用体外定向进化的方法对EPSPS合酶基因草苷膦抗性机制的研究

为了寻找EPSP合酶中某些与草苷膦抗性相关的位点,利用体外定向进化技术,对可变盐单胞菌HT7的EPSP合酶基因,荧光假单胞菌G2的EPSP合酶基因以及按植物偏爱密码子合成G2的EPSP合酶基因,进行DNA shuffling,通过EPSP合酶缺陷型菌株E.coil ER2799的功能互补筛选,获得了7个草苷膦抗性有较大变化的EPSP合酶突变体.     

含内蛋白子融合表达载体的构建与人神经营养因子-3的初步表达

为了构建一个含蛋白质剪接元件的表达载体 ,将 3.38kb的 p Ex Sec 的多克隆位点 (Eco R /Bam H )处插入一个含多个酶切位点的 49bp寡聚核苷酸。然后将 p MYB12 9中 intein- CBD基因片段移插入上述经改造的p Ex Sec I中 ,构建成含内蛋白子的表达载体 p Ex IC。根据人神经营养因子 - 3(h NT- 3)的已知 DNA序列 ,设计含 NdeI/Xho I酶切位点的引物 ,用 PCR法从人全血总 DNA中扩增 h NT- 3,再插入 p Ex IC载体中 ,构建成 p Ex IC- h NT- 3重组质粒。经转化后 ,工程菌 E.coli BL 2 1(DE3) /p Ex IC- h NT- 3在 L B培养基中获得融合表达 ,根据 intein的自剪切原理 ,在还原剂 DTT作用下 ,初步获得了约 14k D的 h NT- 3目的蛋白。     

有限群的可补子群的相关性质

考虑有限群的极小子群和Sylow子群的可补性质对群结构的影响.设F是包含全体有限超可解群的群系,G是有限群,M>1是G的正规子群,且G/M∈F,证明:如果对M的任一极小子群H,H∩F*(GF)均在G中可补,则G∈F.     

HDTV视频同步与消隐时序控制信号的CPLD设计

ATSC HDTV视频输出采用了SMPTE的相应标准．将像素计数与行计数相结合，采用统计行分类编码的算法对SMPTE274M、SMPTE296M、SMPTE293M和SMPTE295M标准的同步与消隐时序控制信号进行了多平台的仿真分析，并在ALTERA公司的EPM7128上最终实现了本设计．     

A hypersensitive site in hsp70 chromatin requires adjacent not internal DNA sequence

Nuclease-hypersensitive sites in chromatin exist at the 5' side of many eukaryotic genes. To gain some understanding of the molecular basis of these hypersensitive sites, we have now examined the pair of sites upstream of the Drosophila hsp70 gene in a series of plasmids that contain deletions in the hypersensitive region and have been transformed into yeast cells. Hypersensitive sites 5' to a Drosophila hsp70 gene are preserved when this gene is introduced into yeast by transformation. We find that a yeast strain containing a plasmid in which the deletion extends through the first hypersensitive site still displays the normal pair of hypersensitive sites, so DNA sequences over which the first hypersensitive site is centred are not required for hypersensitivity at this position and the site can form over a foreign DNA sequence juxtaposed against this deletion end point. Deletions progressing further into the region bracketed by the pair of 5' hypersensitive sites eliminate the first hypersensitive site and alter the downstream site. We propose that the hypersensitive sites are generated through the binding of a protein that renders flanking sequences more accessible to nucleases, perhaps by preventing normal chromatin packaging.     

关于半正规的几类群

利用子群的半正规性讨论了几类有限群的结构,得到如下主要结果:(l)极大子群超可解的有限群当其极大子群的极小子群半正规时,它不是超可解群就是如下三种群之一:(I)p~αq~β阶内-Abel群,p(?)q-1;(Ⅱ)p~(α+β)r(?)阶群,α≥2,β≥0,p~β=│φ(G)│,p~(α-1)||r—1,α~((?)~α+β)=c_1~(?)=c_2~(?)=…=c_(?)~(?)=1,c_ic_j=c_jc_i,i,j=1,2,…,p,c_(?)~(?)=c_(i+1),i=1,2,…,p-1,c_(?)~(?)=c_1~(?),t(mod r)指数p~(α-1);(Ⅲ)D_(2_q)型群;(2)极大子群可解的非Abel有限单群当其二次极大子群的极小子群半正规时,G恰为A_5.     

