We aim to genetically modify unicellular microalga Chlamydomonas reinhardtii by intro
ducing
nfsI gene of bacterium Enterobacter cloacae in order to
investigate how nitroreductase expressing-microalgae respond to trinitrotol
uene
(TNT) exposure. Our experimental design is as follows: obtain a synthetic g
ene
of nfsI with flanking prefix and
suffix of standard Biobricks, and ligate this insert to pRbcBRL, a vector w
ith
appropriate expression and selection system for Chlamydomonas reinhardtii and obtain pRbcnfsI. Then, we can
transform Chlamydomonas reinhardtii
with designed plasmid. After engineering microalgae, we will grow them in t
he
presence of TNT and investigate effectiveness of nitroreductase activity on
biological degradation of TNT.