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Recombinant DNA Cut and Tape Activity

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The concepts of Recombinant DNA Technology can be hard to initialy understand, especially if resourses are not available to carry out a wet lab. This printable activity gives students the opportunity to cut out components of a plasmid and artificially construct a vector with particular attributes that make it suitable to express a hypothetical gene in E. coli

Hands on activities help solidify concepts that have been introduced through text and lecture.

Recombinant DNA technology involves the cutting and recombining of DNA fragments from different sources and organisms. Fragments of DNA are typically cloned into small circular DNA fragments called plasmids which are originally isolated from bacteria. The genetically modified plasmids are then transformed into a bacterial host such as E. coli which can then transcribe and translate the genes into proteins.

Using scissors and clear tape, cut out and paste together the minimal* set of components needed to create a plasmid which can express geneX in E. coli.

Components will be needed that make it possible to: select for colonies that have the plasmid screen for colonies that have geneX cloned into the plasmid allow the plasmid to replicate in E. coli allow geneX to be induced by addition of IPTG

Plasmid components included in the printable activity include:

AmpR is a gene which can be transcribed and translated into a protein that makes bacteria resistant to the antibiotic ampicillin. It is one example of a "marker" which allows selection of bacteria that have this gene on a plasmid.

KmR is a gene which can be transcribed and translated into a protein that makes bacteria resistant to the antibiotic kanamycin.

cos represents the sequences necessary for packaging of DNA within a bacteriophage (a virus that infects bacteria).

ori represents the origin of replication sequence needed for the plasmid to be replicated in bacteria. Many different origin of replication sequences have been discovered on different plasmids.

lacZ is transcribed and translated into a protein called beta galactosidase which metabolizes lactose. It also converts X-gal (a structural analog of lactose) into a blue compound. The lac promoter (with operator) is also present to allow transcription and regulation of transcription.

lacI represses transcription of the lac operon in the absence of lactose (allolactose) or a structural analog of allolactose called IPTG.

geneX is our hypothetical gene that we want to transcribe and translate in E. coli.