초록 ▼

대장균으로 부터 RecA 단백질을 순수분리하여 strand 교환반응을 검색을 검색하였다. Duplex간의 strand 교환반응에는 linear duplex DNA (32P)와 gapped circular DNA가 사용되었고, 반응결과 생성되는 nicked circular heteroduplex DNA 는 전기영동과 sutoradiography로 즉각 알아낼 수 있다. Strand 교환반응에 미치는 deletion 과 insertion의 영향을 조사해본 결과, 38염기쌍까지의 deletion 혹은 insertion 된 DNA 기질은

대장균으로 부터 RecA 단백질을 순수분리하여 strand 교환반응을 검색을 검색하였다. Duplex간의 strand 교환반응에는 linear duplex DNA (32P)와 gapped circular DNA가 사용되었고, 반응결과 생성되는 nicked circular heteroduplex DNA 는 전기영동과 sutoradiography로 즉각 알아낼 수 있다. Strand 교환반응에 미치는 deletion 과 insertion의 영향을 조사해본 결과, 38염기쌍까지의 deletion 혹은 insertion 된 DNA 기질은 RecA 에 의한 strand 교환에 별 영향이 없었으나, 120 염기쌍이 deletion(insertion)된 경우에는 strand 교환이 전혀 일어나지 않았다. 한편 DNA 기질을 alkylation 시킨후 3 strand 간의 교환반응을 검색한 결과 dimethyl sulfate 로 single stranded circle 을 처리한 겨웅 strand 교환반응이 전혀 일어나지 않았고, ethylnitrosourea 로 처리한 경우에는 strand 반응이 전혀 일어나지 않았고, ethylnitrosourea 로 처리한 경우에는 strand 교환반응이 처리기간에 따라 점점 지연됨을 관찰할 수 있었다.

Abstract ▼

RecA protein from E. coli was purified with homogeniety and used for the strand exchange reactions. The reaction between linear duplexes and circle with a single stranded gap as substrates generates nicked circular heteroduplex and linear molecules with single stranded ends. The completion of strand

RecA protein from E. coli was purified with homogeniety and used for the strand exchange reactions. The reaction between linear duplexes and circle with a single stranded gap as substrates generates nicked circular heteroduplex and linear molecules with single stranded ends. The completion of strand exchange can be demonstrated by the production of nicked circular heteroduplex DNA detected by gel electrophoresis and autoradiography using radiolabeled linear molecules. When the effect of deletion (or insertion) to the substrate DNA was tested, strand exchange was found to pass deletions(or insertions) of 38 base pairs. However, exchange was completely bloched by those of 120 base pairs .
Alkylation of the DNA substrates elicits severe effects on RecA mediated 3 stranded reactions. When single stranded circle was treated with dimethyl sulfate. strand exchanges were completely blocked and thus no nicked circular reaction products were detected. However, circular DNA ethylated by ethylnitrosourea inhibited the reactions as a function of treating time.

목차 Contents

• 1. 서 론...7
• 2. 재료 및 방법...11
• 3. 결과...21
• 4. 토의...32