动物的的变态

变态是动物学中一个较重要的专用名词,有关内容在中学课本也多处涉及到。现择要介绍一点动物变态的知识,供动物学教学参考。何谓动物的变态动物由于外在和内在的原因,个体形态发生变化,这叫变态。但动物学所讲的变态,是狭义地从发生学角度理解,即胚胎不直接转变为成体,而是在后期发育过程中,先形成形态、生理、生态方面特殊的幼体,行独立生活和生长,以后在某阶段发生急剧变化,转变为成体。青     

活的不可培养的细菌的研究进展

活的不可培养微生物(VBNC)即一些微生物明显地丧失了可培养的特性,但是保留了自身原有的代谢活力,并且在一定条件下,又可以回复到可培养的状态。从VBNC细菌的诱导条件、生物学特性和检测方法3个方面对VBNC细菌研究进展做一综述。     

真核细胞的基因的组织

一、真核细胞基因的基本结构 1.转录单位: 从已知的数十种基因的顺序,可得出一个具有功能的基因的共同规律,在基因5’端-25至-75区,有CCAAT和TATAAA区(后者又称ATA box或Hogness box),相当于促进子区(Promotor),为体外转录所必需。     

高频率的波动对于种子的萌发和植物的发育的影响

本文主要是以理论和试验来说明音波对植物的生长发育和种子萌发所起的影响。在农业实践上音波所起的作用,据现在所知:有缩短植物成熟期,加速萌芽和增强植物的生长发育等。这一些非但具有理论和实践上的意义,同时在今後把物理科学应用到农业科学中开辟了极广阔的前程。     

由吸附的缩氨酸诱导的细胞膜的形变

研究了由一系列相互平行的吸附在细胞膜上的缩氨酸引起的膜的弹性形变,以及膜对缩氨酸的包裹行为,得到膜的平衡方程,用它可以来处理大尺度的形变,弯曲能量、吸附能量和弹性形变的相互竞争导致膜对缩氨酸发生从不吸附到部分吸附乃至完全包裹的结构转变．在膜的形变很小的时候,可以得到系统能量的解析解。     

远古的人们是怎样认识自己的起源的

人是从那里来的? 回答这个问题,你也许会说这有什么困难——人是从古猿变来的;甚至你还会进一步说,在这个从猿到人的转变过程中,劳动起着决定性的作用。然而这个现在看来比较明了的道理,恰是经历了多么漫长的认识过程才达到的呵!现在让我们首先来谈谈,远古的人们是怎样认识自己的起源的。最初的原始人可能还想不到自己的起源在人类诞生的最早时期,“最初的、从动物界分离出来的人,在一切本质方面是和动物本身一样不自由的”(恩格斯:《反杜林论》),这些最初的原始人为艰苦     

敲除pckA基因的结核杆菌引起的免疫反应的研究

研究结核杆菌pckA基因编码的磷酸烯醇型丙酮酸羧激酶(PEPCK)诱导机体产生的保护性免疫反应。用敲除pckA基因的牛结核杆菌BCG和野生型BCG分别感染小鼠,取肝、肺、脾进行病理分析,并进行脾细胞培养,检测CD4 、CD4 /CD8 、细胞因子IFNI-γI、L-12和TNF等。用敲除pckA基因的BCG感染的小鼠比野生型BCG感染的小鼠体内产生的结核结节少且不典型,炎性程度低。野生型BCG感染的小鼠脾脏内的CD4 T细胞和CD4 /CD8 、细胞因子IFN-γ、IL-12、TNF均明显高于敲除pckA基因BCG感染的小鼠。pckA基因为结核杆菌生长所必需,其编码产物PEPCK能够刺激机体产生免疫反应,是一种很好的疫苗候选分子。     

分离的蚕豆细胞核的RNA聚合酶活力的研究

利用Triton X-100对叶绿体膜的作用,可快速地从蚕豆幼叶制备较纯净的细胞核,它具有较高的RNA聚合酶活力。比较了两种分离核的方法,证明利用匀浆法制备的核具有较高的活力。核活力与发育时期有关系,茎端和第1对幼叶的核活力显著高于第2和第3对叶片的核活力。此外,核活力明显地受反应液内锰离子的抑制。     

