籼粳质核互作雄性不育恢复基因等位性研究

籼粳质核互作雄性不育恢复基因等位性研究严菊强,沈圣泉,薛庆中（浙江农业大学农学系,杭州３１００２９）ＴＧ７是由ＣＰＳＬＯＩ７／明恢６３经花药培养产生的广谱型广亲和恢复系,对珍汕９７Ａ等籼型和７６２７Ａ等粳型质核互作雄性不育系均有恢复能力,并带有与ＣＰ...     

环状芽孢杆菌C—2几丁酶基因在大肠杆菌中的表达

对已克隆的环状芽孢杆菌Ｃ－２的几丁酶基因片段所作的亚克隆分析表明,该几丁酶基因位于１．７ｋｂ Ｐｓｔ Ｉ－Ｓｔｙ Ｉ片段 。ＣｈｉＩ基因在大肠杆菌ＪＭ１０７,ＤＨ５α,ＸＬ１－ｂｌｕｅ,ＴＧ－１等菌株中均能表达,但表达水平不同,其中ＪＭ１０７的表达活性最高,其胞外几丁酶活性与供体菌Ｃ－２菌株的胞外酶活性几乎相当。     

GL—7—ACA酰化酶在大肠杆菌中的分泌表达

利用来自假单胞菌的ＧＬ－７－ＡＣＡ酰化酶的信号肽和表达元件基因片段构建了ＧＬ－０７－ＡＣＡ酰化酶的分泌型高表达质粒ｐＴｒｃＣＡ１Ｓ和ｐＫＫＣＡ１Ｓ,其中ｐＴｒｃＣＡ１Ｓ为ＩＰＴＧ诱导型质粒,ｐＫＫＣＡ１Ｓ为组成型质粒。ｐＴｒｃＣＡ１Ｓ和ｐＫＫＣＡ１Ｓ转入受体菌ＴＧ１中都可高表达ＧＬ－７－ＡＣＡ酰化酶基因并将表达产物转运到周质空间,完整细胞酰楷酶比活力分别为２３．９单位每克菌体和１８．３单位每克菌体     

人I型胶原基因第一内含子调节转录的研究

人Ｉ型胶原α１（Ｉ）链（ＣＯＬＩＡ１）基因内含子序列在不同细胞内有不同的转录调节活性,报道了含人ＣＯＬＩＡ１基因内含子Ｉ不同区段（＋５４４～＋８５５和＋８２０～＋１０９３）的重组质粒ｐＳＣＥＰ－ＣＡＴ和ｐＳＣＩＰ－ＣＡＴ的构建并转染人胚肌腱成纤维细胞和Ｔｃａ８１１３舌癌细胞,地高辛标记抗ＣＡＴ－ＥＬＩＳＡ检测结果显示：ｐＳＣＥＰ－ＣＡＴ在两种细胞均获表达;ｐＳＣＩＰ－ＣＡＴ在人成纤维细胞未表达,但     

STUDIES ON THE PATTERN OF MEGASPOROGENESIS AND MICROTUBULAR CYTOSKELETON CHANGES IN CYMBIDIUM SINENSE

ＳＴＵＤＩＥＳＯＮＴＨＥＰＡＴＴＥＲＮＯＦＭＥＧＡＳＰＯＲＯＧＥＮＥＳＩＳＡＮＤＭＩＣＲＯＴＵＢＵＬＡＲＣＹＴＯＳＫＥＬＥＴＯＮＣＨＡＮＧＥＳＩＮＣＹＭＢＩＤＩＵＭＳＩＮＥＮＳＥ￥Ｓ．Ｙ．ＺｅｅＸ．Ｌ．Ｙｅ（１ＢｏｔａｎｙＤｅｐａｒｔｍｅｎｔ,Ｕｎｉ...     

