ENTRAPMENT OF FLUORESCENT E. COLICELLS IN ALGINATE GEL
Keywords:
fluorescent cells, alginate gell, entrapmentAbstract
By this experiment we will demonstrate the possibility to obtain genetically modified
microbial strains that can be used as markers in different studies. The trait
transferred in this study is the fluorescence in UV light expressed by a gene isolated
from jellyfish. This gene was insered into a plasmid carrying ampiciline resistance
and in the operon for arabinose fermentation. The plasmid was calledpGLO. E coli
HB101 K-12, ampicillin resistant colonies has been obtained. The colonies on the
LB/amp/ara plate fluoresce green under UV light and the transformed colonies can
grow on ampicillin. Transformation efficiency = 362 transformed colonies/ pg DNA.
The cells where immobilized by entrapment in alginate gel to study the phenomenon
involved in cells immobilization. After immobilization in alginate gel, 5x10* cells of
E. coli pGLO / capsule and 1,4 x iff cells of E. coli HBlOl/capsule has been found.
Fluorescent microscopy revealed the presence of pGLO carrying cells into the
capsules. After cultivation of alginate capsules containing E. coli in LB broth, and
fluorescent microscopy of the capsule sections, several observations of the
phenomenon involved in continuous fermentation using biocatalysts in has been
made. These cells grow and migrate to the cortical part of the matrix where they are
immobilized.
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