DENATURATION BEHAVIOR OF DNA-PROTEIN-COMPLEXES DETECTED INSITU IN METAPHASE CHROMOSOMES IN SUSPENSION BY HOECHST 33258 FLUORESCENCE

被引：5

作者：

SCHALLER, RKA ^{[1
]}

SPIESS, RB ^{[1
]}

BIER, FF ^{[1
]}

BETTAG, U ^{[1
]}

CREMER, C ^{[1
]}

机构：

[1] UNIV HEIDELBERG,INST APPL PHYS 1,ALBERT UBERLE STR 3-5,W-6900 HEIDELBERG,GERMANY

来源：

BIOPHYSICAL CHEMISTRY | 1990年 / 38卷 / 1-2期

关键词：

Denaturation behavior; DNA denaturation; DNA-protein complex; Helix-coil transition; Hoechst; 33258; fluorescence; Metaphase chromosome;

D O I：

10.1016/0301-4622(90)80040-E

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The denaturation behaviour of DNA-protein complexes in metaphase chromosomes in suspension was analysed in situ by Hoechst 33258 fluorescence. The results indicate that due to the stability of the dye molecule and the product of the molecular extinction coefficient and the quantum yield at different temperatures, Hoechst 33258 is a suitable probe for the detection of double-stranded DNA. Thus, it is possible to monitor the concentration of double-stranded DNA in a suspension by measuring the total fluorescence intensity. The fluorescence denaturation profiles of DNA (calf thymus) were found to be comparable to absorption measurements. The decrease in fluorescence of metaphase chromosomes in suspension with increasing temperature may therefore be used to detect conformational changes of DNA in situ. © 1990.

引用

页码：59 / 65

页数：7

共 27 条

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