RAPID ISOLATION OF METAPHASE CHROMOSOMES CONTAINING HIGH MOLECULAR-WEIGHT DNA

被引：92

作者：

BLUMENTHAL, AB ^{[1
]}

DIEDEN, JD ^{[1
]}

KAPP, LN ^{[1
]}

SEDAT, JW ^{[1
]}

机构：

[1] UNIV CALIF SAN FRANCISCO, DEPT BIOCHEM & BIOPHYS, SAN FRANCISCO, CA 94143 USA

来源：

JOURNAL OF CELL BIOLOGY | 1979年 / 81卷 / 01期

关键词：

D O I：

10.1083/jcb.81.1.255

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Metaphase chromosomes with high molecular weight DNA were isolated from Chinese hamster ovary (CHO) cells in a neutral buffer containing polyamines and chelators. The individual, unfixed chromosomes retained their centromeric and secondary constrictions, distinct sister chromatids, and complex banding patterns. The DNA from these chromosomes was 100-fold larger (2x108 daltons) than DNA from chromosomes isolated by other procedures. These characteristics indicate preservation during isolation of considerable native structure. In contrast to chromosomes produced by other methods, these chromosomes were stable in storage and did not aggregate, thus providing useful material for studies of the structure and biochemistry of individual chromosomes.

引用

页码：255 / 259

页数：5

共 16 条

[1] REPAIR OF X-RAY DAMAGE IN AGING WI-38 CELLS [J].

CLARKSON, JM ;

PAINTER, RB .

MUTATION RESEARCH, 1974, 23 (01) :107-112

[2] CONFORMATION OF DNA IN ALKALINE SUCROSE - SUBUNIT HYPOTHESIS IN MAMMALIAN-CELLS [J].

CLEAVER, JE .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1974, 59 (01) :92-99

[3] CHROMOSOME MEASUREMENT AND SORTING BY FLOW SYSTEMS [J].

GRAY, JW ;

CARRANO, AV ;

STEINMETZ, LL ;

VANDILLA, MA ;

MOORE, DH ;

MAYALL, BH ;

MENDELSOHN, ML .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1975, 72 (04) :1231-1234

[4]

HANSON CV, 1973, COLD SPRING HARB SYM, V38, P341

[5] EUKARYOTIC CHROMOSOME [J].

HEARST, JE ;

BOTCHAN, M .

ANNUAL REVIEW OF BIOCHEMISTRY, 1970, 39 :151-+

[6]

HOOPER DC, 1971, P SOC EXP BIOL MED, V136, P707

[7]

Horikawa M, 1977, Methods Cell Biol, V15, P97, DOI 10.1016/S0091-679X(08)60211-3

[8] SEPARATION OF ISOLATED CHINESE HAMSTER METAPHASE CHROMOSOMES INTO 3 SIZE-GROUPS [J].

MENDELSOHN, J ;

MOORE, DE ;

SALZMAN, NP .

JOURNAL OF MOLECULAR BIOLOGY, 1968, 32 (01) :101-+

[9] DIRECT APPROACH TO STRUCTURE OF EUKARYOTIC CHROMOSOMES [J].

SEDAT, J ;

MANUELIDIS, L .

COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY, 1977, 42 :331-350

[10] ISOLATION, FRACTIONATION AND BIOCHEMICAL ANALYSIS OF METAPHASE CHROMOSOMES OF MICROTUS-AGRESTIS [J].

SKINNER, LG ;

OCKEY, CH .

CHROMOSOMA, 1971, 35 (02) :125-&

← 1 2 →