霸王的原生质体培养的研究

目的：为利用原生质体融合技术转移霸王抗旱基因。方法：采用酶解法分离霸王原生质体,比较了霸王子叶和愈伤组织游离原生质体的产量和活力,不同渗透压和起始密度对原生质体分裂频率的影响。结果：愈伤组织游离的原生质体产量和活力均高于子叶,原生质体产率可达2.4×106个/g.FW,活力达89%。采用液体浅层培养,在附加2,4-D（2mg/L）、6-BA（1.0mg/L）、2%蔗糖和甘露醇（0.4mol/L）的DPD培养基中,原生质体分裂频率最高,达68.6%。转移到附加2-iP（3mg/L）、KT（1.0mg/L）、6-BA（1.0mg/L）的分化培养基上,获得2个再生苗。结论：采用酶解法游离霸王愈伤组织,可获得高活力和高分裂频率的霸王原生质体。     

Identification of vaccinia promoters by heterologous expression of hepatitis B surface antigen in mouse cells infected by recombinant vaccinia viruses

DNA fragments preceding open reading frames in a conserved segment of the vaccinia virus genome (Plucienniczak A., et al. (1985) Nucleic Acids Res. 13, 985-998) were cloned into plasmids upstream of the S gene of the hepatitis B virus encoding the surface antigen (HBsAg). Recombinant vaccinia virus obtained after insertion of these constructs into the thymidine kinase gene were used to infect mouse 1D cells. HBsAg was assayed in cellular supernatants. A strong promoter was thus identified in a 295 bp fragment preceding the coding region of the 147 kDa subunit of the vaccinia RNA polymerase.     

Identification of vaccinia promoters by heterologous expression of hepatitis B surface antigen in mouse cells infected by recombinant vaccinia viruses

DNA fragments preceding open reading frames in a conserved segment of the vaccinia virus genome (Plucienniczak A., et al. (1985) Nucleic Acids Res. 13, 985–998) were cloned into plasmids upstream of the S gene of the hepatitis B virus encoding the surface antigen (HBsAg). Recombinant vaccinia virus obtained after insertion of these constructs into the thymidine kinase gene were used to infect mouse 1D cells. HBsAg was assayed in cellular supernatants. A strong promoter was thus identified in a 295 bp fragment preceding the coding region of the 147 kDa subunit of the vaccinia RNA polymerase.     

Attempted therapeutic immunization in a chimpanzee chronic HBV carrier with a high viral load

BACKGROUND: We previously reported successful therapeutic immunization in a chimpanzee having a relatively low viral load, which was immunized with recombinant plasmid hepatitis B surface antigen (HBsAg) DNA and boosted with recombinant HBsAg encoding canarypox virus. In the present study, we attempted to confirm these findings in an animal with a high virus load. METHODS AND RESULTS: We tested three immunization strategies successively over a 3-year period. In the first of these, we administered four monthly injections of DNA encoding HBsAg + PreS2 + hepatitis B core antigen (HBcAg) + DNA encoding interleukin (IL)-12, (given 3 days later), and boosted with canarypox expressing all of the above HBV genes 6 months after initial immunization. No reduction in viral load was observed. In the second trial, we administered lamivudine for 8 weeks, and then began monthly DNA-based immunization with plasmids expressing the above viral genes; however, viral loads rebounded 1 week after termination of lamivudine therapy. In a third trial, we continued lamivudine therapy for 30 weeks and immunized with vaccinia virus expressing the above viral genes 18 and 23 weeks after the start of lamivudine therapy. Again viral loads rebounded shortly after cessation of lamivudine treatment. Analysis of cell-mediated immune responses, and their avidity, revealed that DNA-based immunization produced the strongest enhancement of high avidity T-cell responses, while recombinant vaccinia immunization during lamivudine therapy enhanced low avidity responses only. The strongest low and high avidity responses were directed to the middle surface antigen. CONCLUSIONS: Three strategies for therapeutic immunization failed to control HBV viremia in a chronically infected chimpanzee with a high viral load.     

