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

Journal article

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 horse sickness virus (AHSV) have been cloned, terminally sequenced and expressed in vitro.

Authors: Vreede, F; Cloete, M; Napier, G; van Dijk, A; Viljoen, G

• Publication status: Published
• Journal: Journal of virological methods
• Volume: 72
• Issue: 2
• Pages: 243-247
• Publication date: 1998-06-01
• DOI: 10.1016/s0166-0934(98)00031-7
• EISSN: 1879-0984
• ISSN: 0166-0934
• Language: English
• Keywords: DNA, Complementary; Electrophoresis, Agar Gel; Horses; Genome, Viral; Gene Amplification; Animals; DNA Primers; Blotting, Northern; African horse sickness virus; RNA Viruses