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Introduction
Segments of the genome that are capable of moving around to different location. Transposable elements usually are flanked (Be situated on each side of or on one side of something) by repeated sequences . Carry transposase genes that confer the transposition ability . Variety of names controlling elements , jumping genes , mobile genes , mobile genetic elements , transposons. Found in phages ,viruses , prokaryotes ,eukaryotes and Insects.
Insertion Sequence
segments of bacterial DNA that can move from one position to a different position Range in size from 768 bp to 5 kb Disrupt coding sequences or regulatory regions. Alter expression of nearby genes. Cause deletions and inversions in adjacent DNA. Result in crossing-over.
Direct visualization
Wild type lambda d gal Mutated lambda d gal Electron microscopy heteroduplex,single stranded loop,no complementry sequence to wild type 800 nucleotide 4 single stranded tail
Electron micrograph of a dgal+/ dgalm DNA heteroduplex Each heteroduplex shows a single stranded buckle, or loop
Insertion sequence
7.
Small direct repeats (~5 bp) flanking the target site are created.
Transposons
Structure formed when denatured drug resistance plasmid DNA is re annealed(Reannealing is the process by which two single strands of DNA combine to form double-stranded DNA) Double stranded IR region separates a large circular loop from the Small lollipop loop Gene for drug resistance by plasmid between IR of lollypop called Transposons
Transposons
Movement of Transposons
Phage mu
a normal-appearing phage 36,000 nucleotide long can insert itself anywhere in a bacterial or plasmid genome in either orientation; mutation in the genome like IS each mature phage particle has on each end a piece of flanking DNA from its previous host no insert into genome in next generation contain its own IR sequences; but not in the chromosome ends
genetic snap fastener: phage mu can mediate the insertion of phage or drug resistant gene into a bacterial chromosome using 2 mu genome
Replicative
A new copy of the transposable element is generated in the transposition event The Transposition of Tn3 takes place through a cointegrate intermediate
Replicative transposition
Conservative Transposition
Some transposons excise from the chromosome and integrate Into the target DNA - Generation of heteroduplex and homoduplex Tn10 elements
Conservative Transposition
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Ty element
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Retrotransposition
Retroviruses
SSRNA animal viruses Reverse transcriptase E.g: mouse mammary tumor virus (MMTV) , Rous Sarcoma Virus (RSV) cancer Integrated into host chromosome as DSDNA called provirus Like mu phage considered as transposable element , can infect , transposes. Integration results in duplication of short target sequence in host chromosome
Retroviruses
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FB element
FB --- fold back Few hundreds to few thousands base pairs Long inverted terminal repeats Some time entire elements consists of inverted repeats ,central sequence separate the inverted repeats Mutation due to interruption by FB element in gene coding sequence or near a control region Excise them from the genome and promote chromosomal rearrangements at high frequencies
P-elements
Hybrid dysgenesis Drosophila Melanogaster Lab- M Cytotype female Natural P- Cytotype male Sterility, high mutation rate ,high frequency of chromosomal aberation and non disjunction Result dysgenic or biologically defected Vice versa no mutation Large no of mutations are unstable---wild type
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Eye color locus Insertion of genetic element in to the middle of the white gene Called P-element 30-50 copies per genome in P-strain 0.5 -2.9 kb 31 bp perfect inverted repeats at their ends
P-Element
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He first hypothesized that two events had occurred at unlinked loci: 1. Pigment gene A1 had mutated to colorless mutant a1 2. At another locus, a dominant allele for dotting (Dt) had appeared What was causing the dotted phenotype? - Reverse mutation of a A some are completely pigmented type - a : unstable mutant allele (high reversion rate) - the allelic instability is dependent on the unlinked Dt gene
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