dna recombinant technology & genetic engineering (1) several concepts
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DNA recombinant technology & genetic engineering (1) several concepts * recombinant DNA * technology of recombinant DNA. * Genetic engineering general procedure A. separate B. cut C. linkage - PowerPoint PPT PresentationTRANSCRIPT
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