Site-specific recombination and transposition

Nivedita
Ph.D. (JNU-Life Sciences)
CSIR NET-JRF, GATE
nivedita229@gmail.com
Two class of genetic recombination
Conservative site-specific recombination (CSSR)
The integration of lambda genome into the chromosome of the host cell
Insertion Deletion Inversion

Three types of CSSR recombination

Structure involved in CSSR
Covalent intermediate mechanism used by serine and tyrosine recombinase
Recombination by serine recombinase
Recombination by serine recombinase
Recombination by tyrosine recombinase
Recombination by tyrosine recombinase
Mechanism of site-specific
recombination by Cre
recombinase
Mechanism of site-specific
recombination by Cre recombinase
 Transposition

•Transposition is a specific form of genetic recombination that moves certain

genetic element from one DNA site to another.

•Theses mobile genetic elements are called transposable elements or transposons.

•Transposable elements can insert in gene and disrupt its function, can modify
gene expression.

•It can used as a mutagen and DNA delivery vectors in experimental biology.
Transposition of a mobile genetic element
•Transposon elements can make up huge fraction of the genome of an organism
e. g. 50% TE in human and maize genome.

•Transposon content in different genomes is highly variable. E.g. Fly and yeast
genome are gene rich and transposon poor as comparison to human.
Types of transposable elements
•Autonomous transposon: DNA transposons that carry a pair of terminal
inverted repeats and gene encoding transposase and able to promote
transposition.

•Nonautonomous transposon: carry only the terminal inverted repeats (cis-acting

sequence) and needs helper transposon for its transposition.
Cut and paste mechanism of transposition
Cut and paste mechanism of transposition
Three mechanism for cleaving the nontransferred DNA strand
Three mechanism for cleaving the nontransferred DNA strand
Mechanism of replicative
transposition
Mechanism of
replicative transposition
Mechanism of retroviral integration and transposition of viral-like
retrotransposon
Q. Site -specific recombination results in precise DNA rearrangement, which is
limited to specific sequence. The enzymes that are important to carry out the
process are:

1. Restriction endonuclease and DNA polymerase

2. Nuclease and ligase
3. DNA polymerase and ligase
4. DNA polymerase and DNA gyrase
Q. In an experiment on transposition in an eukaryotic system, an intron was cloned
within a transposable element and allowed to transpose from a plasmid to genomic
DNA. The intron was found to be absent in the transposable element in its new
location. It is:

(1) not a case of transposition.

(2) a case of replicative mode of transposition.
(3) a case of conservative mode of transposition.
(4) a retroposon.
Q. Transposons can be primarily categorized into two types, DNA transposons and
retrotransposons. Given below is some information regarding the above.

A. Eukaryotic DNA transposons excise themselves from one place in the genome
and integrate into another site.
B. Retrotransposons are RNA sequences that are first reverse transcribed into cDNA and
then integrate into the genome.
C. Retrotransposons move by a copy and paste mechanism through an RNA intermediate.
D. As DNA transposons move via a cut and paste mechanism, there can never be an
increase in the copy number of a transposon.

Which of the statement(s) is/are true?

1. A and C
2.B and D
3. B only
4. D only
Q. Given below are some statements about prokaryotic and eukaryotic mobile
genetic elements or transposons.
1. Most mobile genetic elements in bacteria transposes via an RNA intermediate.
2. Most mobile genetic elements in bacteria are DNA.
3. Mobile genetic elements in eukaryotes are only retrotransposons.
4. Both RNA and DNA transposons are found in eukaryotes.

Choose the correct combination.

1. A and C
2. B and C
3. A and D
4. B and D
Transposition of a poly-A
retrotransposon (LINE) by
target site primed reverse
transcription
Q. Which one of the following statements about the human LINEs (Long
interspersed nuclear elements) is FALSE?

1. LINES are located primarily in euchromatic regions

2. LINEs cannot enconde all the products needed for their retrotransposition and are
dependent on SINEs for some components.
3. Of the three human LINE families LINE-1 is the only family to have activity
tranposing memebers.
4. Active LINE-1 elements processes and internal promoter located within 5’
untranslated region.
 Regulation

•Transposons control the number of copies present in the cell.

•Transposons control target site choice.

Bacterial transposon Tn10
Antisense regulation of
Tn10 expression
Transposition of Tn10 after
passage of replication fork