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Fig. 2 | Virology Journal

Fig. 2

From: Construction and characterization of an infectious cDNA clone of potato virus S developed from selected populations that survived genetic bottlenecks

Fig. 2

Schematic illustration of the construction of full-length cDNA clone of PVS-H00. Genome organization of PVS-H95 or PVS-H00 is shown at the top. The 3′ terminal half of PVS-H00 cDNA was amplified by PCR using PVS-37P and PVS-38 M as primers, and cloned to produce pPVS-37P38M. The 3′ terminal sequence downstream of the SnaBI restriction site in pPVS-H-37P38M was replaced with the 3′ terminal sequence containing a unique SpeI site immediately after the poly(A) tail of 66 adenine residues from pPVS-H-37P3ESpe2, which was a cDNA clone of PVS-H00. The 5′ terminal half of PVS-H00 cDNA was amplified by PCR using primers T7-PVS-H and PVS-37 M, and cloned to produce pPVS-T737 M. Finally, the 3′ terminal half of cDNA with the poly(A) tail and SpeI site from pPVS-H-37P38MSpe was connected with pPVS-T737 M via a unique EcoO65I site to produce the full-length cDNA clone pPVS-H-FL-β

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