人组织型纤溶酶原激活剂（t—PA）组合突变体FrGGl在CHO细胞中的…

为了得到ｔ－ＰＡ组合突体ＦｒＧＧｌ在ＣＨＯ细胞中的高效表达,将表达质粒筛选基因启动子上游的增强子（ｅｎｈａｎｃｅｒ）去除,构建了ＦｒＧＧｌ真核表达质粒ｐＺＬＦｒＧＧＩ。酶切线化后,采用大剂量ＤＮＡ电击介导法,转染ｄｈｆｒ基因缺陷型中仓鼠卵巢细胞系（ＣＨＯ－ｄｈｆｒ）。氨甲喋呤（ＭＴＸ）筛选转染细胞,混合加压,挑选克隆,在１×１０＾－７ｍｏｌ／ＬＭＴＸ压力下,获得表达水平达１５００～２５００ＩＵ／１     

大鼠卵巢绒毛膜促性腺激素受体在CHO中的表达和扩增

本文报道了利用二氢叶酸还原酶放大系统将大鼠ＬＨ／ｈＣＧ受体（记为Ｆ－ｈＣＧＲ）及其胞外肽段（记为Ｔ－ｈＣＧＲ）在中国苍鼠卵巢细胞（ＣＨＯ）中的表达。ＳＤＳ－ＰＡＧＥ分析表明,Ｆ－ｈＣＧＲ为一条蛋白质带,其表观分子量为９２ｋｄ,而Ｔ－ｈＣＧＲ为３５ｋｄ和３７ｋｄ两条带。表达受体对其配基ｈＣＧ表现出高的亲合力,Ｆ－ｈＣＧＲ的解离常数为７×１０－９ｍｏｌ／Ｌ,Ｔ－ｈＣＧＲ为６．４×１０－９ｍｏｌ／Ｌ。表达Ｆ－ｈＣＧＲ的转染ＣＨＯ细胞可结合１２５Ｉ－ｈＣＧ,而表达Ｔ－ｈＣＧＢ者不结合１２５Ｉ－ｈＣＧ。这提示Ｆ－ｈＣＧＲ主要存在于细胞质膜表面上。表达Ｆ－ｈＣＧＲ的转染ＣＨＯ细胞能刺激。ｃＡＭＰ的形成,而表达Ｔ－ｈＣＧＲ者不能刺激ｃＡＭＰ形成。免疫荧光定位结果表明,Ｔ－ｈＣＧＲ主要分布于质膜的细胞质侧以及胞内其他一些细胞器膜上。用免疫亲和层析可以得到纯化的Ｔ－ｈＣＧＲ。     

c-fos基因在内皮素-1促肺泡Ⅱ型细胞表面活性物质合成中的作用

应用反义技术探讨ｃ－ｆｏｓ基因ＥＴ－１调控肺泡Ⅱ型细胞（ＡＴⅡ）表面活性物质（ＰＳ）合成的胞内信号转导中的作用,结果显示：（１）内皮素－１（ＥＴ－１）可提高ＡＴⅡ细胞的＾３Ｈ－胆碱掺入。（２）蛋白激酶Ｃ（ＰＫＣ）激活剂ＰＭＡ可使ＡＴⅡ细胞的＾３Ｈ－胆碱掺入量增加,ＰＫＣ抑制剂Ｈ７可抑制ＥＴ－１的促ＰＳ合成效应。（３）ＥＴ－１和ＰＭＡ可显著提高Ｆｏｓ蛋白表达量,Ｈ７和ｃ－ｆｏｓ反义寡核苷酸（ＯＤＮ）     

家蝇体内卵对CAT的摄入和传代的观察

家蝇体内卵对ＣＡＴ的摄入和传代的观察ＩＮＴＡＫＩＮＧＡＮＤＴＲＡＮＳＭＩＳＳＩＯＮＯＦＩＮＪＥＣＴＥＤＣＡＴＧＥＮＥＢＹＨＯＵＳＥＦＬＹＥＧＧＳ关键词ＣＡＴ基因,家蝇,腹腔注射,基因转移ＫｅｙｗｏｒｄｓＣＡＴ－ｇｅｎｅ,Ｈｏｕｓｅｆｌｙ,Ａｂｄｏｍｉ...     

