《食品生物技术》第二章 基因克隆的酶学基础.pdfVIP

ChapterⅡThetoolsoftrade

Nucleases：

Nucleaseenzymesdegradenucleicacidsby

breakingthephosphodiesterbondthat

holdsthenucleotidestogether.

RNaseDNase

Exonuclease(核酸外切酶):

WhichdegradeDNAfromthetermini

ofthemolecule.

Endonuclease(核酸内切酶):

CutataninternalpositioninaDNA

strand

SectionIRestrictionenzymes：

WhichcutDNAatdefinedsites,

representoneofthemostimportant

groupsofenzymesforthe

manipulationofDNA.

．Restriction-modification

Wheretheyfunctionaspartofa

protectivemechanismcalledthe

restriction-modificationsystem.Inthis

systemtherestrictionenzyme

hydrolysesanyexogenousDNAthat

appearsinthecell.

λ（K）λ（B）

Topreventtheenzymeactingonthe

hostcellDNA,themodification

enzymeofthesystem(amethlase)

modifiesthehostDNAbymethylation

ofparticularbasesintherecognition

sequence,whichpreventsthe

restrictionenzymefromcuttingthe

DNA.

Thediscoveryofrestrictionenzymes

Hind

5’GTPyPuAC3’

3’CAPuPyTG5’

EcoR

5’GAATTC3’

3’CTTAAG5’

2．Endonucleaserestriction

enzymetype

Restrictionenzymesareofthree(Ⅰ,Ⅱ

orⅢ).Mostoftheenzymesusedtoday

aretypeⅡenzymes,whichhavethe

simplestmodeofaction.Inessence

theymaybethoughtofasmolecular

scissors.

三、种不同类型的限制酶具有不同的特性

Restrictionenzymenomenclature

H.O.Smith和D.Nathans（1973）

Restrictionenzymenomenclatureisbasedonanumberof

conventions.Thegenericandspecificnamesofthe

organisminwhichtheenzymeisfoundareusedtoprovide

thefirstpartofthedesignation,whichcomprisesthefirst

letterofthegenericnameandthefirsttwolettersofthe

specificname.

ThusanenzymeformastrainofEscherichia