Construction of full-length cDNA clones of lettuce mosaic virus (LMV) and the effects of intron-insertion on their viability in Escherichia coli and on their infectivity to plants

1998 ◽  
Vol 143 (12) ◽  
pp. 2443-2451 ◽  
Author(s):  
S. J. Yang ◽  
F. Revers ◽  
S. Souche ◽  
H. Lot ◽  
O. Le Gall ◽  
...  
2021 ◽  
Vol 166 (5) ◽  
pp. 1427-1431
Author(s):  
Buyang Chen ◽  
Qi Lin ◽  
Yueyan Yin ◽  
Liangliang Jiang ◽  
Fang Wang ◽  
...  

1998 ◽  
Vol 36 (9) ◽  
pp. 647-656 ◽  
Author(s):  
Chye-Fong Liew ◽  
Chong-Jin Goh ◽  
Chiang-Shiong Loh ◽  
Saw-Hoon Lim

1998 ◽  
Vol 72 (1) ◽  
pp. 380-387 ◽  
Author(s):  
J. J. M. Meulenberg ◽  
J. N. A. Bos-de Ruijter ◽  
R. van de Graaf ◽  
G. Wensvoort ◽  
R. J. M. Moormann

ABSTRACT The 5′-terminal end of the genomic RNA of the Lelystad virus isolate (LV) of porcine reproductive and respiratory syndrome virus was determined. To construct full-length cDNA clones, the 5′-terminal sequence was ligated to cDNA clones covering the complete genome of LV. When RNA that was transcribed in vitro from these full-length cDNA clones was transfected into BHK-21 cells, infectious LV was produced and secreted. The virus was rescued by passage to porcine alveolar lung macrophages or CL2621 cells. When infectious transcripts were transfected to porcine alveolar lung macrophages or CL2621 cells, no infectious virus was produced due to the poor transfection efficiency of these cells. The growth properties of the viruses produced by BHK-21 cells transfected with infectious transcripts of LV cDNA resembled the growth properties of the parental virus from which the cDNA was derived. Two nucleotide changes leading to a unique PacI restriction site directly downstream of the ORF7 gene were introduced in the genome-length cDNA clone. The virus recovered from this mutated cDNA clone retained the PacI site, which confirmed the de novo generation of infectious LV from cloned cDNA. These results indicate that the infectious clone of LV enables us to mutagenize the viral genome at specific sites and that it will therefore be useful for detailed molecular characterization of the virus, as well as for the development of a safe and effective live vaccine for use in pigs.


BioTechniques ◽  
2004 ◽  
Vol 36 (4) ◽  
pp. 690-700 ◽  
Author(s):  
Jia Qian Wu ◽  
Angela M. Garcia ◽  
Steven Hulyk ◽  
Anna Sneed ◽  
Carla Kowis ◽  
...  

2007 ◽  
Vol 153 (1) ◽  
pp. 181-185 ◽  
Author(s):  
D. Stephan ◽  
I. Moeller ◽  
A. Skoracka ◽  
F. Ehrig ◽  
E. Maiss

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