利用脊髓灰质炎病毒5‘NCR和RNA聚合酶基因可提高外源基因表达

将含脊髓灰质炎病毒（ＰＶ）ＲＮＡ聚合酶的不同长度基因片段克隆到载体ｐＳＧ５质粒上,分别构建了４个表达ＲＮＡ聚合酶的质粒。体外转录实验证明,ｐＳＧ５－ＰＯＬ１．９９和ｐＳＧ５－ＰＯＬ２．０３质粒转染细胞的提取物促进了特异的ＲＮＡ转录,表明两质闰可表达ＲＮＡ聚合酶。将ＰＶ的５’ＮＣＲ序列插在载体ｐＧＲＥＥＮ ＬＡＮＴＥＲＮ－１的ＣＭＶ启动子下游,构建了ｐＧＲＥＥＮ ＬＡＮＴＥＲＮ－１－５’ＮＣＲ质粒;     

高等植物成花基因的研究

高等植物的成花可分为两个阶段：由茎顶端分生组织转变成花分生组织和花器官的形成。前者主要受成花计时基因的控制,而后一阶段主要由植物的同源（异型）框基因调控。本文综述了近年来对植物成花调控基因的研究,并着重对第一阶段成花基因的功能、它们间的相互作用和特点进行了总结。 Abstract:Plant flower can be divided into two phases——from stem apex meristem tissue into flower meristem tissue and floral apparatus.The flower time genes control the flower development and the homologous genes control flower apparatus identify.This paper summarizes recent studies on plant flower and emphasizes on the first phase flower control genes,theirs interaction and function,characteristic of the homologous genes.     

Putative novel Bradyrhizobium and Phyllobacterium species isolated from root nodules of Chamaecytisus ruthenicus

In this study, the diversity and the phylogenetic relationships of bacteria isolated from root nodules of Chamaecytisus ruthenicus growing in Poland were investigated using ERIC-PCR fingerprinting and by multilocus sequence analysis (MLSA). Two major clusters comprising 13 and 3 isolates were detected which 16S rRNA gene sequencing identified as Bradyrhizobium and Phyllobacterium. The results of phylogenetic analysis of individual and concatenated atpD, gyrB and recA gene sequences showed that the studied strains may represent novel species in the genera Bradyrhizobium and Phyllobacterium. In the phylogenetic tree based on the atpD-gyrB-recA concatemers, Bradyrhizobium isolates were split into two groups closely related to Bradyrhizobium algeriense STM89^T and Bradyrhizobium valentinum LmjM3^T. The genus Phyllobacterium isolates formed a separate cluster close to Phyllobacterium ifriqiyense LMG27887^T in the atpD-gyrB-recA phylogram. Analysis of symbiotic gene sequences (nodC, nodZ, nifD, and nifH) showed that the Bradyrhizobium isolates were most closely related to Bradyrhizobium algeriense STM89^T, Bradyrhizobium valentinum LmjM3^T and Bradyrhizobium retamae Ro19^T belonging to symbiovar retamae. This is the first report on the occurrence of members of symbiovar retamae from outside the Mediterranean region. No symbiosis related genes were amplified from Phyllobacterium strains, which were also unable to induce nodules on C. ruthenicus roots. Based on these findings Phyllobacterium isolates can be regarded as endophytic bacteria inhabitating root nodules of C. ruthenicus.     

A Dual Role for Microbial Pathogen-Derived Effector Proteins in Plant Disease and Resistance

Many proteins from plant pathogens affecting the interaction with the host plant have dual functions: they promote virulence on the host species and they function as avirulence determinants by eliciting defense reactions in host cultivars expressing the appropriate resistance genes. In viruses all proteins encoded by the small genomes can be expected to be essential for viral development in the host. However, in different plants surveillance systems have evolved that are able to recognize most of these proteins. Bacteria and fungi have specialized pathogenicity and virulence genes. Many of the latter were originally identified through the resistance gene-dependent elicitor activity of their products. Their role in virulence only became apparent when they were inactivated or transferred to different microbes or after their ectopic expression in host plants. Many microbes appear to maintain these genes despite their disadvantageous effect, introducing only few mutations to abolish the interaction of their products with the plant recognition system. This has been interpreted as been indicative of a virulence function of the gene products that is not impaired by the mutations. Alternatively, in particular in bacteria there is now evidence that pathogenicity was acquired through horizontal gene transfer. Genes supporting virulence in the donor organism's original host appear to have traveled along. Being gratuitous in the new situation, they may have been inactivated without loss of any beneficial function for the pathogen.     

