Nucleic Acids Research, 1977, Vol. 4, No. 7 2145-2160
© 1977
Articles |
Reverse transcription of phage RNA and its fragment directed by synthetic heteropolymeric primers
Institute of Molecular Biology Acad. of Sci., Moscow V-312 Moscow State University Acad. of Sci of Latvian SSR, Riga, USSR Institute of Organic Synthesis Acad. of Sci of Latvian SSR, Riga, USSR
+To whom all correspondence should be addresseds
Received March 18, 1977. DNA synthesis catalysed by RNA-directed DNA-polymerase (reverse tranacriptase)was found to proceed on the ERA template of an MS2 phage in the presence of heteropolymeric synthetic octa- and nonadeoxyribonucleotide primers complementary to the intercistronic region (coat protein binding site) and the region of the coat protein cistron, respectively. The product of synthesis consists of discrete DNA fractions of different length, including transcripts longer than 1,000 nucleotides. The coat protein inhibits DNA synthesis if it is initiated at its binding site, but has no effect on DNA synthesis initiated at the coat protein cistron. It has been suggested that, in this system, the initiation of DNA synthesis by synthetic primers is topographically specific. The MS2 coat protein binding site (an RNA fragment of 59 nucleotides) serves as a template for polydeoxyribonucleotide synthesis in the presence of octanucleotide primer and reverse transcriptase. The product of synthesis is homogenous and its length corresponds to the length of the template. The effective and complete copying of the fragment having a distinct secondary structure proves that the secondary structure does not interfere, in principle, with RNA being a template in the system or reverse transcription.