【生命科学知识讲解】Reombination Technology.docVIP

　 　 Recombination Technology 　 What is Recombinant DNA? What are Plasmids? What are Restriction Enzymes? How often does a Restriction Enzyme cut? What is Recombinant?DNA?? Any DNA molecule formed by joining DNA segments from different sources. What are Plasmids?? Small, circular, extrachromosomal DNA molecules. They can replicate independently of the genome, and are found in numbers ranging from one per cell to hundreds per cell (this is called copy number). Plasmids frequently carry genes for antibiotic resistance. While antibiotic resistance is becoming an increasingly important problem medically, it is a useful marker in Recombinant DNA technology. Such markers, along with the small size and potentially high copy number, make plasmids indispensible tools in Molecular Biology. The following figure is an example of a typical plasmid, called pACYC184, which is 4,240 base pairs (bp, or 4.24 kilobases, kb). This plasmid map has 3 marked regions on it. Two of them are?antibiotic resistance genes, one for Tetracycline resistance (Tc) and the other for Chloramphenicol resistance (Cm). The arrows denote the direction the genes are transcribed. Remember that transcription can only occur in the 5 -- 3 direction, but plasmids are double-stranded, so the direction needs to be explicitly stated so it is known which strand is used as template for transcription. The other marked region (ori) is the?origin of replication, the region where the DNA replication machinery assembles. What are Restriction?Enzymes?? Also known as Restriction Endonucleases, these enzymes recognize and cut specific sequences in double-stranded DNA. Discovered in bacteria in 1962, Restriction Enzymes are made to protect the bacteria from foreign DNA. Bacteria have a method of marking their own DNA as being self (called a Modification System). Any DNA not recognized as self is digested into smaller pieces by the Restriction Enzymes. Restriction Enzymes search for exact sequences of a defined length. Some e