Recombinant DNA Technology • A set of methods used to locate, analyze, alter, study, and recombine DNA sequences • Recombinant DNA is DNA in which nucleotide sequences from two different sources (even different species) are combined in the laboratory to produce a new combination of genes
Recombinant DNA Technology • It is used to: • Probe the structure and function of genes • If I delete this gene, what happens to the mouse? • Address questions in many areas of biology • How are Bos taurus and Bos indicus related? • Create commercial products • How can I get insulin production from bacteria? • Diagnose and treat diseases • Does this child have cystic fibrosis?
Recombinant DNA Technology • Using Mendelian and other genetic analyses we have talked about, everything we have deduced about genes and DNA sequence has been indirect • With recombinant DNA technology we can isolate genes and DNA sequence, study them directly andstore it in a convenient manner that facilitates future applications • Cloning the DNA sequence accomplishes all of these
Review of steps involved in cloning • http://highered.mcgraw-hill.com/sites/0072556781/student_view0/chapter14/animation_quiz_1.html
Recombinant DNA Technology • Restriction enzymes were the key breakthrough in recombinant DNA technology and cloning • Restriction enzymes are used to cleave DNA at specific sequences • Cuts DNA at specific sequences into different sized molecules • Cuts a single phosphodiester bond on each strand
Recombinant DNA Technology • Recognition sequence • Usually four to six bases • Palindromes – read the same on both strands • Cut DNA reciprocally at specific recognition sites
Recombinant DNA Technology • Most restriction enzymes make offset cuts • Restriction sites are not opposite each other • Results in staggered cuts (sticky ends) • Ability for hydrogen bonding to a complementary sequence