DNA的生物合成概要.ppt

* tRNA as the primer * 逆转录病毒的生活史 * Reverse transcriptase was discovered by Howard Temin at the University of Wisconsin–Madison, and independently by David Baltimore in 1970 at MIT.[2][3] The two shared the 1975 Nobel Prize in Physiology or Medicine with Renato Dulbecco for their discovery. In 1986 Dulbecco was among the scientists who launched the Human Genome Project * Reverse transcriptase is used, as you might expect, to copy RNA into DNA. This task is integral to cloning complementary DNAs (cDNAs), which are DNA copies of mature messenger RNAs. Cloning cDNAs is discussed elsewhere in more depth, but the technique is usually initiated by mixing short (12-18 base) polymers of thymidine (oligo dT) with messenger RNA such that they anneal to the RNAs polyadenylate tail. Reverse transcriptase is then added and uses the oligo dT as a primer to synthesize so-called first-strand cDNA. Another common use for reverse transcriptase is to generate DNA copies of RNAs prior to amplifying that DNA by polymerase chain reaction (PCR). Reverse transcription PCR, usually called simply RTPCR, is a stupifyingly useful tool for such things as cloning cDNAs, diagnosing microbial diseases rapidly and a myriad of other applications. In most cases, standard preparations of reverse transcriptase are used for RTPCR, but mutated forms with relatively high thermal stability have been developed to facilitate the process. * an oligo dT primer is annealed to the poly A portion of the mRNA template B. the enzyme reverse transcriptase catalyzes the synthesis of DNA from an RNA template C reverse transcriptase backs around to place a hook at the end - DNA-DNA primer D. RNA. is removed by alkaline hydrolysis E. DNA synthesis is completed with DNA polymerase F. this process is used in molecular cloning * Some circular DNA molecules, including those of a number of bacterial and eukaryotic viruses, replicate by a process that does not include a theta shaped intermediate. This replication