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NotificationsNotify me of updates to NUMBER OF PLASMIDS TRANSPORTED INTO ESCHERICHIA COLI THROUGH THE YOSHIDA EFFECT AND PREDICTED STRUCTURE OF THE PENETRATION-INTERMEDIATE, pp. 37 - 56
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NUMBER OF PLASMIDS TRANSPORTED INTO ESCHERICHIA COLI THROUGH THE YOSHIDA EFFECT AND PREDICTED STRUCTURE OF THE PENETRATION-INTERMEDIATE, pp. 37 - 56 $100.00
Authors:  Naoto Yoshida
Abstract:
When a colloidal solution containing nano-sized acicular material, such as chrysotile,
and bacterial cells are placed in friction field between hydrogel and a polymer interface,
the nano-sized acicular material penetrates bacterial cells and forms a complex called a
penetration-intermediate. This is known as the Yoshida effect. The hydrogel exposure
method is a novel technique that employs the Yoshida effect to transform prokaryotes
with a plasmid. The author of the present study has confirmed that two or more of
pUC18, pHSG298, and pHSG396 containing the same replication of origin can be
simultaneously introduced into Escherichia coli cells using the hydrogel exposure
method. Multiple plasmids were maintained stably in E. coli cells, even following cell
subculture under the selection pressure by antibiotics. Multiple plasmids were maintained
stably in E. coli cells, even after the subculture was repeated. The stability of each
plasmid in E. coli cells was as follows: pUC18, pHSG298, and pHSG396. To investigate
the number of plasmids introduced into E. coli cells through the Yoshida effect, the
author carried transformed E. coli with both pUC18 and pHSG298, which are adsorbed
onto chrysotile in a ratio of 1:1. The relative proportion of colonies obtained from a LB
plate supplemented with ampicillin, kanamycin, and both, was 6:6:1, respectively.
Subsequently, transformation of E. coli via the Yoshida effect was performed using
chrysotile adsorbed to pUC18, pHSG298, and pHSG396 in a 1:1:1 ratio. The relative
proportion of colonies obtained from a LB plate supplemented with ampicillin,
kanamycin, ch1oramphenicol, and all three combined was 15:15:15:1, respectively. It is possible that the penetration-intermediate can only incorporate one plasmid using the
Yoshida effect. pUC18- and pHSG298- bound chrysotile were prepared independently.
When E. coli cells were exposed to hydrogel using both pUC18- and pHSG298-bound
chrysotile, relative proportion of colonies obtained from a LB plate supplemented with
ampicillin, and kanamycin was 1:1, respectively. However, no colonies were obtained on
a LB plate supplemented with both ampicillin and kanamycin. This result suggests that
the structure of the penetration-intermediate with acquired plasmid DNA consists of a
bacterial cell penetrated by a single chrysotile fiber. 


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NUMBER OF PLASMIDS TRANSPORTED INTO ESCHERICHIA COLI THROUGH THE YOSHIDA EFFECT AND PREDICTED STRUCTURE OF THE PENETRATION-INTERMEDIATE, pp. 37 - 56