un seminario hecho con mucho esmero y dedicacion

1. ANDREAPALACIO MONTOYA
TERCER SEMESTRE
MEDICINA
UNIVERSIDAD PONTIFICIA BOLIVARIANA

2. INTRODUCTION
LactobacillusReuteriN16
L. reuteri, which was first isolated in 1962, has been recognized as an
obligatory heterofermentative and endogenous lactic acid bacterial
species in the GI tract of mammals. Furthermore, specific L. reuteri
strains have been found to display various beneficial effects, including
the production of antimicrobial molecules such as organic acids,
ethanol, and reuterin . Several L. reuteri strains can decrease the levels
of pro-infammatory cytokines and thus promote the function and
development of regulatory T cells. In addition, specifc L. reuteri strains
have been shown to reduce the incidence and severity of diarrhea,
prevent colic and necrotic enterocolitis, and maintain a functional
mucosal barrier in humans.

3. PlasmidpTE15
Plasmids are circular or linear extra-chromosomal DNA molecules that
replicate and transcribe independently of chromosomal DNA. They are
normally present in bacteria, and sometimes in eukaryotic organisms
such as yeast.
Plasmid DNA molecules adopt a double helix conformation like the
DNA of chromosomes, although, by definition, they are outside of
them. Plasmids have been found in almost all bacteria. Unlike DNA,
chromosomes, plasmids do not have associated proteins.
INTRODUCTION

4. GENERALOBJECTIVE

5. METHODS
 SEQUENCING
It is an experiment in which the sequence of nucleotides that
make up a strand of DNA is determined.
When analyzing the sequence of the lactobacillus plasmid
and seeking to know how it was genetically made. Universal
primer was used according to the procedures described in
the Sequenase 2.0 enzyme kit (US Biochem). Analysis of DNA
sequence with gene finding and pattern recognition was
performed using Blastx and GCG Wisconsin Package
(UWGCG) provided by the Genetics Computer Group.
 ELECTROFORESIS
Electrophoresis is a laboratory technique used to separate
DNA, RNA, or protein molecules based on their size and
electrical charge. An electrical current is used to move the
molecules through a gel or other matrix. The pores in the gel or
matrix act like a sieve, allowing smaller molecules to move
faster than larger molecules.
 Molecular manipulations, such as genome and plasmid
isolation, electrophoresis, restriction endonuclease digestion,
and fragment ligation, were performed according to standard
techniques

6. METHODS
 PLASMID CONSTRUCTION
The plasmid is introduced into bacteria through a process called
transformation, and bacteria containing the plasmid are selected for
by antibiotics. Bacteria with the correct plasmid are used to make
more plasmid DNA or, in some cases, gene expression is induced to
make protein.
. To further confirm which fragments may contain the needed
regions of replication origins, pTE15-RO was digested and subcloned
as described
 RESTRICTION
Restriction enzymes are used to cut the DNA strands of two
plasmids, producing DNA fragments with complementary sticky
ends that can be reassembled to create a recombinant plasmid.

7. RESULTS
In this image you can focus on lines 5
and 6 this shows us the presence of
DNA so the bands of cars 5 and 6
contain replicated DNA derived from
L. reuteri and that grew in the medium
with the MRS compounds which is why
they both look like pounds that are
parallel.

8. RESULTS
In this other one we are going to
concentrate on the image on the
left, the A that has the most
significant findings, and they are that
the ssDNA grows with or without
rifampicin, that is to say that the
bacteria from which they took the
RNA has mechanisms of resistance to
rifampicin and that is why it grows in
the lines that it has + and that it has -
and the other lane where there is
only one line corresponds to the
location of the ssDNA, that is, how
much it weighs

9. DISCUSSION
AUTOR PROPUESTA COINCIDE
Wu Y-C These plasmids rely exclusively on host factors
for both double-strand melting and primer
synthesis; thus, their replication does not
depend on plasmid-encoding Rep proteins.
YES
Takechi S, Matsui H,
Itoh T
The Rep protein in class C plasmids displays
primase activity, synthesizing a unique primer
RNA (ppApGpA) that is extended by DNA
polymerase I at a fxed site in the origin región.
YES
Weaver KE, Clewell
D, An F
When compared RepB and RepC of pTE15 with
RepB and RepC of pAD1 using Blastp, which
previous had been functional study, the
similarity for amino acids was approximately 57
and 32%, respectively. Te RepB involved in
plasmid copy control and RepC may be
involved in stable inheritance.
YES

10. CONCLUSION
➢ Gracias a las diferentes técnicas utilizadas en este articulo, pudimos observar las
diferentes características del plásmido pTE15 de L reuteri N16 y se pudo clasificar como
una clase de tipo theta diferente en los lactobacilos
➢ Es muy importante seguir desarrollando técnicas de laboratorio enfocadas en la biología
molecular para continuar con el estudio de moléculas y microorganismos beneficios y
patológicos para el cuerpo humano.

11. MAPACONCEPTUAL
BACILOS
Se dividen en
Bacterias que al microscopio se
observan en forma de bastón
Gram negativas
Gram positivas
Pueden ser
Patógenas Benéficas
Tinción purpura o
azulada
como
Bacterias lácticas Excretan vitaminas
y prebióticos
Mejoran el desarrollo de la
microbiota del tracto GI y
pueden tener efectos
inmunomoduladores
Tinción rosa o roja
Posee una pared
celular gruesa
Evita que los
glóbulos rojos las
ingieran
Pueden ser
Patógenas Benéficas
Suelen ser muy
resistentes a los
antibióticos

12. PLÁSMIDO
Microorganismos microscópicos
Bacterias
Los científicos usan métodos de ADN
recombinante para insertar el
plásmido en el gen que se desea
estudiar
Están separados físicamente
del ADN cromosómico
Cuando el plásmido se copia a
si mismo, también copia el gen
insertado
Se pueden trasmitir de
célula a célula
Tienen un numero reducido de
genes (algunos son asociados a
la resistencia contra antibióticos
Molécula pequeña
de ADN circular
Se encuentra presente en
Se replican de
manera
independiente
Los círculos que tienen el
ADN clonado se llaman
PLÁSMIDOS
Los PLÁSMIDOS son una
herramienta fundamental de la
tecnología del ADN
recombinante