发明名称 |
Covalent joining of DNA to RNA by vaccinia topoisomerase and uses thereof |
摘要 |
The present invention provides a method of covalently joining a DNA strand to an RNA strand comprising (a) forming a topoisomerase-DNA intermediate by incubating a DNA cleavage substrate comprising a topoisomerase cleavage site with a topoisomerase specific for that site, wherein the topoisomerase-DNA intermediate has one or more 5' single-strand tails; and (b) adding to the topoisomerase-DNA intermediate an acceptor RNA strand complementary to the 5' single-strand tail under conditions permitting a ligation of the covalently bound DNA strand of the topoisomerase-DNA intermediate to the RNA acceptor strand and dissociation of the topoisomerase, thereby covalently joining the DNA strand to the RNA strand. The present invention also provides a method of tagging a 5' end of an RNA molecule. The present invention further provides a DNA-RNA molecule which has been joined in vitro by the use of a topoisomerase. The present invention also provides a method of tagging a 5' end of an mRNA. The present invention provides a method of isolating and cloning full-lengh gene sequences using capped mRNA after subtraction of non-capped RNA.
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申请公布号 |
US2004058417(A1) |
申请公布日期 |
2004.03.25 |
申请号 |
US20030666486 |
申请日期 |
2003.09.19 |
申请人 |
SLOAN KETTERING INSTITUTE FOR CANCER RESEARCH |
发明人 |
SHUMAN STEWART;SEKIGUCHI JOANN;COMISKEY JOHN;FERNANDEZ JOSEPH;HOEFFLER JAMES;MARCIL ROBERT |
分类号 |
C12N15/09;C07H21/02;C07H21/04;C12N9/90;C12N15/10;C12P19/34;C12Q1/68;(IPC1-7):C12P19/34 |
主分类号 |
C12N15/09 |
代理机构 |
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地址 |
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