Genic amplification of the entire coding region of the HEF RNA segment of influenza C virus.
In order to provide an easy and powerful analysis of influenza C viral HEF RNA segment of a recent strain, a combination of reverse transcription and the polymerase chain reaction was used. We amplified the entire coding region of the HEF gene of a laboratory strain of virus called C/Johannesburg/1/66, widely used for binding and esterase activity studies as well as that of a strain isolated in 1991 (C/Paris/145/91) from a patient suffering from severe flu syndrome. The sequences we amplified were about 2 kilobases long. In this work, we show that the forward 'universal primer' Uni1, which has been used for influenza A and B viruses cDNA syntheses can also be used for influenza C virus. The PCR primers were designed to contain restriction sites to make the PCR products ready to be used for further purposes. A restriction analysis of the PCR products combined with analyses of all the human influenza C virus HEF gene sequences published so far permitted the design of sets of oligonucleotides which can prime PCR on cDNA of unknown influenza C virus for cloning.
Manuguerra
J C
JC
Institut Pasteur, Unité d'Ecologie Virale, Paris, France.
Hannoun
C
C
Nicolson
C
C
Robertson
J S
JS
D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics; D005784:Gene Amplification; D005814:Genes, Viral; D006801:Humans; D007251:Influenza, Human / Q000382:microbiology; D009982:Gammainfluenzavirus / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics*; D014773:Virology / Q000379:methods*
D004279:DNA, Viral; D012367:RNA, Viral; D004262:DNA Restriction Enzymes