DNA recombinant

technology & genetic

engineering

(1) several concepts

* recombinant DNA

* technology of recombinant

DNA

* Genetic engineering

(2) general procedure

A. separate

B. cut

C. linkage

D. transfection or

transformation

E. Screening

a. separate

* Vectors

concept

type

* the sources of foreign target DNA

three methods

genomic DNA

complementary DNA,cDNA

synthetic DNA fragment

Lehninger 112601

Genomic DNA library

cDNA library

b. cut

• restriction endonuclease

• concept

• nomination

• type

5’ G-A-A-T-T-C 3’

3’ C-T-T-A-A-G 5’

5’ G 3’ 5’ A-A-T-T-C 3’

3’ C-T-T-A-A-5’ 3’ G 5’

sticky end

5’-A-G-C-T-3’

3’-T-C-G-A-5’

5’-A-G-3’ 5’-C-T-3’

3’-T-C-5’ 3’-G-A-5’

blunt end

c. Linkage

d. transformation or transfection

microinjection

transgenic

E. Screening

drug-resistance gene

X-gal screening

Southern blot

* digested with enzyme

* electrophoresis

* transfer

* hybridization

* autoradiography

Lehninger 92201