Electrophoretic analysis of soluble proteins specifically synthesized under phosphate deficiency in the mycelia ofPholiota nameko

Mycelial soluble proteins ofPholiota nameko labeled in vivo during the Pi-supplied (P⁺) and the Pi-depleted (P⁻) cultures were separated by SDS-polyacrylamide gel electrophoresis and two-dimensional polyacrylamide gel electrophoresis, and visualized by fluorography. A comparison of protein profiles from the P⁺ and P⁻ cultures showed that Pi deficiency induces the synthesis of 15 polypeptides and an increase in the relative amount of 29 polypeptides. These result suggests that many proteins may be specifically synthesized de novo under Pi deficiency as part of the adaptive mechanism for this condition.     

Hemolytic Activity of a Togavirus,Getah*

A purified toga-alphavirus, Getah (GET), showed optimal hemolytic activity for one-day-old chick red blood cells when incubated at 37 C for 120 min at pH 6.2. Experimental data obtained from various angles, such as pH dependency, inhibition by virus-specific antiserum and by concanavalin A, indicated that the hemolysis was a property of the virus particle itself. Although the mechanism of hemolysis by togaviruses has not been known, our results indicated that viral lipids may participate in this activity since the hemolytic activity was impaired by delipidation procedures.     

Automated in vitro selection to obtain functional oligonucleotides.

In vitro selection or systematic evolution of ligand by exponential enrichment (SELEX) has been devised for the identification of high-affinity oligonucleotide aptamers to target molecules. However, the selection process is repetitive and time-consuming. We have developed an automatic for in vitro selection by assembling an affinity chromato-column, a PCR thermal cycler, a HPLC and a sample operation system. Several molecular biology methods were optimized for the machine. Automated selection was used to generate nucleic acid aptamers interacting specifically with an environmental contaminant.