CLONING OF CHICKEN-EMBRYO TRANSFER-RNA GENES USING SINGLE STRANDED NUCLEOSOMAL DNA HIGHLY ENRICHED FOR TRANSFER-RNA COMPLEMENTARY SEQUENCES

被引：1
作者：
WITTIG, B
WITTIG, S
GRUNZ, H
机构：
[1] Institut für Molekularbiologie und Biochemie, Freie Universitàt Berlin, D-1000 Berlin 33
来源：
NUCLEIC ACIDS RESEARCH | 1979年 / 6卷 / 12期
关键词：
D O I：
10.1093/nar/6.12.3759
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：
DNA from chicken embryo nucleosome tetramers (about 760 base pairs in size) was enriched for tRNA genes by RPC-5 chromatography. The enriched DNA was hybridized with chicken embryo total tRNA and the hybridized DNA isolated utilizing a) avidin-biotin interaction, b) diazobenzyloxymethyl paper, and c) high temperature RPC-5 chromatography. The obtained single stranded DNA highly enriched for tRNA complementary sequences was hybridized with total DNA from nucleosome monomers ( 140-190 base pairs in size) and the excess of non hybridized monomer nucleosome DNA removed by Sepharose 4B chromatography. The hybrid molecules obtained were made fully double stranded by incubation with E.coli DNA polymerase I, DNA ligase, and exonuclease III. DNA was inserted into plasmid pBR322 by G-C joining procedure and the recombinant DNA used to transform the E.coli strain X1776. More than 70% of the transformants obtained hybridize to chicken embryo total tRNA. © 1979 Information Retrieval Limited.
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页码：3759 / 3772
页数：14
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