Transposition

Presented by: Pradeep Kumar CIFE- Mumbai.

Introduction
‡ Transoposon are DNA element that can hop or transpose from one place to another and first discovered by Barbra Mc Clinteck in 1950 in corn and about 20 year later in bacteria. ‡ It exist in all organism on earth e.g in human almost half of an DNA may be transposons.

Transposition
‡ Movement by a transposon is called transposition and carried out by an enzyme called transposases and it is encoded by transposons itself so that it is called jumping gene. In one sentence:- the net result of transposition is that the transposon appears at a place in DNA different from where it was originally but all DNA movenment is not called transposition such as homing DNA element & Retrotransposons

Role of transposon
‡ Offer a way of introducing genes from one bacterium into the chromosome of anopther bacterium to which it has little DNA sequence homology.

Way of transferring transposons
‡ Can be cut out of one DNA and then inserted into another DNA or can be copied and then inserted elsewhere. It is achieved by transposase enzyme that cuts the donar DNA at the ends of transposons and then insert the transposons into the target DNA. ‡ It is highly regulated and occur only rarerly ‡ Frequency of transposition :- about once in every 10³ cell division to about once in every 10 cell division.

Structure of bacterial transposon
‡ Many different type of bact. Transposons are there , some are smaller of about 1000 bp long and carry only the genes for transposases that prometer the movement in DNA where as larger transposon may contain one or more other genes such as those which is resistant to Ab. ‡ All bact. Transposons contains repeats at their end, which are usually inverted repeat(fig 9.2) and also presence of short direct repeats of target DNA that bracket the transposons & during insertion of transposons , this sequence duplicated.

Types of bact. transposons
1. Insertion seqence element:- smallest transposons having 750 to 2000 bp length and encode little more than transposase enzyme that promote their transposition . They didn t carry selectable genes and discovered only because they inactivate a gene if they happen to hop into it and causing a detectable phenotype change. There are around 700 IS element found in bact. And in plasmid also found which are imp. for formation of Hfr strains.

Conti
2. Composite transposons :- when two transposons of the same types form a larger transposons called a composite transposons, by brackating other genes.

Types of transposition
1. Outside End transposition :- this is for transposition of a composite transposons . Here each IS element can transpose independently as the transposase acts on both of its end.

Conti..
‡ Inside end transposition:- the transposase encoded by one IS element in a composite transposons can also act on the inside end of both IS element i.e. two ends that are closest to each other. After inside end transposition there may be chance of creation of a new composite transposons .

Conti.
3. Non composite transposition

How to assay transposition
1. By observing phenotypic expression 2. Suicide vector

3. Mating out assay for transposition

Molecular model for transposition
1. Replicative

Replicative transposition

1. transposase

Note: the cuts are on different strands

Replicative transposition

2. Ligation to target ends

Replicative transposition

3. 3·-ends prime replication

Replicative transposition

4. Formation of co-integrate and resolution (resolvase TnpR)

2. Cut and paste or conservative mechanism

Relationship between replicative and Cut and paste transposition

Target site specificity

Regulation of transposition

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