THE PREPARATION OF HIGH-MOLECULAR-WEIGHT DNA FROM RICE AND ITS ANALYSIS BY PULSED-FIELD GEL-ELECTROPHORESIS

被引：8

作者：

HATANO, S ^{[1
]}

YAMAGUCHI, J ^{[1
]}

HIRAI, A ^{[1
]}

机构：

[1] NAGOYA UNIV,SCH AGR,GRAD DIV BIOCHEM REGULAT,NAGOYA 46401,JAPAN

来源：

PLANT SCIENCE | 1992年 / 83卷 / 01期

关键词：

ALPHA-AMYLASE; HIGH MOLECULAR WEIGHT DNA; ISOLATION OF NUCLEI; PULSED-FIELD GEL ELECTROPHORESIS; RDNA; RICE;

D O I：

10.1016/0168-9452(92)90062-Q

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：

We have isolated high molecular weight DNA of over 5.7 Mb in length from isolated rice germ nuclei. The high-molecular-weight DNA was digested with a number of restriction endonucleases. An ethidium-stained gel after pulsed-field electrophoresis revealed that few of these endonucleases have the ability to cleave rice chromosomal DNA to large fragments of over 200 kb in length, perhaps because of the lower level of methylation of nucleotides in the rice genome than in other genomes. Some restriction endonucleases that do not cut rDNA repeats produced fragments with arrays of rDNA repeats of more than 1.1 Mb, as estimated by Southern analysis. Therefore, it appears that the rice rDNA cluster is constructed with a 'physical' succession of more than 130 copies of 8-kb repeating units of rDNA. Genes for alpha-amylase were also detected by the Southern method. It indicates that our procedure is suitable for analysis of low-copy-number genes.

引用

页码：55 / 64

页数：10

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