基因工程與原理 GENE CLONING & DNA ANALYSIS T. A. BROWN 許勝傑 博士 長庚大學 生物醫學系 助理教授 生物醫學研究所 生化暨細胞分子生物學組 分子細胞與腫瘤研究室 電話 (03) #3690

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Chapter 1 Why Gene Cloning and DNA Analysis are Important 1.1 The early development of genetics 1.2 The advent of gene cloning and the polymerase chain reaction 1.3 What is gene cloning? 1.4 What is PCR? 1.5 Why gene cloning and PCR are so important Obtaining a pure sample of a gene by cloning PCR can also be used to purify a gene 1.6 How to find your way through this book

1.1 The early development of genetics Genes reside on chromosome was proposed by W. Sutton in T.H. Morgan developed gene mapping in Analysis of the relative positions of 2000 genes on the 4 chromosomes of Drosophila melanogaster. Walter Stanborough Sutton (April 5, November 10, 1916) was an American geneticist and physician whose most significant contribution to present-day biology was his theory that the Mendelian laws of inheritance could be applied to chromosomes at the cellular level of living organisms. This is now known as the Boveri-Sutton chromosome theory. Thomas Hunt Morgan (September 25, 1866 – December 4, 1945) was an American evolutionary biologist, geneticist and embryologist and science author who won the Nobel Prize in Physiology or Medicine in 1933 for discoveries relating the role the chromosome plays in heredity.

Deoxyribonucleic acid (DNA) is the genetic material ( ). In 1927 Nikolai Koltsov proposed that inherited traits would be inherited via a "giant hereditary molecule" made up of "two mirror strands that would replicate in a semi-conservative fashion using each strand as a template". In 1928, Frederick Griffith 格里夫茲 discovered that traits of the "smooth" form of Pneumococcus could be transferred to the "rough" form of the same bacteria by mixing killed "smooth" bacteria with the live "rough" form. This system provided the first clear suggestion that DNA carries genetic information—the Avery–MacLeod–McCarty experiment—when Oswald Avery, along with coworkers Colin MacLeod and Maclyn McCarty, identified DNA as the transforming principle in DNA's role in heredity was confirmed in 1952, when Alfred Hershey and Martha Chase in the Hershey–Chase experiment showed that DNA is the genetic material of the T2 phage. Streptococcus pneumoniae 肺炎鏈球菌 type II-R (rough) type III-S (smooth) Frederick Griffith

In 1953, James D. Watson and Francis Crick suggested what is now accepted as the first correct double-helix model of DNA structure. Genetic code, transcription, translation.

1.2 The advent of gene cloning and the polymerase chain reaction Recombinant DNA technology, genetic engineering, gene cloning.

Figure 1.1 The basic steps in gene cloning. 1.3 What is gene cloning?

Figure 1.2 The basic steps in the polymerase chain reaction (PCR). 1.4 What is PCR? Kary Mullis invented polymerase chain reaction (PCR) in 1985.

Figure 1.3 Cloning allows individual fragments of DNA to be purified. 1.5 Why gene cloning and PCR are so important Obtaining a pure sample of a gene by cloning PCR can also be used to purify a gene

Two limitations of PCR! The sequences of the annealing sites must be known. The length of DNA sequence.

Figure 1.4 The problem of selection.

Figure 1.5 Gene isolation by PCR.

1.6 How to find your way through this book