genetic engineering (9)

interest DNA: genomic DNA, complementary DNA chemical or enzymatic synthetic

+vector : plasmid,phage, cosmid, phagemid, BAC, YAC

=recombinant DNA-->host: bacteria, yeast, fungi, animal cells, plant cells

genetic engineering (8)

DNA modifying enzymes
T4 polynucleotide kinase-T4 Ecoli ATP at 5' or exchange reaction-5' labeling*****
Alkaline phosphatase-bacterial AP(BAP-heat satble) or calf intestinal AP (CIP-heat labile)-5' digest Phosphate group ,prevent ligase action
DNA ligase-E coli ligase (NAD) or T4DNA ligase (ATP) nick repair or link cohesive end or blunt end (T4 ligase)
Terminal deoxynucleotidyl transferase-bovine thymus-3' add dNTP without template add complementary sequence at one end of vector and one end of gene


Nuclease
NUclease Bal 31-Alteromonas espejiana Bal 31 5'->3' exonuclease , ssDNA endonuclease
Exonuclease III-E coli3'->5' exonuclease (cannot use 3' protruding)
S1 Nuclease-Aspergillus oryzae-endonuclease ssDNA, ssRNA, nick or gap in duplex DNA, or RNA use to detect non-complete complementary, produce blut end, hairpin cDNA break
Mung beab nuclease (MB nuclease)-Mung bean-ssDNA, ssRNA, gap in duplex DNA, or RNA
DNAse I- cow pancreas- endonuclease ss,ds, with Mg - random, with Mn-blunt end or 1-2 bp protruding end
RNase -E coli digest RNA hybrid with DNA