Phylogenetically diverse groups of Bradyrhizobium isolated from nodules of tree and annual legume species growing in Ethiopia

Bacteria belonging to the genus Bradyrhizobium nodulate various leguminous woody plants and herbs, including economically important crops such as soybean, peanut and cowpea. Here we analysed 39 Bradyrhizobium strains originating from root nodules of the leguminous trees and crops Acacia saligna, Faidherbia albida, Erythrina brucei, Albizia gummifera, Millettia ferruginea, Cajanus cajan, Vigna unguiculata and Phaseolus vulgaris, growing in southern Ethiopia. Multilocus sequence analyses (MLSA) of the 16S rRNA, glnII, recA, gyrB and dnaK genes and the ITS region grouped the test strains into seven well-supported genospecies (I–VII), six of which occupied distinct positions excluding all hitherto defined Bradyrhizobium species. Analyses of the nodA, nodC and nifH genes suggested different evolutionary history of the chromosomal and symbiosis-related genes. Our study corroborates earlier findings that Ethiopia is a hotspot for rhizobial biodiversity, justifying further search for novel strains from this region and calling for intensified research on the ecology and biochemistry of these organisms.     

利用相似致病菌中保守的致病菌特有基因预测新的毒力相关基因

目的:在致病机制相似的致病菌中寻找保守的致病菌特有基因,预测新的毒力相关基因。方法:首先选取致病机制相似的致病菌EHEC与EPEC,利用本实验室构建的包含115 152条致病菌特有基因片段的数据库进行本地Blast,得到致病菌特有基因,对致病菌特有基因在相似致病菌中的保守性进行分析,得到新的可能的毒力相关基因。结果:在6株EHEC菌中找到95条保守的致病菌特有基因,其中大部分为已知的毒力相关基因,还有许多可能的毒力相关基因;在9株相似致病菌(EHEC、EPEC)中找到10条保守的致病菌特有的蛋白基因,其中9条为已知的致病相关基因,1条为可能的致病相关基因。结论:应用本方法可以发现新的毒力基因,为后续对致病菌致病机制的实验研究奠定了基础。     

Conservation of the RD-BF-C2 organization in the pig MHC class-III region: mapping and cloning of the pig RD gene

The RD gene, named after the arginine (R) and aspartic acid (D) repeat in the central part of its protein, was initially mapped in the mouse H-2S subregion between C4 and BF. It was later mapped in the same position in the human MHC and here we show it is also conserved in the pig MHC class III region, close to the complement BF gene. A pig RD genomic clone was isolated from a γ-phage library. Hybridizations on genomic DNA separated with pulsed field gel electrophoresis identified common 220kb Nrul, 130 kb EagI and 200 kb Mlul bands for RD, BF and C2. The RD gene has also a 17 kb Kpnl and 11 kb Sad fragment in common with BFbut not with C2. The close linkage of the RD and BF genes was further established by hybridization of BF to a genomic γ-phage clone also containing the RD gene. This genomic RD clone overlaps with a γ -phage clone previously isolated and containing the complete BF gene and the 3' part of C2. The distance between RD and BF is about 6 kb. The junction between the two complement genes BF and C2 was sequenced and the BF 5' promoter region, overlapping the 3' noncoding region of C2, was compared with that of the human BF promoter. The overall homology was about 80% and all but one identified promoter elements were found in the same position in both genes. The results obtained demonstrate the RD-BF-C2 organization is strongly conserved between human, mouse and pig. No polymorphisms were detected in either the RD gene or in the BF promoter region using polymerase chain reaction and restriction fragment polymorphism analysis.     

杜氏盐藻分子生物学最新进展及展望

杜氏盐藻是一种无细胞壁的单细胞双鞭毛真核藻类,是一种十分重要的藻类资源。过去对杜氏盐藻的研究多集中在形态学、耐盐机理及β-胡萝卜素等方面,近年来,随着藻类基因工程的快速发展,本研究课题组及国内外在杜藻盐藻分子生物学方面做了大量工作,现就杜氏盐藻在这一领域的研究进展进行综述,主要是重要功能基因的克隆与分析、杜氏盐藻调控序列的研究以及杜氏盐藻作为宿主表达外源基因等。     

HCMV增殖机制研究进展

人类巨细胞病毒(human cytomegalovirus,HCMV)感染在人群中广泛存在,病毒在感染细胞内增殖会对宿主细胞进行多水平调节和干扰,从而导致各种宏观疾病。本文对HCMV复制周期的基因表达时序、HCMV-DNA复制相关蛋白质及其编码基因、HCMV-DNA复制特点和HCMV必需基因、非必需基因及抑制基因等进行综述。