ecology-ecosystem-microbial-ecology-2019-posters

16 / 18

49
conferenceseries LLC Ltd
3rd International Conference on
3rd International Conference on
Ecology, Ecosystem and Conservation Biology
Microbial Ecology & Eco Systems
&
March 18-19, 2019 | Chicago, USA
Find More Information @ MARCH 2019 Conference Series LLC Ltd
Inactivation of a
proteorhodopsin
like gene in
Aurantiochytrium by
double homologous
recombination
Kylen Bao
Stony Brook University, USA
Aurantiochytrium
limacinum, a marine
heterotroph which belongs
to a crude oil-degrading
class of protists called the
Labyrinthulomycetes, is one of
a variety of thraustochytrids
known to produce zoospores
that respond to chemical
cues, as well as light, but the
mechanisms by which they do
so are unknown. While many
papers have been published
concerning how zoospores
across different taxa respond
to light, little research has
been done in investigating
the mechanism of light
sensitivity of Aurantiochytrium
limacinum, which plays an
important role in the carbon
cycle by decomposing crude
oil, tarballs, and other
non-living organic matter.
Since rhodopsins can be
involved in phototaxis, we
hypothesized that the gene
7690 in Aurantiochytrium,
which encodes a protein
with similarity to rhodopsins,
serves as a photoreceptor
for zoospore phototaxis. By
attempting to knock out the
7690 genes through double
homologous recombination,
this research aimed to
determine whether the
7690 protein is required for
phototaxis of the zoospores.
We extracted, purified and
restriction digested a plasmid
containing the antibiotic
resistance cassette we call
‘GZG’ (made of promoter
and terminator regions of
the Aurantiochytrium GAPDH
gene surrounding, and driving
the expression of, the sh
ble gene, which encodes
resistance to zeocin) in
between DNA from upstream
and downstream of the
7690 genes, and introduced
that whole construct
into Aurantiochytrium by
electroporation. After testing 6
different isolates using colony
PCR to search for double
homologous recombinants, we
identified a singular potential
knockout which can be useful
in investigating whether the
7690 genes is required for
phototaxis.
JOURNAL OF ECOSYSTEM& ECOGRAPHY 2019, VOLUME 9 | DOI: 10.4172/2157-7625-C1-045 ACCEPTED ABSTRACTS