生物資訊的演算法.pptVIP

1- Chapter 5 5-1 Introduction – Gene History 1865 Mendel: The basic unit of inheritance is a gene. Mendel’s work was forgotten until 1900s. 1944 The gene was known to be made of DNA (Deoxyribonucleic Acid). 1953 James Watson and Francis Crick : Double helical structure of DNA. 動物細胞 DNA位於細胞核內之「核仁」 5-2 DNA 的構造 DNA的成分與化學鍵結： 核甘酸 核甘酸(Nucleotide)為核酸分子構成單元 核甘酸包含： 五碳糖、磷酸基、含氮鹼基 核甘酸構成 DNA and RNA Nucleotide (核甘酸)的含氮鹼基有五種： adenine、guanine、cytosine、thymine、uracil DNA(deoxyribonucleic acid , 去氧核糖核酸) {A, G, C, } RNA(ribonucleic acid, 核糖核酸) {A, G, C, } DNA四種含氮鹼基 DNA Length The total length of the human DNA is about 3?109 (30億) base pairs. 1% ~ 1.5% of DNA sequence is useful. # of human genes: 30,000~40,000 Genes: 5-3 DNA 定序 5-3-1 DNA 剪斷與連接 Given DNA sequence: TGCACTTGAACGCATGCT Cut the sequence after random A: ATGCT length= ACGCATGCT length= AACGCATGCT length= ACTTGAACGCATGCT length= 5-3-2 電泳分離法 5-3-3 DNA 定序法 5-4 基因工程5-4-1 基因重組的應用 5-4-2 定點突變 5-4-3 RFLP 限制脢圖譜多形性 * * Molecular Biology-DNA Double Helix Base Pairs： G≡C、A＝T Pharmacia (1) 打破細胞 (2) 加入乙醇 (3) 以玻棒撈取 DNA (4) 乾燥後溶出 DNA (5) 以限制脢切開 DNA (6) 以電泳檢視 Restriction Enzyme GAATTC GAATTC G AATTC G AATTC Sticky Ends (Cohesive Ends) EcoRI CIVIC, Madam Get An Apple To The Class G AATTC G AATTC Arber, Nathans, Smith (1978) Juang RH (2004) BCbasics 以限制脢及連結脢進行核酸剪接 GAATTC CTTAAG GAATTC CTTAAG G CTTAA AATTC G AATTC G G CTTAA G CTTAA AATTC G G CTTAA AATTC G G CTTAA AATTC G EcoRI DNA Ligase EcoRI sticky end EcoRI sticky end Juang RH (2004) BCbasics 核酸的 Hairpin 或 Cruciform 構造 Nelson Cox (2000) Lehninger Principles of Biochemistry (3e) p.339 Sangers Method: Maxam-Gilberts Method: 做分離鹼基電泳的方法 32P 32P ATCGATCG 32P ATCG AT ATCGAT ATCG TAGCTAGCTA TAGCTAGCTA TAGCTAGCTA ATCGA 32P Specific Reaction to G ATCG A 生合成法 化學法 Template or 32P A,T,C,G A An