559 (Cyt b559),The function of Cyt b559 remains unclear,We have successfully overxpressed the psbF gene,encoding the β subunit of Cyt b559,from a marine cyanobacterium Synechoccous sp.PCC 7002 as a fusion gene and obtained a redox-active from of Cyt b559,When the N-terminal GST protein of the fusion gene product was removed with thrombin ,the PsbF protein was still redox-active,suggesting that the recombinant PsbF can form dimer in Escherichia coli.The absorption spectra of either the oxidized from or the reduced form of both GST fusion protein and the purified PsbF dimer and the difference Spectra between the two forms are the same as that of the Cyt b559 isolated from the higher plants .Redox titration analysis of recombinant PsbF showed that the mid-point redox potential of the recombinant Cyt b559 was approximately 50 mV, which is close to the low potential of Cyt b559 ,The results are helpful to the understanding of locatlization and function of Cyt b559 on thylakoid membranes.Ⅰ