Μοριακή Βιολογία Chapter 6

DNA 클로닝

• DNA 클로닝: 2개의 DNA 단편을 결합하는 공유적 방법
• 선택적 Vector: малень키 원형 DNA 분자, 클론을 위한 선택적 표지자 및 스크린 표지자 포함
• 제한 엔도핸ديد: 특정 서열에서 DNA 절단
• Expression Vector: 프로모터 포함하여 셀에서 프로틴 생산

PCR

• PCR (Polymerase Chain Reaction): 특정 DNA 서열을 증폭하는 방법
• site-directed mutagenesis: 특정 아미노산을 대체하는 방법
• restriction endonucleases: 특정 서열에서 DNA 절단
• Terminal tags: 프로틴 정제에 사용되는 태그

비유기 분석

• Green Fluorescent Protein (GFP): 생체 내에서 프로틴 분석에 사용
• Epitope tags: 특정 프로틴을 분석하는 방법
• Two-hybrid analysis: 프로틴-프로틴 교감 분석

DNA 구조

• DNA phosphodiester bonds: 3' - 5' phosphodiester 결합
• Chargaff's rules: A-T, G-C base pair rules
• Watson-Crick Model: B-DNA 구조, 두 가닥螺旋 구조
• Hoogsteen interactions: non-Watson-Crick base pairing
• Palindromic sequence: 같이 읽히는 DNA 서열

###脂質 (Lipid)

• Membrane lipids: phospholipids, glycosphingolipids, galactolipids, etc.
• Signaling lipids: prostaglandins, thromboxanes, leukotrienes, etc.
• Fatty acids: sphingosine에atty acid linked by an amide linkage
• Steroid hormones: cholesterol에서 파생된 호르몬
• Phosphatidylinositol 4,5-bisphosphate (PIP2): 신호 전달에 사용되는 리파지드

그 외

• Microarray: 유전자 expresion 분석에 사용되는 방법
• X-ray crystallography: Oswald Avery의 DNA 구조 분석 방법

phospholipids and cholesterol

• Phospholipids are present in the membranes of most eukaryotic cells, modulating fluidity and permeability.
• Cholesterol is present in the membranes of most eukaryotic cells, modulating fluidity and permeability.

Enzymatic oxidation

• Enzymatic oxidation of arachidonic acid yields prostaglandins, thromboxanes, and leukotrienes.

Steroid hormones

• Steroid hormones are oxidized derivatives of sterols.

Carbohydrates

• Aspartame is an artificial non-saccharide sweetener 200 times sweeter than sucrose.
• To possess optical activity, a compound must be asymmetric.
• The reference compound for naming D and L isomers of sugars is glyceraldehyde.

DNA structure

• The double helix of DNA in the B-form is stabilized by nonspecific base-stacking interactions via van-der-Waals interactions.
• In nucleotides and nucleic acids, syn and anti conformations relate to rotation around the sugar-base bond.
• B-form DNA in vivo is a right-handed helix, 18 Å in diameter, with a rise of 3.4 Å per base pair.

Lipids

• Triacylglycerols are composed of a glycerol backbone and three fatty acids, linked by ester bonds.
• Biological waxes are esters of single fatty acids with long-chain alcohols.
• Fatty acids are not strongly hydrophilic.
• Arachidonic acid is the precursor of prostaglandins.
• Sphingosine is a component of ceramide, cerebrosides, and gangliosides.
• Phosphatidylcholine is a glycerophospholipid, used as an emulsifier in margarine and chocolate.
• Cardiolipin is a glycerophospholipid, found in the inner membrane of mitochondria.
• Glycerophospholipids contain fatty acids linked to glycerol through ester bonds.
• Ceramide is not a glycerophospholipid.
• Sphingolipids may be charged, but are always amphipathic.

Signaling lipids

• Platelet-activating factor (PAF) is a signaling lipid produced by platelets to aid in the formation of blood clots.
• Leukotrienes are signaling lipids that cause asthmatic attacks.
• Nonsteroidal antiinflammatory drugs (NSAIDs) inhibit the synthesis of prostaglandins and thromboxanes from arachidonic acid.

Steroid hormones and other lipids

• The steroid hormones testosterone and beta-estradiol are derived from cholesterol.
• Phosphatidylinositol 4,5-bisphosphate (PIP2) serves as a reservoir of messenger molecules for signal transduction.
• Beta-carotene can be cleaved in half to generate a molecule important in vision.
• Certain lipid molecules can be used as colorful pigments in plants and animals due to their conjugated double bonds.