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• “Transposable elements” • “Jumping genes” • Mobile DNA
– able to move from one place to another within a cell’s genome – sometimes a copy is made and the copy moves – insertion requires target DNA sequences
inverted terminal repeat (ITR)
.• In the process. they may . .induce chromosome breakage and rearrangement.increase (or decrease) the amount of DNA in the genome.cause mutations.promote genome rearrangements. . .regulate gene expression. .
Discovery of transposons • Barbara McClintock 1950’s Ac Ds system in maize influencing kernel color unstable elements changing map position promote chromosomal breaks. • Rediscovery of bacterial insertion sequences source of polar mutations discrete change in physical length of DNA inverted repeat ends: form “lollipops” in EM after denaturation. .
use reverse transcriptase to make a DNA copy of the RNA to insert in a new location .These mobile segments of DNA are sometimes called "jumping genes" There are two distinct types of transposons: 1) DNA transposons -transposons consisting only of DNA that moves directly from place to place 2) Retrotransposons .first transcribe the DNA into RNA and then .
Classification of Transposons into two classes In both cases ds DNA intermediate is integrated into the target site in DNA to complete movement .
they are known as insertion sequences. . ● Transposons in bacteria usually carry an additional gene for function other than transposition---often for antibiotic resistance. ● Bacterial transposons of this type belong to the Tn family. When the transposable elements lack additional genes.BACTERIAL TRANSPOSONS ● In bacteria. transposons can jump from chromosomal DNA to plasmid DNA and back.
Insertion sequence 2.Tn3-type transposon 4.BACTERIAL TRANSPOSONS .Transposable phage .Composite transposon 3.TYPES 1.
. coli DNA. are examples of DNA transposons. coli genome may contain 20 of them.Insertion sequences Insertion sequences – IS1 and IS186.1. IS elements transpose either replicatively or conservatively. present in the 50kb segment of the E. Single E. Most of the sequence is taken by one or two genes for transposase enzyme that catalyses transposition.
Higher rates result in greater mutation rates.molecular parasites in bacterial cells. IS elements Study of E. IS . Transposition of IS is very rare – one in 105-107 cells per generation.cont…. coli mutations resulting from insertion of 1-2 kb long sequence in the middle of certain genes. Inserted stretches or insertion sequences – could be visualized by EM. .
The length of these repeats is constant for a given IS element. The 5’ and 3’ short direct repeats are generated from the target-site DNA during the insertion of mobile element. . but their sequence depends upon the site of insertion and is not characteristic for the IS element. It is flanked by inverted repeats of characteristic sequence.Bacterial IS element Central region encodes for one or two enzymes required for transposition. Arrows indicate orientation.
of.copies/ genome 8 5 1 or more 1 or more .Insertion sequences in E.coli Elements IS1 IS2 IS3 IS4 Size (bp) 768 1327 1300 1426 No.
are basically the pair of IS elements flanking a segment of DNA usually containing one or more genes. They use conservative method of transposition. often coding for AB resistance.2. .Composite transposons Bacteria contain composite mobile genetic elements that are larger than IS elements and contain one or more protein-coding genes in addition to those required for transposition: Composite transposons .
Tn10 .Composite transposon .Cont… 2. Tn5.Terminal IS can transpose by in self Ex. Tn9.Flank by IS element (inverted or directed repeat) .Antibiotic resistant gene .
code for Transposase.3.Function of resolvase Decrease Transposase production Catalyse the recombination of transposon . β-lactamase.5000 bp . Resolvase . Tn 3 transposon family .
lactamase – Resistance gene .Cont… ITR ITR transposase resolvase β-lactamase • Tn3 – type transposon --.inverted terminal repeat • β.5kb • ITR .
Transposable phage • Transposable phages – bacterial viruses which tranpose replicatively as a part of their normal infectious cycle. • Integrate into E. coli chromosome at regulatory element • Eg. Mu phage .4.
Transposable phage ITR ITR Integration and Replication genes Lysis genes Protein coat genes • Transposable phage – 38kb • ITR .inverted terminal repeats .
Transposable phage .Mu phage .
resulting in copying of the donor element Conservative transposition – involving excision of the element and reintegration at a new site. .Mechanism of transposition Two distinct mechanisms of transposition: Replicative transposition – direct interaction between the donor transposon and the target site.
Replicative transposition Copy of transposon sequence Transposase enzyme cut target DNA Transposition Duplication of target sequence .Mechanism of transposition 1.
Replicative transposition .
IS10.Transposition by cut and paste model Cut transposon sequence from donor molecule attach to target site Ex.2. Tn10 .Cannot copy transposon sequence . Non-replicative (conservative) transposition .
Non-replicative (conservative) transposition .
Mechanism of transposition .
• Duplications and DNA rearrangements contributed greatly to the evolution of new genes. . • It arisen once and then spread to other kingdoms by horizontal gene transfer.Evolution of Transposons • Transposons are found in all major branches of life.
. . • The evolution of an enormous variety of antibiotic resistance transposons and their spread among bacterial species.g. the fibronectin gene).Cont… • Mobile DNA most likely also influenced the evolution of genes that contain multiple copies of similar exons encoding similar protein domains (e. example of genetic adaptation via natural selection.
They can damage the genome of their host cell in different ways: 1. 2. A transposon or a retroposon that inserts itself into a functional gene will most likely disable that gene.Multiple copies of the same sequence.After a transposon leaves a gene. one of the main reasons for chromosome duplication. .Transposons causing diseases • Transposons are mutagens. the resulting gap will probably not be repaired correctly. such as Alu sequences can hinder precise chromosomal pairing during mitosis and meiosis. resulting in unequal crossovers. 3.
Cont… • Diseases caused by transposons include -hemophilia A and B -severe combined immunodeficiency -Porphyria -Cancer -Duchenne muscular dystrophy .
the first transposon to be molecularly isolated was from a plant (Snapdragon). • Researchers use transposons as a means of mutagenesis. . corn species). • Transposons have been an especially useful tool in plant molecular biology.Applications • The first transposon was discovered in the plant maize (Zea mays. • Likewise. and is named dissociator (Ds).
• To study gene expression. . • Transposons are also a widely used tool for mutagenesis of most experimentally tractable organisms. • To study the chemical mutagenesis methods.Cont… • To identifying the mutant allele.
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