PL4: DNA cloning and experimentation

vector

fragment of DNA w/ replication origins, restriction enzyme sit, and selectable marker

plasmid

most common vector used in recombinant DNA technologies that is circular, double stranded DNA, extrachromasomal, with replication occurring before cell division; 3 key attributes are 1. replication origin 2. polylinker 3. gene conferring drug resistance

extrachromasomal

existing separately from the bacterial chromosome

polylinker

artificial sequence w/ sites for many restriction enzymes adjacent to each other, allowing scientists to control how DNA goes in

transformation

changing cells from DNA outside of them, aided by CaCl2 and heat; process can occur in bacteria in nature

restriction endonucleases

enzyme that cleaves phosphodiester bonds, usually symmetrically, which can recognize a particular sequence in DNA, bind to it, then cut it

sticky end

piece of DNA cleaved by restriction endonucleases that can base pair together

BamHI

protein that makes sticky ends to cut vector to make DNA libraries, by recognizing a 6 pair base site

Sau3A

protein that recognizes a 4 base recognition site of the inner 4/6 bases that BamHI recognizes, producing same sticky ends at BamHI

reverse transcription

RNA being transcribed into complementary DNA, using reverse transcriptase

genome

the entirety of an organism’s hereditary information