Презентация на тему: Recombinant DNA Technology

DNA,RNA, Recombinant DNA Technology

Gene expression.

X-Ray structure of a complex of ethidium bromid with DNA.

Construction of a restriction map.

Restriction map for the 5243-bp circular DNA of SV40.

Construction of a recombinant DNA molecule through the use of synthetic oligonucleotide adaptors

Insertional inactivation Gene in cloning site: LacZ -> pUC18 (lacZ complements the host defect in lacZ) -> pUC18 into host organism -> active lacZ (β-galactosidase) from plasmid-> cleavage of X-gal (blue colonies) -> gene cloned into polylinker -> lacZ gene disrupted -> no cleavage of X-gal (white colonies)

Insertional inactivation Gene in cloning site: Resistance marker -> pBR322 (cloning sites within antibiotica resistence marker) -> plasmid into host -> resistance against 2 antibiotica -> gene cloned within one resistance marker -> gene for antibiotica resistance marker disrupted -> sensitive against one antibioticum

Electron micrograph of bacteriophage λ.

Bacteriophage T2 injecting its DNA into an E. coli

Cloning of foreign DNA in λ phages.

Creation of Libraries

Creation of Libraries

Sizes of Some DNA Molecules.

Fragmentation of genomic DNA

Bacterial host engineering

E.Coli K12 strain has been used for further engineering

Additional changes in K12 E.coli for ease of the laboratory practice

Additional changes in K12 E.coli for ease of the laboratory practice

Transformation of plasmid DNA to competent E. coli cells

Calcium/phosphate (heat shock) method