Crystal structure of opsin in its G-protein-interacting conformation

Opsin, the ligand-free form of the G-protein-coupled receptor rhodopsin, at low pH adopts a conformationally distinct, active G-protein-binding state known as Ops*. A synthetic peptide derived from the main binding site of the heterotrimeric G protein-the carboxy terminus of the alpha-subunit (GalphaCT)-stabilizes Ops*. Here we present the 3.2 A crystal structure of the bovine Ops*-GalphaCT peptide complex. GalphaCT binds to a site in opsin that is opened by an outward tilt of transmembrane helix (TM) 6, a pairing of TM5 and TM6, and a restructured TM7-helix 8 kink. Contacts along the inner surface of TM5 and TM6 induce an alpha-helical conformation in GalphaCT with a C-terminal reverse turn. Main-chain carbonyl groups in the reverse turn constitute the centre of a hydrogen-bonded network, which links the two receptor regions containing the conserved E(D)RY and NPxxY(x)(5,6)F motifs. On the basis of the Ops*-GalphaCT structure and known conformational changes in Galpha, we discuss signal transfer from the receptor to the G protein nucleotide-binding site.     

人巨细胞病毒临床低传代病毒株UL97 基因的克隆与分析

分离人巨细胞病毒（HCMV）临床低传代GZ02病毒株,并根据GenBank提供的实验室标准病毒株HCMV AD169磷酸转移酶基因UL97 DNA序列及有关文献设计引物,从HCMV GZ02病毒株基因组DNA中通过PCR扩增UL97基因,并克隆至pGEM3Z质粒载体．重组质粒经测序鉴定,发现HCM VGZ02病毒株UL97基因序列保守区域与HCMV AD169 UL97长度完全一致,但发生G205A、G761A、T823C、T1173C、T1282C、T1334C、T2106C等7个位点的碱基突变,涉及到编码氨基酸E69K,S275P,C428R和F445S位点发生改变。     

新型冠状病毒膜蛋白的生物信息学分析

为了研究新型冠状病毒膜蛋白(SARS-CoV-2 M蛋白)结构及性质.基于生物信息学分析M蛋白质基因结构、二级结构和三级结构、翻译后的修饰和进化历程.结果表明,M蛋白为疏水性蛋白,其基因编码区长度为669bp,编码222个氨基酸;M蛋白启动子区内不存在甲基化位点,存在17潜在的转录因子结合位点;其二级结构以无规则卷曲和...     

南方鲇卵子发生的超微结构研究

1996年 4月至 1998年 4月 ,以嘉陵江北碚段收集和人工繁殖饲养的处于不同发育阶段的南方鲇雌鱼性腺为材料 ,利用透射电镜研究了其卵子的发生过程 ,共分为卵原细胞期、卵黄发生前期、卵黄发生期、卵黄发生后期和成熟卵子 5个阶段 .卵原细胞的核质比例大 ,有中央大核仁 ,胞质中细胞器少 .处于卵黄发生前期过程中的卵母细胞 ,核膜波曲 ,核仁物质外排明显 ,有线粒体云的存在 .线粒体云主要由线粒体组成 ,同时还有内质网、高尔基体及泡状结构等组分 .卵黄发生期的主要标志是皮层泡的出现 ,皮层泡由高尔基体产生 ,到卵黄发生旺盛阶段 ,核仁物质大量外排 ,排出的核仁物质在胞器、膜性小泡或直接在胞质中沉积形成卵黄小板 .线粒体、内质网、高尔基体、核糖体等胞器丰富、功能活跃 ,其中 ,线粒体尤为突出 .卵母细胞的胞饮作用也特别明显 .卵黄发生后期 ,卵黄物质占据卵母细胞的大部分 ,各种细胞器减少 ,功能减退 ,卵黄合成能力大大减弱 .到成熟卵子 ,卵黄物质几乎充满整个细胞 ,除少部分细胞器维持卵子的功能外 ,其余的均沉积卵黄物质形成卵黄小板     

