Manipulating photosynthesis

The levels of individual photosynthetic proteins can be independently decreased by theAgrobacterium-mediated transformation of plants with antisens RNA constructs. Protocols for the introduction of such constructs intoAgrobacterium, theAgrobacterium-mediated transformation of tobacco leaf disks, and the screening and analysis of the transgenic plants produced are described.     

有机盐对基因启动子的σ因子选择性的影响

用σ￣（７０）或σ￣（３８）和核心酶（Ｅ）组成的大肠杆菌ＲＮＡ聚合酶全酶（Ｅσ）对四种启动子进行了体外转录。结果表明：ｌａｃＵＶ５,ｒｐｌＪ主要被Ｅσ￣（７０）识别,ｋａｔＥ主要被Ｅσ￣（３８）识别,ｆｉｃ在低盐浓度时被Ｅσ￣（７０）识别,在高浓度盐时被Ｅσ￣（３８）识别;Ｅσ￣（３８）转录特异性启动子所需酶量大于Ｅσ￣（７０）转录特异性启动子的酶量;在含有分别对σ￣（７０）或σ￣（３８）亲和性大的混和启动子的体外转录中,启动子之间不存在干扰和竞争,转录水平与启动子浓度成正相关。     

Enzymatic labeling of nucleic acids

Enzymatic labeling of nucleic acids is a fundamental tool in molecular biology with virtually every aspect of nucleic acid hybridization technique involving the use of labeled probes. Different methods for enzymatic labeling of DNA, RNA and oligonucleotide probes are available today. In this review, we will describe both radioactive and nonradioactive labeling methods, yet the choice of system for labeling the probe depends on the application under study.     

Diazepam increases membrane fluidity of rat hippocampus synaptosomes

Diazepam in vitro produced a concentration-dependent increase of membrane fluidity in crude synaptic membranes from rat hippocampus, but not cerebellum. Similar effects were obtained with higher concentrations of Ro 15-1788 and PK 11195, while zopiclone was completely inactive. In vivo acute treatment with diazepam and Ro 15-1788 gave results similar to those in vitro. The specific benzodiazepine antagonist also significantly increased membrane fluidity and was not able to reverse diazepam's effect. The data are discussed in terms of a possible role of protein kinase inhibition by the drugs not mediated by the 'central' or 'peripheral' type of benzodiazepine receptors.     

Effect of rye embryo ribosome nuclease on double-stranded RNA

In extracts obtained by treating rye embryo ribosomes with 0.5 M NH₄Cl, nuclease activity was noted towards double-stranded RNA from virus of Penicillium chrysogenum and towards synthetic poly (A)-poly (U) and poly (I)-poly (C) complexes.     

Cell free synthesis of some storage protein subunits by polyribosomes and RNA isolated from developing seeds of pea (Pisum sativum L.)

Polyribosomes which have template activity in the wheat germ system have been isolated from developing pea seeds. Some of the translation products have identical mobilities to the vicilin and legumin subunits by SDS-PAGE. Certain products were specifically immunoprecipitated with antisera prepared against purified vicilin and legumin fractions. Various RNA fractions including poly A-rich RNA have also been isolated from polyribosomes and shown to direct the synthesis of polyripeptides whose properties are similar to the storage protein subunits. The results are discussed in relationship to other investigations with seed storage protein biosynthesis in vitro.Abbreviations DTT dithiothreitol - SDS-PAGE SDS-polyacrylamide gel electrophoresis - TCA tricarboxylic acid     

Early metabolic changes in regenerating microfragments of the liverwort Riella helicophylla (Bory et Mont.) Mont.: Protein and RNA synthesis,free α-amino concentration

Microfragments of constant size were isolated from the thallus of Riella by a rapid punching procedure. Thus it was possible to study various metabolic parameters especially during the first hours after fragment isolation. Protein synthesis increases rapidly after a slight decrease at 30 min. The earliest significant increase in RNA synthesis occurs at 8 h, indicating a different activation pattern. The concentration of -amino compounds drops at 30 min and then increases continuously, thus exhibiting a close relationship with the measured alterations in protein synthesis. Another indication of metabolic conversions during regeneration is provided by the changing level of free radioactive leucine, which shows a marked turning point at 24 h after fragmentation. Analysis of free -amino concentrations in small regions of larger fragments also indicates the establishment of new intercellular correlations only a few hours after isolation of the cells from the meristem. Up to the 8th h after fragmentation, the contents of both the adaxial and peripheral fragment regions increase. Thereafter only the adaxial (regenerating) cells continue accumulating; the peripheral (nonregenerating) ones remain at the same level.     

Kinetics of rapid RNA evolution in vitro

Summary A rapidly acquired partial resistance to the replicase antagonist, ethidium bromide (EB), seen by Spiegelman and coresearchers in Q RNA variants competitively replicating under defined conditions in vitro, reflected existence of a pool of mutant RNA molecules, preadapted to EB, and their cross-propagation from the pre-EB optimum species, MDV-1, and from other kindred variants, some of which remained undetected, according to this quantitative analysis of midivariant RNA replication kinetics. DNAlike features of their evolution, such as the cloning of variants from an MDV-1 subtype and a complicance with the fundamental theorem of natural selection, resulted from the suppression, both real and apparent, of intrinsic RNA heterogeneity through sampling and detection methods, and also by the ascendency of self-propagation over cross-propagation with advancement of a superior variant. The deficit in mean polymer fitness, compared with optimum levels, determines the lower limit of this heterogeneity. Stability conditions for frequency equilibrium and strategies for counteracting viral drug resistance have been considered.