Construction of targeting vectors to disruptPAT1 gene inArabidopsis Trp1-100

To develop the gene targeting system by homologous recombination inArabidopsis thaliana, we constructed two targeting vectors and showed the reliability of the scheme which is based on genetic complementation of phosphoribosylanthranilate transferase (PAT1) gene. ThePAT1 gene, which is essential for tryptophan biosynthesis, was selected as a target gene because the loss of function leads to fluorescence phenotype due to the accumulation of anthranilic acid derivatives. pHS113 containsPAT1 gene surrounding 5′ and 3′ flanking portions, but the most coding region of thePAT1 gene is replaced by the neomycin phosphotransferase gene (NPTII). pHS117 consists of 1.1 kb internal fragment of genomicPAT1 gene following withNPTII gene. In this targeting strategy,Arabidopsis PAT1 gene can be disrupted by single-step of transformation experiment.     

Microplate well coverage mixing using superhydrophobic contact

Two important challenges in microplate instrumentation are to achieve full well sample coverage and complete mixing. An effective approach of using superhydrophobic rods to accomplish these challenges is reported here. Experiments conducted showed that analytes above 50μl could be made to completely cover the bottom of 96-well standard and transparency microplates. Complete mixing was accomplished by moving the rod parallel to the well bottom while contacting the liquid. The approach is simple and controlled, and it minimizes the problems of spillage and cross-contamination. It works with analytes with varied volumes and of different viscosities present in each well of the microplate.     

Visual space distortion

We are surrounded by surfaces that we perceive by visual means. Understanding the basic principles behind this perceptual process is a central theme in visual psychology, psychophysics, and computational vision. In many of the computational models employed in the past, it has been assumed that a metric representation of physical space can be derived by visual means. Psychophysical experiments, as well as computational considerations, can convince us that the perception of space and shape has a much more complicated nature, and that only a distorted version of actual, physical space can be computed. This paper develops a computational geometric model that explains why such distortion might take place. The basic idea is that, both in stereo and motion, we perceive the world from multiple views. Given the rigid transformation between the views and the properties of the image correspondence, the depth of the scene can be obtained. Even a slight error in the rigid transformation parameters causes distortion of the computed depth of the scene. The unified framework introduced here describes this distortion in computational terms. We characterize the space of distortions by its level sets, that is, we characterize the systematic distortion via a family of iso-distortion surfaces which describes the locus over which depths are distorted by some multiplicative factor. Given that humans' estimation of egomotion or estimation of the extrinsic parameters of the stereo apparatus is likely to be imprecise, the framework is used to explain a number of psychophysical experiments on the perception of depth from motion or stereo. Received: 9 January 1997 / Accepted in revised form: 8 July 1997     

Endogenous level of 28-Norcastasterone is strictly regulated in Plant Cells

A cell-free enzyme solution prepared from cultured cells ofPhaseolus vulgaris mediated C-24 methylation of 28-nor-castasterone to castasterone with the aid of S-adenosylmethionine as a co-substrate in the presence of the NADPH cofactor. This enzyme solution also catalyzed conversion of 28-norcastasterone to a demethylated 28-norcastasterone, most likely 26,28-didemethyl-castasterone, when S-adenosylmethionine was not added to the enzyme solution. Furthermore, gene expression ofArabidopsis CYP85A1 andCYP85A2 mediating the conversion of 6-deoxo-28-norcastast-erone to 28-norcastasterone was strongly inhibited by treatment of 28-norcastasterone. These results suggest that 28-norcastasterone, along with castasterone and brassinolide, is an important brassinosteroid whose endogenous level should be strictly controlled to express brassinosteroid activities in plants.