一种自剪切内含肽重组酿酒酵母表达优化研究

自剪切内含肽纯化系统在大肠杆菌中已经得到很好的应用。为了在真核生物中应用该系统,以酿酒酵母为宿主,p HR质粒为表达载体,构建微型内含肽ΔI-CM intein酿酒酵母表达系统。以猪免疫球蛋白Ig G的Fc domain为亲和标签,绿色荧光蛋白gfp为报告蛋白,在酿酒酵母GPD强启动子作用下,表达出融合的Fc-intein-gfp蛋白。检测发现融合序列在起始密码子ATG前加入一段Kozak序列有利于gfp表达。在此基础上,将微型内含肽ΔI-CM intein序列替换成密码子优化的ΔI-CMintein-O序列,显著促进gfp蛋白的表达量,初步实现了ΔI-CM intein融合蛋白的高效表达。为新型自剪切内含肽酿酒酵母表达系统的建立和重组蛋白高效纯化的实现奠定了基础。     

Detection of genetically modified ingredients in seed samples from Heilongjiang soybean field

A total of 210 soybean samples collected from different areas of Heilongjiang Province were analyzed by PCR to detect the presence of RR (Roundup Ready) soybean ingredient. Results showed that CaMV35S promoter was not detected in all samples. CP4-EPSPS gene was amplified in 13 samples, but all of them were proved to be false positives in restriction endonuclease digestion analysis of PCR products. Further analysis by nested PCR indicated that there were no RR soybean ingredients in the samples. Besides, the amplified fragments of 0xl by CP4-EPSPS primers were sequenced, and the results confirmed again the fact that this fragment was not from CP4-EPSPS gene.     

具有给定分支数森林的最小能量图

将图G的能量E(G)定义为图G的特征多项式所有特征根的绝对值之和.F_n,q是顶点个数为n,分支个数为q的森林的集合. 对于给定的n和q,给出F_n,q中具有最小能量的图.     

The lambda phage att site: functional limits and interaction with Int protein

Site specific integrative recombination of bacteriophage lambda involves unequal partners. The minimal phage att site is composed of approximately 240-base pairs and four distinct binding sites for Int protein, at least three of which are crucial for function. This 'donor site' recombines efficiently with a smaller 'recipient site' that lacks the extensive interactions with Int protein.     

Construction of a site-specific, deletion-frameshift mutation in an essential gene of bacteriophage phiX174.

By in vitro methods, we have deleted 80 nucleotides from a preselected site in the coding sequence of gene G of bacteriophage phiX174. This mutant (Gdelta/FS ZH1) also carries a--2 frameshift. The mutation is lethal, but mutant virus can be stably maintained on host strains bearing plasmids carrying phiX gene G.     

极小子群完全条件可换的有限群

设F是一个子群闭的局部群系，具有下列性质：极小非F-群可解，且它的F-上根是一个Sylow子群。如果群G的任意4阶循环子群在G中完全条件可换，且G的任意极小子群包含于G的F-超中心内，那么G是一个F-群。     

Molecular mechanism of the inhibitory effect of methylation outside the recognition sequence of restriction endonuclease Pvu II on its cleavage activity

In 1991, we found that methylation outside the Pvu II recognition sequence could partially inhibit its cleavage activity. To clarify the molecular mechanism, three plasmids with different methylation states were constructed. Then, together with the original one, four plasmids were digested with different amounts of Pvu II. Results show that methylation on both sites results in 90 % inhibition; moving the methylated site one base further away decreases the inhibitory effect to about 30 %; with the adjacent dam methylation site eliminated, the inhibitory effect disappears. The data suggest that the inhibition of cleavage activity caused by outside methylation is not “all or none”, and the degree of inhibition is dependent on the position and the number of methylated bases.     

拟阵Q_6与W~4可F-线性表示的构造

对拟阵 Q6与W4可F-线性表示的构造进行了研究.用E(G)在R上的链群F0(G,R)表示G的圈拟阵M(G);用松弛拟阵M的极小圈超平面X的方法得到拟阵M′.得到主要结果为:(1)用链群表示了M(K4),M(W4);(2)用松弛极小圈超平面的方法从M(K4)构造了Q6,从M(W4)构造了W4,找出了W4可线性表示的所有域F.     

有限群的正规p-补的一些结论

有限群的极小子群在群论研究中有很重要的地位。文章探讨极小子群对有限群的p-幂零性,并得到:设P是群G的Sylowp-子群,满足Ω1(P∩F(G))≤Z∞(G),如果NG(Z(P))有一个正规p-补,那么G有一个正规p-补;若G还没有与A4同构的主因子,则G有一个正规p-补。