Sequence-independent amplification and cloning of large dsRNA virus genome segments by poly(dA)-oligonucleotide ligation

被引：24

作者：

Vreede, FT ^{[1
]}

Cloete, M ^{[1
]}

Napier, GB ^{[1
]}

van Dijk, AA ^{[1
]}

Viljoen, GJ ^{[1
]}

机构：

[1] Onderstepoort Vet Inst, ZA-0110 Onderstepoort, South Africa

来源：

JOURNAL OF VIROLOGICAL METHODS | 1998年 / 72卷 / 02期

关键词：

dsRNA viruses; African horse sickness virus (AHSV); single primer amplification;

D O I：

10.1016/S0166-0934(98)00031-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A strategy was developed for sequence-independent synthesis and amplification of full-length cDNA of 3-4 kb genes of dsRNA viruses. The method of single primer amplification (Lambden et al., 1992) was adapted by the inclusion of a 3' poly(A) tail to an oligonucleotide ligated to dsRNA genome segments as a template for oligo(dT)-primed cDNA synthesis. Full-length copies of the largest genome segments, 1 (4 kb) and 2 (3 kb), of African horst sickness virus (AHSV) have been cloned, terminally sequenced and expressed in vitro. (C) 1998 Elsevier Science B.V. All rights reserved.

引用

页码：243 / 247

页数：5

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