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fx similar to histones/ nucleosome
Special characteristic of bacteria
--> three types of DNA elements
they have multipartite genomes
chromosome: locatedin nucleoid, carries essential genes
chromid: uses plasmid part. system, carries essential genes
plasmid: uses plas. part. system. and carries noness. genes
Plasmids can be used for what?
What is the basic cloning schema
1) plasmid is cut and linearized by a specific restriction enzyme (endonuclease)
2) a fragment of foreign DNA is cut with the same enzyme
3) Plasmid and fragment are ligated together (ligase)
4) Ligation reaction is mixed with E. coli (heat shock or electroporation)
5) Some ligated plasmids will be incorporated in bacteria
6) Mixture is plated and incubated
7) Antibiotic resistance colonies with the inserted fragment are identified
Rolling circle mechanism
3' end displaces the 5' end of the newly synthesized strand: strand synthesis "rolls off" a linear copy of the circular DNA molecule
The second strand synthesis converts the linear DNA molecules to double stranded DNA. It is cut and ligated
What can conjugation enable?
mapping gene via conjugation,
Conjugation can be interrupted at various time points.
Mix for a certain time and see the genes that are transferred. The phenotype will serve as a marker.
Plate it and see which ones survive or are present.
Draw a plasmid indicating the origin of replication and the replative position of each gene
What would you like to do?
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