+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Long distance RT-PCRs of human coronavirus 229E RNA



Long distance RT-PCRs of human coronavirus 229E RNA



Advances in Experimental Medicine and Biology 440: 269-273



The generation and cloning of cDNA fragments longer than 10 kb is often a difficult and time consuming task. In this study, we have analysed the conditions necessary of produce reverse transcripts longer than 10 kb that can be amplified by polymerase chain reaction. Thus, we isolated poly(A)-RNA from human coronavirus 229E infected MRC-5 cells and did reverse transcription using a sequence-specific primer. Subsequently, we amplified PCR products of varying length upstream of the primer position. Optimisation of the poly(A)-RNA preparation, the reverse transcription protocol and the polymerase chain reaction cycle conditions enabled us to successfully amplify regions of the human coronavirus 229E genome between 11.5 and 20.3 kb in length.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 046565933

Download citation: RISBibTeXText

PMID: 9782292


Related references

Coronavirus isolates SK and SD from multiple sclerosis patients are serologically related to murine coronaviruses A59 and JHM and human coronavirus OC43, but not to human coronavirus 229E. Journal of Virology 38(1): 231-238, 1981

Characterization of the human coronavirus 229E (HCV 229E) gene 1. Advances in Experimental Medicine and Biology 342: 75-79, 1993

Characteristics of a long term in vitro persistent infection with human coronavirus 229E. Advances in Experimental Medicine and Biology 142: 309-322, 1981

Characteristics of Australian human enteric coronavirus-like particles: comparison with human respiratory coronavirus 229E and duodenal brush border vesicles. Archives of Virology 97(3-4): 309-323, 1987

ADP-ribose-1"-phosphatase activities of the human coronavirus 229E and SARS coronavirus X domains. Advances in Experimental Medicine and Biology 581: 93-96, 2006

Antigenic studies on coronavirus 1. identification of the structural antigens of human coronavirus strain 229e. Canadian Journal of Microbiology 27(3): 334-342, 1981

Identification and characterization of a novel alpaca respiratory coronavirus most closely related to the human coronavirus 229E. Viruses 4(12): 3689-3700, 2012

The ADP-ribose-1''-monophosphatase domains of severe acute respiratory syndrome coronavirus and human coronavirus 229E mediate resistance to antiviral interferon responses. Journal of General Virology 92(Pt 8): 1899-1905, 2011

Comparative host gene transcription by microarray analysis early after infection of the Huh7 cell line by SARS coronavirus and human coronavirus 229E. Hong Kong Medical Journal 15(Suppl. 9): 23-26, 2009

Comparative host gene transcription by microarray analysis early after infection of the Huh7 cell line by severe acute respiratory syndrome coronavirus and human coronavirus 229E. Journal of Virology 79(10): 6180-6193, 2005

Distant relatives of severe acute respiratory syndrome coronavirus and close relatives of human coronavirus 229E in bats, Ghana. Emerging Infectious Diseases 15(9): 1377-1384, 2009

Neurotropism of human coronavirus 229E. Advances in Experimental Medicine and Biology 342: 339-346, 1993

Non structural proteins 8 and 9 of human coronavirus 229E. Advances in Experimental Medicine and Biology 581: 49-54, 2006

The genome of human coronavirus strain 229E. Journal of General Virology 39(3): 497-504, 1978