Resolution of linear minichromosomes with hairpin ends from circular plasmids containing vaccinia virus concatemer junctions

The junctions, separating unit-length genomes in intracellular concatemeric forms of vaccinia virus DNA, are duplex copies of the hairpin loops that form the ends of mature DNA molecules present in infectious virus particles. Circular E. coli plasmids with palindromic junction fragments were replicated in vaccinia virus-infected cells and resolved into linear minichromosomes with vector DNA in the center and vaccinia virus DNA hairpins at the two ends. Resolution did not occur when the concatemer joint was less than 250 bp or when plasmids were transfected into uninfected cells, indicating requirements for a specific DNA structure and viral trans-acting factors. These studies indicate that concatemers can serve as replicative intermediates and account for the generation of flip-flop sequence variation of the hairpins at the ends of the mature vaccinia virus genome.     

Expression of beta-galactosidase in recombinant nonintegrated plasmids in evaluating the functional activity of vaccinia virus promoters

The recombinant plasmids pVL1 and pVL2 were constructed for insertion and expression of alien genetic information in HindIII-F fragment of vaccinia virus DNA under the control of the strong early-late promoter of the protein 7.5. The late promoter of the main late protein 11K of vaccinia virus was cloned. These as well as other vector plasmids have been used to express the procaryotic beta-galactosidase gene. Functional activity of the genetic engineering constructions was estimated by transitory expression of beta-galactosidase after plasmid DNA transfection into the chicken fibroblasts embryo culture infected with vaccinia virus. The promoters of the genes for 7.5K and 11K proteins permitted the high level of beta-galactosidase expression. Using of the early promoter of the central part of HindIII-F fragment DNA from vaccinia virus was less efficient for expression of the enzyme.     

Isolation of cis-acting vaccinia virus DNA fragments promoting the expression of herpes simplex virus thymidine kinase by recombinant viruses.

Recombinant TK- vaccinia viruses containing the pBR322 sequence inserted in either orientation within the coding sequence of the viral thymidine kinase gene were constructed. They were characterized by genomic analysis, hybridization studies, reversion to wild-type virus by in vivo recombination, and rescue from their genomes of plasmids which contained all or parts of the pBR322 sequence. TK- cells were infected with one of these recombinant viruses and then transfected with pools of chimeric plasmids composed of a cloned herpes simplex virus thymidine kinase gene which contained upstream inserts of different vaccinia DNA fragments prepared by restriction or sonication. Recombination between homologous pBR322 sequences within infected cells generated selectable recombinant viruses in which expression of the herpes simplex virus thymidine kinase gene was promoted by the upstream vaccinia insert. These viruses were characterized by genomic analysis, hybridization, and in vivo or in vitro phosphorylation of (5-[125I]deoxycytidine as a specific assay for the expressed herpes simplex virus thymidine kinase. Vaccinia DNA inserts were isolated conveniently for transfer to bacteria by rescuing appropriate plasmids from the genome of recombinant viruses. The sequence of 100 nucleotides adjacent to the upstream region of the herpes simplex virus gene was determined in nine different inserts measuring 0.17 to 1.07 kilobase pairs.     

Multiple-cloning-site plasmids for the rapid construction of recombinant poxviruses

Plasmid vectors containing multiple cloning sites suitable for the rapid insertion of protein-coding sequences into poxviruses have been constructed. They are based on pUC plasmids and carry the thymidine kinase (TK) gene of vaccinia virus interrupted by a vaccinia virus promoter. Six unique restriction enzyme sites (BamHI, SalI/HincII, PstI, HindIII, EcoRI), located within 40 bp of vaccinia virus promoters transposed from the HindIII-F or HindIII-C fragment of the vaccinia virus genome, allow rapid insertion of foreign-protein-coding sequences into these plasmids. Such plasmids can be used to construct recombinant poxviruses expressing foreign proteins using marker-rescue recombination techniques and selection for TK negative viruses. Vaccinia viruses expressing the haemagglutinin (HA) gene of swine influenza virus, A/NJ/11/76 (H1N1), have been constructed.     

一种以我国痘苗病毒天坛株血凝素基因为选择标记的新型系列表达载体的构建

本文作者从痘苗病毒天坛株Sal I基因文库中确定并分离了痘苗病毒血凝素(HA)基因并组建了以HA为选择标记的系列表达载体,可用于不同结构和不同要求的外源基因在真核细胞中的表达。本系列载体的优点是,在病毒重组过程中可避开传代细胞系,一次性选择出可供疫苗使用的重组病毒。而HA阴性的重组病毒的毒力在家兔皮内试验中,未见明显下降。